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T Papp

Publications and source records attributed to T Papp.

At least 19 recordsLinked to original sources

Genetic variability of the postharvest pathogen Gilbertella persicaria: identification of randomly amplified polymorphic DNA (RAPD) markers correlating with (+) and (-) mating types.

Random amplified polymorphic DNA (RAPD) and isoenzyme polymorphisms among 16 isolates of the postharvest pathogen Gilbertella persicaria were examined. Six different 10-bp primers were used to determine the extent of intraspecific genetic variability. Nine composite amplification types were identified. RAPD markers were obtained which correlated with the mating types of the G. persicaria isolates. The variability of the isoenzyme patterns was very low and no correlation was found between the isoenzyme markers and the mating abilities. When 80 single carbon substrates were tested in utilization assays, most of them were utilized uniformly by the 16 G. persicaria strains. However, some compounds elicited differences between the isolates representing the two mating types. Beta-alanine (0.2%) has little effect on the germination of the sporangiospores of the (+) isolates, but inhibited the germination of (-) sporangiospores. Glycerol-1-monoacetate supported the growth of both mating types, but at concentrations higher than 4% this was accompanied with a compact (colonial) growth for plus mating type isolates only.

Carbon↗

Presence of double-stranded RNA and virus-like particles in Rhizopus isolates.

Fungal isolates belonging to four Rhizopus species were screened for the presence of double-stranded RNA (dsRNA) molecules. Five (two R. stolonifer, two R. microsporus, and one R. oryzae) of the 27 isolates examined harboured such genetic elements. Electrophoresis of the nucleic acids revealed five RNA patterns, with 1-5 discrete dsRNA bands. The molecular sizes corresponding to these bands were 2.2-14.8 kb. Gel electrophoresis of purified virus-like particles (VLPs) indicated only one capsid of similar size in all virus-harbouring strains; when investigated by electron microscopy, they were found to be polyhedral VLPs 40 nm in diameter. In one of the R. microsporus isolates an uncapsidated large dsRNA segment (14.8 kb) was observed. No phenotypic differences were observed between uninfected and virus-harbouring Rhizopus isolates.

RNA, Double-Stranded↗

Variation of isoenzyme and RAPD patterns in Candida albicans morphological mutants with altered colony ultrastructure.

Molecular typing methods were applied to characterize four stable morphological mutants [1] isolated from a UV-induced unstable mutant colony of Candida albicans. The wild-type strain (ATCC 64385), the intermediate unstable mutant and its four morphologically altered derivatives revealed the same electrophoretic karyotypes. Of the five isoenzymes tested (catalase, malate dehydrogenase, glutamate dehydrogenase, acid phosphatase and 3-glucosidase), glutamate dehydrogenase displayed a different enzyme pattern (with an extra band of lower mobility) in the morphological mutants. In contrast, the random amplification DNA polymorphism patterns of the mutant strains differed in all cases from that of the parental strain. Different primers revealed various degrees of DNA polymorphism; one of them (OPC-8) proved to be useful for differentiation between all examined strains. Differences in genetic alterations between spontaneous and induced mutants, and the applicability of different molecular markers to analyse the consequences of induced mutagenesis in C. albicans are discussed.

Candida albicans↗

Variability of isozyme and rapd markers among isolates of Mucor genevensis.

Mucor genevensis is a dimorphic and homothallic fungal species (Zygomycetes). Ten M. genevensis strains, each strain of the recently described new homothallic species (M. meguroense and M. hachijyoensis) and strains of M. hiemalis and M. piriformis (as outgroups for numerical analysis) were investigated. Five different enzyme systems (CAT, GDH, G6D, MDH and SOD) and five 10-bp random primers were used in isoenzyme and random amplified polymorphic DNA analyses, respectively. The data from these studies were subjected to numerical analyses. Substantial intraspecific variability was detected in M. genevensis with both of the methods applied. Though both the M. meguroense strain and the M. hachijyoensis strain revealed characteristic differences, they grouped closer to the homothallic M. genevensis than to the heterothallic M. piriformis and M. hiemalis strains.

Genetic Markers↗

Mutational analysis of N-ras, p53, p16INK4a, p14ARF and CDK4 genes in primary human malignant mesotheliomas.

Nineteen specimens from primary human malignant mesotheliomas obtained from 19 patients were screened for activating point mutations in the oncogenes N-ras and CDK4 by combined RFLP-PCR/SSCP analysis. In addition, all tumours were screened for deletions and point mutations in the tumour suppressor genes p53, p16INK4a (CDKN2A) and p14ARF (exon-1beta) by combined multiplex-PCR/SSCP analysis. No mutations were found in N-ras, p53 and CDK4. Three tumours displayed homozygous deletion (co-deletion of exons 1, 2 and 3) of p16INK4a. One of them displayed additional homozygous deletion of p14ARF (exon-1beta). Two silent point mutations and 2 polymorphisms were found in p16INK4a in 3 tumours. Our preliminary data indicate that disarrangement of the Rb1 pathway may be involved in mesothelioma formation.

Adult↗

Intraspecific variation in two species of Rhizomucor assessed by random amplified polymorphic DNA analysis.

Twenty-three Rhizomucor isolates were characterized by random amplified polymorphic DNA-PCR (RAPD-PCR) with 10-bp oligonucleotide primers. These data were used for numerical analyses to obtain information on the intraspecific genetic polymorphism of Rhizomucor species. The genetic variability in Rhizomucor pusillus and Rhizomucor miehei isolates was found to differ; the latter revealed less intraspecific polymorphism. The different levels of genotypic diversity suggest a correlation with the different forms of mating behaviour of these species. Rhizomucor tauricus displayed amplification patterns similar to those of the investigated R. pusillus strains, reinforcing the assumption that R. tauricus does not represent a separate species. Characteristic RAPD markers allowing PCR-based species identification of Rhizomucor isolates were determined.

Animals↗

The mitochondrial genome of Mucor piriformis.

DNA was purified from the isolated mitochondria of a Mucor piriformis wild-type strain (NRRL 26211). A circular restriction map of the mitochondrial DNA was established on the basis of single and double digests with several restriction endonucleases. The average mitochondrial DNA size calculated from these data was 33.53 kbp; this is in good agreement with the contour length size (33.62 kbp) of the open circular molecules detected by electron microscopy. Heterologous hybridizations with cloned Aspergillus nidulans mitochondrial genes were used to locate some coding regions on the map.

DNA, Fungal↗

Mutational analysis of the N-ras, p53, p16INK4a, CDK4, and MC1R genes in human congenital melanocytic naevi.

Eighteen human congenital melanocytic naevi (CMN) from 17 patients were screened for activating point mutations in the oncogenes N-ras and CDK4 and for sequence variants in the MC1R gene by combined RFLP-PCR/SSCP analysis. In addition, all lesions were screened for deletions and point mutations in the tumour suppressor genes p53 and p16INK4a (CDKN2A) by combined multiplex PCR/SSCP analysis. Positive screening data were specified by sequencing of the corresponding PCR product. Activating point mutations in the N-ras gene (nine CAA (Gln) to AAA (Lys) transversions and one CAA (Gln) to CGA (Arg) transition at codon 61) were detected at high frequency (56%). Furthermore, three missense mutations (V92M) and two silent mutations (CGA (Arg) to CGG (Arg), codon 213, exon 6) were found in the MC1R and p53 genes, respectively. No mutations were found in p16 or CDK4. The activated N-ras oncogene, which is also found in human cutaneous melanomas, may constitute a potential risk factor for melanoma formation within CMN.

Adolescent↗

Detection of double-stranded RNA molecules and virus-like particles in different Mucor species.

The presence of double-stranded RNA elements was examined in 123 strains representing 18 Mucor species. These genetic elements were found to be present in 6 strains: 1 M. aligarensis, 1 M. hiemalis, 2 M. corticolus, 1 M. mucedo and 1 M. ramannianus. Electrophoretic separation of the nucleic acids revealed 4 different RNA patterns, with 1 to 5 discrete dsRNA bands. The molecular weights corresponding to these bands were 1.42-4.15 x 10(6) D. Using electronmicroscopy, for the first time the presence of virus like particles in Mucor species has been revealed.

Electrophoresis, Agar Gel↗

Differentiation of Rhizomucor species by carbon source utilization and isoenzyme analysis.

Nineteen Rhizomucor miehei and Rhizomucor pusillus isolates were assayed for their ability to utilize 87 various substrates as a single carbon source. Besides a difference in sucrose utilization, distinctive differences were found in the utilization of glycine, phenylalanine, and beta-alanine. Five isoenzyme systems also proved useful for the determination of markers of distinctive value at a species level. Data were used to obtain information about the genetic polymorphism of these species: a high degree of variability was found among the R. pusillus isolates, whereas the group of R. miehei isolates was more homogeneous genetically.

Animals↗

Detection of hyperdiploidy and chromosome breakage affecting the 1 (1cen-q12) region in lentigo malignant melanoma (LMM), superficial spreading melanoma (SSM) and congenital nevus (CN) cells in vitro by the multicolor FISH technique.

The centric/pericentric region of chromosome 1 (cen-q 2) of human melanoma cells of different stages of carcinogenicity (superficial spreading melanoma (SSM), lentigo malignant melanoma (LMM)) and premalignant precursor lesions (congenital nevus (CN)) were investigated by fluorescence in situ hybridization (FISH) with tandem DNA probes. The pericentric heterochromatin region 1(q12) is large and highly prone to breakage in contrast to the adjacent centromeric region which is much smaller and less prone to such events. All samples of melanoma cells were obtained from patients and cultivated in vitro. LMM cells showed the highest number of breakage events within the 1q12 region (90% of cells). The number of hyperdiploid cells was not increased in comparison to CN cells. In contrast to LMM cells, SSM cells showed a significant increased number of hyperdiploid cells which were mainly tetrasomic for chromosome 1 (P < or = 0.05). The number of chromosome breaks was not significantly increased in this type of melanoma cells. The spontaneous rates of chromosomal breakage and hyperdiploidy is relatively low in CN cells (1.5-2.5% and 3.2-5.8%, respectively) but these frequencies also differ between CN samples from different patients. These results show that the multicolor FISH technique represents a fast and reliable detection method, distinguishing structural and numerical chromosomal alterations in interphase nuclei. This technique is useful as a histological marker to differentiate between specific tumor subtypes and to investigate the relationship between genomic instability and clinopathological parameters (tumor grading and staging).

Aneuploidy↗

Significant antenatal factors in the development of lumbar spinal stenosis.

STUDY DESIGN: Adverse factors during pregnancy may permanently stunt the growth of the spinal canal. Subsequently, even in an optimal environment the canal cannot catch up in growth with the trunk and long bones because of its early maturation. The degree of retardation in canal size depends on the severity and timing of the adverse effect. The catch-up growth of the long bones mask the narrow canal, because the latter does not have growth potential, resulting in an adult of sufficient height and good proportions, but with a canal at risk for stenosis. OBJECTIVES: To investigate the influence of the antenatal environment on the growth of the lumber spinal canal. SUMMARY OF BACKGROUND DATA: To date, little is known about the effects of an adverse environment on the growth of the spinal canal, and no data have been reported on antenatal influencing factors. METHODS: Lumbar magnetic resonance imaging scans from 58 patients were examined. Dimensions of the central spinal canals were measured by computerized image analysis and compared with the subjects' obstetric data from their mothers' pregnancies. RESULTS: The L3 canal was found to be the most sensitive to the influence of the examined factors. Gestational age was the most significant factor; if short, it resulted in small adult canal. Small placental weight, greater maternal age, primiparity, low socioeconomic class, and low birth weight were also found to be significant in affecting the growth of the canal. CONCLUSIONS: An adverse antenatal environment does have a permanent, retarding effect on the growth of the lumbar spinal canal.

Adolescent↗

An in vitro study of the biomechanical effects of flexible stabilization on the lumbar spine.

STUDY DESIGN: Lumbar motion segments were tested in vitro to examine biomechanical changes after posterior fixation by a flexible device. OBJECTIVES: To assess changes in load distribution and conformation of vertebral structures after a flexible stabilization. This should provide the foundations for a scientific understanding of the immediate effects of this surgical procedure. METHODS: Hooks were placed over the proximal spinous process and the distal laminas of a motion segment and connected by a polyester braid. Tension applied to the braid then generated a compression of the posterior elements. The force between the articular facets, the displacement of the posterior anulus fibrosus of the intervertebral disc, and the change in the relative position of the adjacent vertebrae were measured as the applied tension was increased. RESULTS: Facet joint force, disc bulge, and vertebral angulation increased with applied tension until a position of "locking" was achieved, apparently when the bony margin of the superior half of the facet joint contacted the inferior pars interarticularis. A tension of between 50 to 100 N in the braid was required for this. Facet joint force was less than 40% of this, and disc bulge was only 0.15 mm. The extension of the motion segment was between 2 degrees and 8 degrees. CONCLUSIONS: The results suggest that if such a system is applied surgically, stabilization is produced by compaction of the bony margins of the facet joints. Only a relatively small proportion of the posteriorly applied load is carried by the facet joints themselves, and little angulatory change is expected with minimal disc bulge.

Aged↗

DNA amplification polymorphisms of Mucor piriformis.

Mucor piriformis can cause postharvest decay in various fruits and vegetables stored at low temperatures. Thirty isolates of this fungus, collected from infected fruit, were subjected to random amplified polymorphic DNA (RAPD) analysis. Seven different 10-bp primers were used to determine the type and extent of intraspecific genetic polymorphisms. Nineteen composite amplification types were identified, indicating a higher degree of variability than found in previous isoenzyme studies. Numerical analysis with the UPGMA technique revealed three clusters, which correlated with the mating competency of the isolates or their place of origin. These results demonstrate that RAPD analysis can identify isolates and subspecific populations of M. piriformis.

DNA Primers↗

Lack of p53 mutations and loss of heterozygosity in non-cultured human melanocytic lesions.

In this study we analysed snap-frozen surgical resections of 16 superficial spreading melanomas, 13 nodular malignant melanomas, 2 lentigo maligna melanomas, 1 dysplastic nevus, 1 congenital nevus and 5 normal nevi from 38 patients for point mutations in the human p53 gene at exons 5-8 by polymerase chain reaction/single-strand conformation polymorphism as well as for loss of heterozygosity of p53 by restriction-fragment-length polymorphism/polymerase chain reaction in order to determine whether p53 aberrations are associated with melanoma subtypes. In addition, we analysed six melanoma cell lines for point mutations in p53. Our results revealed the absence of point mutations and loss of heterozygosity in all fresh resected lesions. However, a TAC (Tyr) to TGC (Cys) transition at codon 163 in exon 5 was found in one cell line.

Base Sequence↗

Analysis of ras mutations in human melanocytic lesions: activation of the ras gene seems to be associated with the nodular type of human malignant melanoma.

We have analyzed the Ha-ras, Ki-ras and N-ras gene for point mutations at codons 12, 13 and 61 via restriction fragment length polymorphism/polymerase chain reaction analysis and subsequent direct sequencing in non-cultured fresh-frozen tissues of 16 superficial spreading melanomas (SSM), 13 nodular malignant melanomas (NMM), 2 lentigo malignant melanomas (LMM), 1 dysplastic nevus, 1 congenital nevus and 5 normal nevi from 38 patients. Mutations were found in 4 melanoma samples, all belonging to the nodular malignant type. Three of them were mutated in N-ras and one in the Ha-ras gene. Mutation in N-ras was also detected in the congenital nevus. All mutations were exclusively located at the first two base pairs of codon 61. No Ki-ras mutation was detected in any lesion. No mutation could be found in SSM and LMM in addition to dysplastic and normal nevi. The frequency of ras mutation in NMM was 31%, whereas in SSM it was 0%. Our study suggests (a) an association between ras mutations (mainly N-ras) and the NMM as a subgroup of human melanoma; (b) that activation of Ki-ras is not involved in the pathogenesis of melanoma. The role of UV radiation in point mutations of ras genes in human melanoma is discussed.

Base Sequence↗