Zusammenfassung Individuelle normale IgG wurden isoelektrisch in eine Vielzahl von Komponenten getrennt, deren IP zwischen pH 5,5 und 9,5 lagen. Im Gegensatz dazu focussierten individuelle Myelom IgG einheitlich. Die isoelektrischen Punkte dieser Paraproteine unterschieden sich untereinander stark, verteilten sich alle zusammen jedoch ebenfalls über den pH-Bereich 5–9. Dieser Befund entspricht der Auffassung, daß diese Paraproteine normale Immunglobuline repräsentieren.Ähnliche Ergebnisse hatte die Fraktionierung der L- und H-Polypeptidketten. Während normale L- und H-Ketten eine ausgeprägte Heterogenität aufwiesen, waren die IgG-L- und H-Ketten von Myelomproteinen einheitlich. Mittels gleichzeitiger Anwendung der PAA-Elektrophorese, quantitativen Aminosäurenanalyse, Endgruppenbestimmung und anderer protein-chemischer Methoden konnte gezeigt werden, daß im Falle der L-Ketten sowohl die Aminosäureverteilung als auch die Konfiguration das unterschiedliche isoelektrische Verhalten bedingt. Die Anwendung der isoelektrischen Analyse für den Nachweis von Rekombinatbildungen aus L- und H-Ketten wurde gezeigt.
Summary Myeloma IgG and its light and heavy polypeptide chains were subjected to isoelectric focusing in a vertical column containing a combined pH and density gradient. The results have been compared to those obtained by isoelectric separation of normal individual IgG and its polypeptide chains. In carrier ampholytes of two pH steps each individual myeloma globulin was focused in only one fraction, but the isoelectric points of all myeloma globulins were distributed between pH 5 and pH 9. In contrast to these results each normal individual IgG was electrofocused in a multiplicity of isoelectric fractions exhibiting the same pH range from pH 5 to pH 9. Similar results were obtained by the analysis of light and heavy polypeptide chains. A remarkable degree of heterogeneity could be demonstrated in normal and a homogenous isoelectric pattern in myeloma IgG polypepide chains. By polyacrylamide-gel-electrophoresis it could be demonstrated that a direct proportionality of isoelectric point and electrophoretic mobility in polyacrylamide-gel did'nt exist. By the quantitative aminoacid determination, endgroup analysis and other physico-chemical methods it could be shown, that differences in the isoelectric point of L-chains are not exclusively caused by differences in the amino acid composition. The use of isoelectric focusing for the analysis of L- and H-chains recombination has been demonstrated.
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Osteonecrosis of the femoral head was produced in rats by cutting the ligamentum teres and incising the cervical periosteum. As of the second postoperative week, fibrous tissue pervaded the necrotic epiphyses, macrophages and osteoclasts removed the debris, osteoblasts deposited lamellar-fibred and woven-fibred intramembranous bone, and remodeling began. In 16% of the rats killed during the 2nd postoperative week, the epiphyses contained big fragments of necrotic bone enclosed by densely packed, capillary-sized vessels. Ingrowth of this hypervascularized, pyogenic granuloma-like tissue is presumably due to the presence of excessive growth factors, reflecting an exaggerated pathophysiological reaction within the framework of organization of the necrotic epiphyses. 相似文献
Composite bone cements were formulated with bioactive glass (MgO--SiO(2)--3CaO. P(2)O(5)) as the filler and hydrophilic matrix. The matrix was composed of a starch/cellulose acetate blend (SCA) as the solid component and a mixture of methylmethacrylate/acrylic acid (MMA/AA) as the liquid component. The curing parameters, mechanical properties, and bioactive behavior of these composite cements were determined. The addition of up to 30 wt % of glass improved both compressive modulus and yield strength and kept the maximum curing temperature at the same value presented by a typical acrylic-based commercial formulation. The lack of a strongly bonded interface (because no coupling agent was used) had important effects on the swelling and mechanical properties of the novel bone cements. However, bone cements containing AA did not show a bioactive behavior, because of the deleterious effect of this monomer on the calcium phosphate precipitation on the polymeric surfaces. Formulations without AA were prepared with MMA or 2-hydroxyethyl methacrylate (HEMA) as the liquid component. Only these formulations could form an apatite-like layer on their surface. These systems, therefore, are very promising: They are bioactive, hydrophilic, partially degradable, and present interesting mechanical properties. This combination of properties could facilitate the release of bioactive agents from the cement, allow bone ingrowth in the cement, and induce a press-fitting effect, improving the interfaces with both the prosthesis and the bone. 相似文献
Glioblastomas develop de novo (primary glioblastomas) or through progression from low-grade or anaplastic astrocytoma (secondary glioblastomas). There is increasing evidence that these glioblastoma subtypes develop through different genetic pathways. Primary glioblastomas are characterized by EGFR and MDM2 amplification/overexpression, PTEN mutations, and p16 deletions, whereas secondary glioblastomas frequently contain p53 mutations. Loss of heterozygosity (LOH) on chromosome 10 (LOH#10) is the most frequent genetic alteration in glioblastomas; the involvement of tumor suppressor genes, other than PTEN, has been suggested. We carried out deletion mappings on chromosome 10, using PCR-based microsatellite analysis. LOH#10 was detected at similar frequencies in primary (8/17; 47%) and secondary glioblastomas (7/13; 54%). The majority (88%) of primary glioblastomas with LOH#10 showed LOH at all informative markers, suggesting loss of the entire chromosome 10. In contrast, secondary glioblastomas with LOH#10 showed partial or complete loss of chromosome 10q but no loss of 10p. These results are in accordance with the view that LOH on 10q is a major factor in the evolution of glioblastoma multiform as the common phenotypic end point of both genetic pathways, whereas LOH on 10p is largely restricted to the primary (de novo) glioblastoma. 相似文献
We report here a polymerase chain reaction (PCR)-based DNA profiling technique that permits Trypanosoma cruzi strain characterization by direct study of infected tissues. This is based on application of a recently developed method of DNA fragment identification, called low-stringency single specific primer PCR (LSSP-PCR), to the study of the variable region of kinetoplast DNA (kDNA) minicircles from T. cruzi Thus, we can translate the intraspecific polymorphism in the nucleotide sequence of kDNA minicircles into a specific and highly reproducible kDNA signature. Comparison with the phenogram obtained by DNA fingerprinting analysis of a set of T. cruzi strains showed good qualitative correlation between the degree of divergence of the LSSP-PCR profiles and the genetic distance between the strains. kDNA signatures of heart tissue from acutely or chronically infected animals revealed perfect concordance with the patterns obtained from cultured parasites for the CL and Colombiana strains but not for the Y strain, which is known to be multiclonal. However, the match was perfect for studies with two clones of the Y strain. We take this as evidence that in some multiclonal strains there is heterogeneity among the clones in the degree of tropism for the heart tissue. Finally, we showed that it is possible to obtain a T. cruzi kDNA signature from the heart of a human patient with chronic Chagasic myocardiopathy. kDNA signatures obtained by LSSP-PCR of sequences amplified from infected tissues constitute a new tool to study the molecular epidemiology of Chagas' disease. 相似文献
Head and neck squamous cell carcinoma is a disease associated with tobacco and alcohol abuse. There is evidence that the oncogenic human papillomavirus (HPV) may also be a risk for upper aerodigestive tract cancers. High-risk HPVs encode two early proteins, E6 and E7, that can bind to p53 and pRb, respectively, and induce its degradation or inactivation. The TP53 gene has a single polymorphism at codon 72 of exon 4 that encodes either arginine (Arg) or proline (Pro). The purpose of this study was to evaluate the role of HPV infection and TP53 polymorphism in head and neck cancer. We analyzed 50 tumors, as well swabs of oral mucosa from 142 control individuals, with a polymerase chain reaction technique. The prevalence of HPV in controls was 10.6% and in cancer specimens 16%. The frequency distribution of genotypes in controls was 50% Arg/Arg, 43% Arg/Pro and 7% Pro/Pro; in tumors, it was 52% Arg/Arg, 32% Arg/Pro, and 16% Pro/Pro. Contrary to the results of some studies on cervical cancer, no association between any TP53 genotype or allele and the development of head and neck cancer was observed, regardless of HPV status, except for the Pro/Pro genotype, which is associated with the absence of HPV. The arginine allele appears to protect against head and neck cancers. Also, the data showed that HPV infection results in no increased risk of developing head and neck tumors. 相似文献
OBJECTIVES: To use two rapid human immunodeficiency virus (HIV) tests at labor, measure test acceptance and performance, and measure HIV prevalence in these women. METHODS: Between February and October 2000, two rapid tests (Determine; Abbott, Chicago, IL, U.S.A. and Double Check; Orgenics, Yavne, Israel) were used in three public maternities in Rio de Janeiro, Brazil. Enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) analysis confirmed positive and discordant results. RESULTS: Of the 858 patients who were enrolled, the mean gestational age was 36 weeks (median = 39, mode = 40) and 17 (2%) refused testing. Of the 841 patients tested, 13 were positive by both tests, which represents a 1.5% prevalence (95% confidence interval: 0.7%-2.3%); all were confirmed by ELISA and WB analysis. Seven samples gave discordant results by the rapid tests; of these, six were ELISA-negative/WB-negative and one was ELISA-negative/WB-indeterminate. The positive predictive value for samples that were positive by both rapid tests simultaneously was 100%. CONCLUSIONS: Two rapid HIV tests used at labor were well accepted (98%). When the combined results of the two rapid tests (but not a single rapid test) were analyzed, this strategy was as efficient as the standard ELISA and WB HIV strategy for correctly classifying individuals. 相似文献
Introduction: Neuropsychiatric systemic lupus erythematosus (NPSLE) is characterized by a heterogeneity of clinical manifestations. The absence of diagnostic criteria and the lack of clinical trials is a challenge in clinical practice.
Areas covered: A literature review was performed to describe epidemiology, characterization (clinical, immunological, and imaging), diagnosis and treatment of NPSLE. Classification criteria have been the first step towards a uniform definition. More recently, different attribution models have been developed to help to determine if the NP event is due to SLE. Disease activity is a major risk factor for NP events. Cytokines and autoantibodies are associated with NP events, however, only a few studies have identified risk factors for individual NP events.
Expert opinion: Further research needs to search for and validate biomarkers for NPSLE and individual NP events, including neuroimaging findings, attribution models, and serologic markers. This will be a fundamental step in planning randomized control trials in the treatment of NPSLE to improve outcome. 相似文献