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Restriction-fragment-length polymorphism analysis of small-subunit rRNA genes of Blastocystis hominis isolates from geographically diverse human hosts.

Genomic diversity among 14 isolates of Blastocystis hominis from 4 different geographic locations was examined by small-subunit rRNA (ssu rRNA) restriction-fragment-length polymorphisms (RFLP) using 5 different restriction endonucleases. On the basis of the observed RFLP patterns among the isolates, a total of 12 genotypes were identified, with 7 isolates exhibiting mixed RFLP genotypes. There was no correlation between B. hominis geographic origin and RFLP banding pattern or genotype.

Animals↗

Relationship between primary Gleason pattern on needle biopsy and clinicopathologic outcomes among men with Gleason score 7 adenocarcinoma of the prostate.

OBJECTIVES: To examine the relationship among needle biopsy primary grade, prostatectomy grade, and postprostatectomy biochemical recurrence among men with Gleason score 7 disease. METHODS: We identified 320 men with Gleason score 7 tumors on prostate biopsy treated with radical prostatectomy between 1991 and 2001 by a single surgeon. None of these patients had received neoadjuvant or adjuvant hormonal therapy or radiotherapy. The chi-square test and Kaplan-Meier method were used to evaluate the correlation among biopsy Gleason score, prostatectomy Gleason score, and biochemical recurrence. RESULTS: A total of 252 (79%) and 68 (21%) men had primary Gleason pattern 3 and 4 identified on needle biopsy, respectively. Of the patients with Gleason pattern 3 + 4 tumors on biopsy, 24% were upgraded to primary pattern 4 or more on final pathologic analysis. Of the patients with Gleason pattern 4 + 3 tumors on biopsy, 47% were downgraded to primary pattern 3 or less on final pathologic analysis. The actuarial risk of biochemical prostate-specific antigen recurrence was significantly lower among patients with Gleason pattern 4 + 3 on biopsy, if the prostatectomy Gleason score was downgraded to 3 + 4 or less (P = 0.03). CONCLUSIONS: Approximately 47% of men with a diagnosis of Gleason pattern 4 + 3 on needle biopsy are downgraded at radical prostatectomy and will have biochemical prostate-specific antigen recurrence-free outcomes similar to patients originally diagnosed with Gleason pattern 3 + 4 adenocarcinoma. This group of patients may benefit from definitive treatment such as radical prostatectomy for management of their disease.

Adenocarcinoma↗

Independent clonal origins of distinct tumor foci in multifocal papillary thyroid carcinoma.

BACKGROUND: Papillary thyroid carcinoma is frequently multifocal. We investigated whether noncontiguous tumor foci arise from intraglandular metastases from a single primary tumor or originate as unrelated clones derived from independent precursors. METHODS: Using a polymerase-chain-reaction assay involving the human androgen receptor gene (HUMARA), we analyzed the patterns of X-chromosome inactivation of multiple distinct foci of well-differentiated multifocal papillary thyroid cancer from 17 women. RESULTS: Multiple thyroid tumor foci from 10 of 17 patients yielded DNA of adequate quality and were heterozygous for the HUMARA polymorphism and hence suitable for analysis. A single X chromosome was inactivated in each focus, consistent with its monoclonality. When the specific monoclonal configurations of each patient's discrete tumor foci were compared, discordant patterns indicative of independent origins were observed among the tumors from five patients; results in the remaining five were consistent with either a shared or independent clonal origin. CONCLUSIONS: Individual tumor foci in patients with multifocal papillary thyroid cancer often arise as independent tumors.

Carcinoma, Papillary↗

Regeneration of normal terminal innervation patterns by central noradrenergic neurons after 5,7-dihydroxytryptamine-induced axotomy in the adult rat.

The regeneration of central adrenergic axons has been followed between 5 days and 18 months after 5,7-dihydroxytryptamine(5,7-DHT)-induced axotomy in the adult rat, using fluorescence histochemistry in combination with noradrenaline (NA) determinations and [3H]NA uptake measurements. The axonal and terminal degeneration caused by the 5,7-DHT treatment (150 micrograms intraventricularly) was, by 1--2 weeks after injection, accompanied by a 70% reduction of NA in the forebrain and 30% reduction in the brain stem, and by 43--85% reductions in the [3H]NA uptake capacity in various regions of the brain and spinal cord. Signs of sprouting of the drug-lesioned axons were evident along the terminal axon segments at 5 days after treatment. The sprouts increased rapidly in length and number during the subsequent weeks and by 2--6 months after injection new NA terminal systems of relatively normal density and distribution had been re-established in many initially denervated regions. In parallel there was a recovery of endogenous NA and [3H]NA uptake to the pre-injection levels in the brain, and to levels 50--75% of normal in the cervical and thoracic spinal cord. Four successive phases of the regeneration process are distinguished: (1) primary sprouting from the lesioned NA axon stumps, occurring within the first week after treatment; (2) seemingly random growth and proliferation of the newly formed sprouts during the second and third weeks; (3) directed, forward growth of some of the sprouts leading to a partial restoration of the original fibre paths, branching patterns and terminal networks within 3--6 months; (4) a concomitant removal of at least part of the abnormally directed sprouts. Although the original fibre architecture was quite accurately restored in many areas the regeneration was not always correct. Hyperinnervation patterns and abnormal terminal arrangements were often formed, and in the spinal cord the down-growth of the regenerating axons occurred predominantly along a route that is inconspicuous in the normal rat. It is concluded that at least certain types of central neurons regenerate very efficiently provided the conditions are favourable, and that under such conditions axonal regeneration in the mammalian CNS is subjected to regulatory mechanisms that can be very precise. The results provide evidence that the adult mammalian CNS possesses mechanisms for axonal guidance which allow the accurate regeneration of lesioned axonal tracts and branching patterns, as well as mechanisms of recognition making possible the re-establishment of the original terminal connections.

5,7-Dihydroxytryptamine↗

Functional modification of acrylic resin monoplane occlusions.

Patients with acrylic-resin monoplane occlusions were recalled over a 6-month to 6-year period. The intention was to obtain casts of the dentures in order to study the anteroposterior wear pattern for classifiable variations from the original monoplane occlusion. The lateral component of the wear pattern was not examined during this study. The project revealed wear patterns that were classifiable in terms of the recognized prosthetic occlusal plane forms--namely, the Monson curve, a reverse curve, or a persistence of the original monoplane. Right and left aspects of the maxillary occlusal plane were examined as separate entities and lateral asymmetry was evident in most patients. No attempt was made to quantify the extent of wear as in the studies of Harrison.

Acrylic Resins↗

Patterns arising from the interaction between scalar and vectorial instabilities in two-photon resonant Kerr cavities.

We study pattern formation associated with the polarization degree of freedom of the electric field amplitude in a mean field model describing a nonlinear Kerr medium close to a two-photon resonance, placed inside a ring cavity with flat mirrors and driven by a coherent x-polarized plane-wave field. In the self-focusing case, for negative detunings the pattern arises naturally from a codimension two bifurcation. For a critical value of the field intensity there are two wave numbers that become unstable simultaneously, corresponding to two Turing-like instabilities. Considered alone, one of the instabilities would originate a linearly polarized hexagonal pattern whereas the other instability is of pure vectorial origin and would give rise to an elliptically polarized stripe pattern. We show that the competition between the two wave numbers can originate different structures, the detuning being a natural selection parameter.

Journal Article↗

Successful isolation, cultivation and partial characterization of naturally occurring ovine squamous cell carcinoma.

Seventeen epithelial cell lines have been successfully established from naturally occurring ovine squamous cell carcinomata. Culture establishment was most successful when tumor tissue was directly explanted rather than treated enzymatically. Success in establishing cultures also appeared to be related to the site on the body from which the tumor biopsy was taken, with tumors derived from the nose being most readily cultured. Several of the cell lines were successfully transplanted to nude mice where the growth patterns observed in the original host, i.e. expansive or invasive, were maintained. All cell lines assumed one of two distinct morphological types; however, no association could be seen between morphology and pattern of in vivo growth.

Animals↗

Use of non-neutralizing monoclonal antibodies in an ELISA for intratypic differentiation of 28 echovirus type 25 clinical isolates.

Three non-neutralizing monoclonal antibodies were produced and selected against echovirus type 25 JV-4 prototype strain. They were used in an ELISA to investigate the intratypic differentiation of 28 wild isolates. Clinical isolates fell into seven different groups according to their reactivity patterns in ELISA. Two of the non-neutralizing monoclonal antibodies, 9E4 and 6D3, were highly specific, while the third, 6C9, may recognize an epitope common to other types of echoviruses. In contrast, mouse polyclonal antiserum exhibited large cross-reactivities among echovirus serotypes. The reactivity patterns and the geographical origin of the isolates were generally not correlated and, in the same area, four major antigenic variants sometimes coexisted, especially in the south of France. Moreover, reactivity patterns found with ELISA were hardly ever correlated with those observed in a previous study when neutralization tests were used. These results again underline the non-correlation between structure and biological function in the Picornavirus family.

Animals↗

The evolution of trypanosomes infecting humans and primates.

Based on phylogenetic analysis of 18S rRNA sequences and clade taxon composition, this paper adopts a biogeographical approach to understanding the evolutionary relationships of the human and primate infective trypanosomes, Trypanosoma cruzi, T. brucei, T. rangeli and T. cyclops. Results indicate that these parasites have divergent origins and fundamentally different patterns of evolution. T. cruzi is placed in a clade with T. rangeli and trypanosomes specific to bats and a kangaroo. The predominantly South American and Australian origins of parasites within this clade suggest an ancient southern super-continent origin for ancestral T. cruzi, possibly in marsupials. T. brucei clusters exclusively with mammalian, salivarian trypanosomes of African origin, suggesting an evolutionary history confined to Africa, while T. cyclops, from an Asian primate appears to have evolved separately and is placed in a clade with T. (Megatrypanum) species. Relating clade taxon composition to palaeogeographic evidence, the divergence of T. brucei and T. cruzi can be dated to the mid-Cretaceous, around 100 million years before present, following the separation of Africa, South America and Euramerica. Such an estimate of divergence time is considerably more recent than those of most previous studies based on molecular clock methods. Perhaps significantly, Salivarian trypanosomes appear, from these data, to be evolving several times faster than Schizotrypanum species, a factor which may have contributed to previous anomalous estimates of divergence times.

Animals↗

Hepatitis A immunity in the Swedish population. A study of the prevalence of markers in the Swedish population.

After a 20-year interval, the prevalence of seroimmunity to Hepatitis A (HA) was again investigated in a statistical sample of the adult Swedish population. Sera from 3382 of the 4800 originally selected persons were tested. The prevalence of antibodies to HA had not changed since the 1960s when only the Scandinavian population was considered. In the oldest population born at the beginning of this century, the presence of antibodies amounted to 69%. It gradually declined to 6% in those born in the 1940s. In the population born after 1950, the percentage of seropositive individuals was only 2%. A slightly higher prevalence was seen in the big cities, compared with the rural areas (13% vs 9%). Persons of non-Scandinavian origin showed a different pattern. Those from other European countries showed a prevalence of about 70% in all the age-groups investigated. Among the young adults of Arabic or Asiatic origin, the figure was > 90%. The conclusion is that the native Swedish population has a low natural exposure to HA, which has not changed during the last 20 years. Prophylaxis before going to countries where the disease is endemic is strongly recommended.

Adolescent↗

Migration of craniofacial periosteum in growing guinea-pigs.

The use of a carbon particle tattoo provided stable periosteal markers and a means of recording periosteal movement both anteroposteriorly and transversely during growth in guinea-pigs. In general, the periosteum migrated toward the cranial sutures. The radial pattern, demonstrated on the frontal bones and indicated on the nasal and parietal bones, showed that a periosteal envelope is identifiable with each bone. The area of origin of this centrifugal pattern of migration coincided with the ossification centre of that bone. Trabeculae and vascular canals tended to point in the direction of periosteal migration. The anteroposterior periosteal migration was proportional to but less than the anteroposterior growth of the craniofacial bones studied. The absolute medial periosteal marker migration, similar on the frontal and parietal bones, was directed in the opposite direction from the slight transverse bone growth. This may indicate a biophysical response to tension on the fibrous periosteum from the principally anteroposterior craniofacial growth. The amount of growth at the frontonasal, coronal and midline sutures diminished from anterior to posterior. The observed histological and morphological sutural characteristics concurred with these growth changes which correlate with the decreased cranial and increased facial growth during the age period studied. Although the fibrous periosteum is continuous over the separate membranous bones its behaviour appears to be intimately related to the growth of the craniofacial bones which it covers. The blending of the fibrous periosteum within the transverse sutures may prevent significant migration across craniofacial bones. It seems that the observed migratory patterns result from a close association of the fibrous periosteum with each growing craniofacial bone. The relationship of the fibrous periosteum to the growth of the craniofacial bones is a question requiring further investigation.

Animals↗

Localization of temporal lobe foci by ictal EEG patterns.

Identifying patients whose complex partial seizures originate in temporal neocortex rather than in hippocampus is important because such patients have less favorable outcomes with standard anteromesial temporal resections. We reviewed scalp-recorded ictal EEGs of 93 epilepsy surgery candidates who either underwent intracranial EEG monitoring (n = 58) or who were referred directly for temporal lobectomy (n = 35). We definded seven patterns of early seizure discharges, grouped patients according to their seizure pattern, and correlated these with the site of seizure onset determined by intracranial EEG. Categorization by seizure pattern was also compared with brain magnetic resonance imaging (MRI) findings intracarotid amobarbital (Wada) testing. An initial, regular 5- to 9- Hz inferotemporal rhythm (type 1A) was most specific for hippocampal-onset seizures. Less commonly, a similar vertex/parasagittal positive rhythm (type 1B) or a combination of types 1B and 1A rhythms (type 1C) was recorded. Seizures originating in temporal neocortex were most often associated with irregular, polymorphic, 2- to 5-Hz lateralized activity (type 2A). This pattern was commonly followed by a type 1A theta rhythm (type 2B) or was preceded by repetitive, sometimes periodic, sharp waves (type 2C). Seizures without a clear lateralized EEG discharge (type 3) were most commonly of temporal neocortical origin. These associations between type of seizure pattern and probable site of cerebral origin were statistically significant. MRI and Wada testing did not have as much specificity as ictal patterns in differentiating among seizure origins. We conclude that the initial pattern of ictal discharge on scalp EEG can assist in distinguishing seizures of temporal neocortical onset from those of hippocampal onset. This information can be used to identify patients for invasive monitoring.

Amobarbital↗

Immunohistochemical expression of cytokeratins 7 and 20 in malignant salivary gland tumors.

On the basis of the heterogeneity of cytokeratins 7 and 20 expression in malignant epithelial tumors, the cytokeratin 7/20 immunophenotype has served as a useful diagnostic tool for discrimination of primary and/or metastatic carcinomas of unknown origin. However, the expression pattern of these cytokeratins in malignant salivary gland tumors has not been thoroughly studied. Our study material was composed of 84 malignant tumors of primary major or minor salivary gland origin. Nine histologic types of carcinoma were represented, including mucoepidermoid (26 cases), adenoid cystic (25), polymorphous low grade (11), salivary duct (8), acinic cell (4), ex mixed tumor (3), not otherwise specified (3), clear cell (2), and basal cell (2). In all, 13 cases of primary skin or mucosal squamous cell carcinoma with secondary salivary gland involvement were also examined. Immunoreactivity for cytokeratin 7 was evident in all malignant salivary gland tumors; the staining pattern was diffuse and strong in 62 cases, and focal and strong in 22 cases. In contrast, 78 cases were negative for cytokeratin 20, whereas only six cases (two mucoepidermoid, one adenoid cystic, and three salivary duct) displayed focal weak positivity. Overall, 92.9% of malignant salivary gland tumors were characterized by a cytokeratin 7 positive/20 negative immunoprofile, the remaining 7.1% of cases being positive for both cytokeratins. The latter phenotype was more common in salivary duct carcinomas (P< or =0.05). On the other hand, most squamous cell carcinomas (69%) were negative for both cytokeratins, while the remaining cases (31%) were negative for cytokeratin 20 and focally weakly positive for cytokeratin 7. We suggest that assessment of cytokeratin 7/20 immunoprofile may facilitate the differential diagnosis of (a) primary malignant salivary gland tumors from metastatic tumors, (b) metastatic salivary gland tumors, (c) primary salivary gland tumors, especially mucoepidermoid carcinomas, from squamous cell carcinomas, and (d) salivary duct carcinomas from other malignant salivary gland tumors.

Adenocarcinoma↗

IRTA1+ monocytoid B cells in reactive lymphadenitis show a unique topographic distribution and immunophenotype and a peculiar usage and mutational pattern of IgVH genes.

The origin and function of monocytoid B cells (MBCs) are poorly understood. Taking advantage of their strong expression of IRTA1 (a receptor that is also associated with MALT marginal zone B cells), we have comprehensively analysed MBCs in 25 cases of lymphadenitis of different aetiologies, shedding new light on the topographical distribution, immunophenotype and IgV(H) gene usage and mutational profile of this B cell subset. IRTA1(+) MBCs, although predominantly located in the subcapsular and intermediary sinuses, were also observed scattered within germinal centres (GCs) in all lymphadenitis cases examined. The molecular characterization of IgV(H) genes revealed that IRTA1(+) MBCs residing in different areas of the lymph node (subcapsular sinus, intermediary sinuses and GCs) can be clonally related (with intraclonal variation), and that those located in GCs are consistently more mutated and selected for expression of a functional antigen receptor than those located in the sinuses. Moreover, by contrast, IRTA1(+) MBCs in GCs express the memory B cell marker CD27. Finally, in toxoplasmic lymphadenitis, the IRTA1(+) MBC population shows a highly preferential usage of the V(H) genes 3-7 and 3-30 (without any obvious peculiarity in their CDR3s), possibly suggesting that a superantigen expressed by Toxoplasma gondii may be involved in the early activation of this B cell subset.

B-Lymphocyte Subsets↗

Rosacea: current thoughts on origin.

Rosacea is a clinical pattern beginning and evolving in the genetically susceptible individual in response to a host of exposures. It produces a variety of clinical presentations, which vary over time and with age. Recently, many specific mediators of rosacea development have been described. A primary genetic cause for rosacea is suggested as single genes often control such mediators: enzymes, neuroendocrine transmitters, and cytokines are found in pathways to rosacea signs and symptoms. Currently, neither a specific cause nor a laboratory indicator of rosacea has been suggested. However, broadening interest in rosacea portends future increase in knowledge.

Cytokines↗

Characteristics of two cases with dup(15)(q11.2-q12): one of maternal and one of paternal origin.

PURPOSE: The phenotype correlations for interstitial duplications that include the Prader-Willi/Angelman syndrome critical region are not well established. We describe two such duplication cases, one of which was of maternal origin and the other was paternal. METHODS: High resolution G-banding, fluorescence in situ hybridization (FISH) for SNRP-N and D15S10 were used for cytogenetic analysis. Southern blot analyses based on parent of origin specific DNA methylation at D15S63 (PW71) locus were utilized for detection of methylated and unmethylated fragments. RESULTS: The duplication was established by the FISH analysis. The molecular pattern suggested a maternal origin of the duplication in patient 1 and a paternal origin in patient 2. Patient 1 (2 years old) had developmental and speech delays with pervasive developmental disorder or mild autism, strabismus, and normal growth parameters with seizures. Patient 2 (16 years old) had global developmental delay, verbal IQ of 94, depression, obesity, food-seeking behavior, and significant behavioral problems that included self-injurious tendencies. Neither patient had significant dysmorphic features or abnormalities of internal organs. CONCLUSION: The two cases suggest that some patients with 15q11.2q12 duplication may have significant anomalies, and there appear to be phenotypic differences between maternal and paternal transmission of the duplication.

Chromosomes, Human, Pair 15↗

Novel Alterations in Plasmid DNA Associated with Aromatic Hydrocarbon Utilization by Pseudomonas putida R5-3.

Subcultures of Pseudomonas putida R5-3 altered their plasmid DNA content in specific ways depending on the particular aromatic hydrocarbon utilized as the sole carbon source. Two indigenous plasmids, 115 and 95 kilobases (kb) in size, were observed in R5-3A, which was derived from R5-3 by growth on minimal medium containing p-methylbenzoate as the sole carbon source. When R5-3A was transferred to medium containing m-xylene or toluene, derivative strains were obtained in which the 95-kb plasmid was lost and a new plasmid of 50 or 60 kb appeared. Reversion to the original plasmid profile of R5-3A was observed when xylene- or toluene-grown cells were returned to medium containing p-methylbenzoate. Restriction enzyme analysis and Southern blot hybridizations of total plasmid DNA indicated deletions and rearrangements of DNA restriction fragments in the derivatives maintained on m-xylene and toluene when compared with the original R5-3A. In the derivatives which retrieved the original plasmid profile, the restriction enzyme fragment pattern was identical to that in the original R5-3A, in that the fragments which were missing after growth on m-xylene or toluene were again present. Southern blot hybridizations revealed that part of the plasmid DNA lost from the original plasmid profile was integrated into the chromosomal DNA of xylene-grown R5-3B and that these plasmid fragments were associated with aromatic hydrocarbon metabolism. Hybridization with pathway-specific DNA fragments from the TOL plasmid pWWO indicated that this 95-kb plasmid contains DNA homologous to the meta-fission pathway genes.

Journal Article↗

An improved combined cholinesterase stain and silver impregnation method for quantitative analysis of innervation patterns in frog muscle.

The original method combining Karnovsky's cholinesterase stain and Bodian silver impregnation has been modified to stain both myelinated and unmyelinated axons and to reduce background staining. The improvements were obtained by adding nitric acid to a paraformaldehyde-acetone fixative and by carrying out the silver impregnation of axons in an alcoholic solution. The method is especially suitable for quantitative estimation of the different kinds of nerve sprouting as well as for study of the remodeling of neuromuscular junctions in normal and experimental frog muscles.

Animals↗