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61.
De Luca A Buccino A Gianni D Mangino M Giustini S Richetta A Divona L Calvieri S Mingarelli R Dallapiccola B 《Human mutation》2003,21(2):171-172
The high mutation rate at the NF1 locus results in a wide range of molecular abnormalities. The majority of these mutations are private and rare, generating elevated allelic diversity with a restricted number of recurrent mutations. In this study, we have assessed the efficacy of denaturing high-performance liquid chromatography (DHPLC), for detecting mutation in the NF1 gene. DHPLC is a fast and highly sensitive technique based on the detection of heteroduplexes in PCR products by ion pair reverse-phase HPLC under partially denaturing conditions. We established theoretical conditions for DHPLC analysis of all coding exons and splice junctions of the NF1 gene using the WAVEmaker software version 4.1.40 and screened for mutations a panel of 40 unrelated NF1 patients (25 sporadic and 15 familial), genetically uncharacterized. Disruptive mutations were identified in 29 individuals with an overall mutation detection rate of 72.5%. The mutations included eight deletions (exons 4b, 7, 10a, 14, 26, and 31), one insertion (exon 8), nine nonsense mutation (exons 10a, 13, 23.1, 27a, 29, 31, and 36), six missense mutations (exons 15, 16, 17, 24, and 31), four splice errors (exons 11, 14, 36, and 40) and a complex rearrangement within exon 16. Eighteen (62%) of the identified disruptive mutations are novel. Seven unclassified and three previously reported polymorphisms were also detected. None of the missense mutations identified in this study were found after screening of 150 controls. Our results suggest that DHPLC provides an accurate method for the rapid identification of NF1 mutations. 相似文献
62.
Peter Gross Will W. Minuth Markus Ketteler Eberhard Frömter Rita Böhm 《Pflügers Archiv : European journal of physiology》1988,412(4):434-441
The ionic conductive properties were studied of epithelia of collecting duct principal cells which had been grown in primary tissue culture from renal cortex/capsule explants. When pretreated with aldosterone (10–6 mol/l) and bathed on either surface with isotonic HCO
3
–
-free Ringer's solution, the transepithelial voltage,V
te, varied between –21 and –72 mV (apical surface negative) while the transepithelial resistance,R
te, ranged from 0.4 to 1.5 kcm2. By 10:1 step-changes in Na+ concentration the apical cell membrane was shown to have a high conductivity for sodium, inhibitable by amiloride, 10–6 mol/l. However, contrary to observations in natural collecting duct under control conditions, amiloride never reversed the polarity ofV
te even at 10–4 mol/l. Both the apical and the basolateral cell membranes were conductive for potassium and both conductivities were inhibitable by Ba2+ (5 mmol/l). 10:1 reduction of apical Cl– concentration strongly hyperpolarizedV
te with a monophasic time course suggesting the presence of a paracellular shunt conductance for Cl–. In addition there may be a small Cl– conductance present in the apical cell membrane since apical application of the chloride channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPAB) at 10–7 mol/l produced a minute but significant hyperpolarization. On the other hand, 10:1 reduction of basolateral Cl– concentration caused a biphasic change inV
te (initial depolarization, followed by repolarization) which indicates the presence of a large Cl– conductance in the basolateral cell membrane. The latter was not inhibitable by 10–7 mol/l NPPAB. Higher concentrations of this and of an other Cl– channel blocker produced non-specific effects. In conclusion, our studies of a pure principal cell epithelium confirm findings described for the intact cortical collecting duct and add new information concerning chloride conductivity and related blocking agents.Dedicated to Prof. Dr. H. Sitte, Homburg, FRG, upon his 60th birthday. 相似文献
63.
Giant cell tumor of bone express p63. 总被引:1,自引:0,他引:1
Brendan C Dickson Shu-Qiu Li Jay S Wunder Peter C Ferguson Behnam Eslami Joel A Werier Robert E Turcotte Rita A Kandel 《Modern pathology》2008,21(4):369-375
p63 contributes to skeletal development and tumor formation; however, little is known regarding its activity in the context of bone and soft tissue neoplasms. The purpose of this study was to investigate p63 expression in giant cell tumor of bone and to determine whether it can be used to discriminate between other giant cell-rich tumors. Seventeen cases of giant cell tumor of bone were examined to determine the cell type expressing p63 and identify the isoforms present. Total RNA or cell protein was extracted from mononuclear- or giant cell-enriched fractions or intact giant cell tumor of bone and examined by RT-PCR or western blot, respectively. Immunohistochemistry was used to evaluate p63 expression in paraffin embedded sections of giant cell tumor of bone and in tumors containing multinucleated giant cells, including: giant cell tumor of tendon sheath, pigmented villonodular synovitis, aneurysmal bone cyst, chondroblastoma, and central giant cell granuloma. The mononuclear cell component in all cases of giant cell tumor of bone was found to express all forms of TAp63 (alpha, beta, and gamma), whereas only low levels of the TAp63 alpha and beta isoforms were detected in multinucleated cells; DeltaNp63 was not detected in these tumors. Western blot analysis identified p63 protein as being predominately localized to mononuclear cells compared to giant cells. This was confirmed by immunohistochemical staining of paraffin-embedded tumor sections, with expression identified in all cases of giant cell tumor of bone. Only a proportion of cases of aneurysmal bone cyst and chondroblastoma showed p63 immunoreactivity whereas it was not detected in central giant cell granuloma, giant cell tumor of tendon sheath, or pigmented villonodular synovitis. The differential expression of p63 in giant cell tumor of bone and central giant cell granuloma suggest that these two tumors may have a different pathogenesis. Moreover, p63 may be a useful biomarker to differentiate giant cell tumor of bone from central giant cell granuloma and other giant cell-rich tumors, such as giant cell tumor of tendon sheath and pigmented villonodular synovitis. 相似文献
64.
Ghiorzo P Villaggio B Sementa AR Hansson J Platz A Nicoló G Spina B Canepa M Palmer JM Hayward NK Bianchi-Scarrà G 《Human pathology》2004,35(1):25-33
Little is known about the correlation between the loss of p16 expression and tumor progression in familial melanoma; no systematic study has been conducted on p16 expression in melanocytic tumors from patients carrying germline CDKN2A mutations. We analyzed 98 early primary lesions from familial patients, previously tested for germline CDKN2A status, by quantitative immunohistochemistry using 3 p16 antibodies. We found that p16 expression was inversely correlated with tumor progression and was significantly lower in melanomas, including in situ lesions, than in nevi. Of other features analyzed, tumor thickness showed the most significant correlation with p16 levels. Lesions from mutation-negative patients displayed combined nuclear and cytoplasmic staining. However, some mutation-positive lesions (ie, G101W, 113insR, M53I, R24P, and 33ins24), including benign nevi, showed nuclear mislocalization, confirming previous studies suggesting that subcellular distribution indicates functional impairment of p16. 相似文献
65.
Chalupníková R 《Acta medica (Hradec Králové). Supplementum Universitas Carolina, Facultas Medica Hradec Králové》2004,47(2):127-130
This article is aimed at developing of multimedia teaching programs. Authoring tool Authorware Professional for Windows is briefly described as a suited system for the developing of didactic applications. Author presents own application based on Authorware--Biophysics of perception of sound and light. At the end are also disscused benefits of multimedia teaching tools. 相似文献
66.
Phenotypic and genotypic analyses of enterohemorrhagic Escherichia coli O145 strains from patients in Germany 总被引:5,自引:0,他引:5 下载免费PDF全文
Sonntag AK Prager R Bielaszewska M Zhang W Fruth A Tschäpe H Karch H 《Journal of clinical microbiology》2004,42(3):954-962
Enterohemorrhagic Escherichia coli (EHEC) strains of serogroup O145 are emerging as causes of diarrhea and the hemolytic-uremic syndrome. However, there have been few genetic analyses of this EHEC group. We investigated the serotypes, virulence genes, plasmid profiles, pulsed-field gel electrophoresis (PFGE) patterns, and genetic variability of the fliC and eae genes in 120 EHEC O145 strains isolated from cases of hemolytic-uremic syndrome (n = 24) or diarrhea (n = 96) in Germany between 1996 and 2002. Three isolates belonged to serotype O145:H28, one to serotype O145:H25, and 116 were nonmotile (O145:H(-)). One hundred fourteen of the nonmotile strains shared fliC restriction fragment length polymorphism (RFLP) patterns identical to that of the O145:H28 strains. The remaining two nonmotile strains displayed a fliC-RFLP pattern identical to that of the O145:H25 strain. Each of the 117 strains with the fliC-RFLP(H28) pattern harbored eae gamma, whereas the three strains with the fliC-RFLP(H25) pattern possessed eae beta. Five different stx genotypes, six combinations of plasmid-encoded putative virulence genes, 29 plasmid profiles, and 47 PFGE types were identified. Strains within some of the PFGE types could be further subtyped by means of distinct plasmid profiles. These data demonstrate that the EHEC O145 serogroup is comprised of two different serotypes that possess distinct eae types. The heterogeneity of EHEC O145 strains at the chromosomal and plasmid level, in particular the high diversity in PFGE patterns, provides a basis for molecular subtyping of these pathogens. 相似文献
67.
68.
Francesco Muntoni Anna Mateddu Maria Giovanna Marrosu Miiena Cau Rita Congiu Maria Antonietta Melis Antonio Cao Carlo Cianchetti 《Clinical genetics》1992,42(1):35-38
The majority of Duchenne muscular dystrophy (DMD) female carriers show dystrophin immunostaining abnormalities, although a significant proportion of clinically non-manifesting carriers are normal following this analysis. We had the opportunity to study dystrophin immunostaining in two different muscles, the vastus lateralis and the rectus abdominis of a possible DMD carrier. While the vastus showed normal dystrophin immunostaining, pathological staining was detected in her rectus abdominis. These findings seem to indicate that dystrophin expression can vary in different muscle groups of a DMD carrier. The implications of these findings in DMD carrier detection and possible dystrophin function are discussed. 相似文献
69.
70.
Hussey SM Gander R Southern P Hoang MP 《Archives of pathology & laboratory medicine》2005,129(6):794-797
Primary subcutaneous phaeohyphomycosis can rarely be caused by Cladophialophora bantiana, and we present the histologic and culture findings of such a case. A 32-year-old African American woman with systemic lupus erythematosus presented with a 2-year history of multiple, recurrent, tender, and ulcerated skin nodules with purulent drainage on her upper back. Histologic sections of the excision demonstrated features of phaeohyphomycosis. Culture findings were characteristic of C bantiana. Of interest, at age 10 she had sustained traumatic implantation of wood splinters into this area during a tornado, yet clinical symptoms of a subcutaneous infection did not manifest until she developed lupus erythematosus at age 27. Our case highlights the role of trauma and immunosuppression in the pathogenesis of subcutaneous phaeohyphomycosis. 相似文献