Effect of ampicillin-sulbactam on clinical capillary zone electrophoresis of serum proteins

Background: Capillary zone electrophoresis (CZE) is a well-accepted automated method used to separate serum proteins and detect monoclonal components. CZE uses ultraviolet detection at 214nm to directly quantify proteins via peptide bonds. Any substance that absorbs at 214nm and is present in serum can potentially interfere with CZE analysis. This has been reported for radio-contrast media and antibiotics. Methods: Here we describe a peak on the anode side of the α2-globulin fraction caused by the antibiotic ampicillin-sulbactam (Unacid®). Results and conclusions: Extra peaks that can be misinterpreted as monoclonal components can be present in almost all electrophoretic fractions of CZE. Immunosubtraction or immunofixation is always required to rule out these conditions. Clin Chem Lab Med 2008;46:1468-

Die quantitative Analyse von Markerproteinen im Urin Quantitative analysis of marker proteins in urine

Die Analyse von spezifischen Proteinen im zweiten Morgenurin, bezogen auf den Kreatiningehalt der Probe, erlaubt heute nicht nur den Nachweis oder den Ausschluss von Nierenerkrankungen, sondern darüber hinaus auch die Differenzierung und Verlaufskontrolle von Nephropathien. Störungen lassen sich aufgrund ihres Markerproteinprofils in solche mit hauptsächlich glomerulärem oder tubulärem Anteil und zusätzlich in weitere Untergruppen einteilen. Im Zusammenhang mit den Teststreifenresultaten kann die Quelle einer Blutung mit spezifischen Quotienten näher eingegrenzt und Kontaminationen können von tatsächlichen renalen Proteinurien unterschieden werden. Eine Plausibilitätsprüfung und Interpretation der erhaltenen Ergebnisse ist unbedingt erforderlich. Da eineVielzahl von Regeln überprüft werden muss, ist eine Berechnung und Darstellung der Ergebnisse nur mit Hilfe von wissensbasierten Systemen in Kombination mit einer grafischen Befunddarstellung sinnvol

Effect of ampicillin-sulbactam on clinical capillary zone electrophoresis of serum proteins.

BACKGROUND: Capillary zone electrophoresis (CZE) is a well-accepted automated method used to separate serum proteins and detect monoclonal components. CZE uses ultraviolet detection at 214 nm to directly quantify proteins via peptide bonds. Any substance that absorbs at 214 nm and is present in serum can potentially interfere with CZE analysis. This has been reported for radio-contrast media and antibiotics. METHODS: Here we describe a peak on the anode side of the alpha(2)-globulin fraction caused by the antibiotic ampicillin-sulbactam (Unacid). RESULTS AND CONCLUSIONS: Extra peaks that can be misinterpreted as monoclonal components can be present in almost all electrophoretic fractions of CZE. Immunosubtraction or immunofixation is always required to rule out these conditions

Evaluation of proteinuria and GFR to diagnose and classify kidney disease : systematic review and proof of concept

Chronic kidney disease is often not associated with significant symptoms or abnormalities in common laboratory test results. Diagnosis is supposedly facilitated by calculating the glomerular filtration rate (GFR) from serum creatinine. A reference range GFR, however, does not exclude renal disease, because renal disease causes the subsequent decrease of renal function. Thorough analysis of proteinuria, however, requires a profound knowledge of the renal handling of the different marker proteins of glomerular and tubular origin. This paper summarizes the scientific basis, explains the diagnostic rationale and proves the concept by analyzing 5669 samples, where GFR and proteinuria work-up were available. 63% (1446 of 2287) of the samples with a GFR above 60 showed either glomerular (37.8%, n=865) or tubular proteinuria (25.4%, n= 581). The quantity of proteinuria increased severely with decreasing kidney function. The rate of glomerular proteinuria remained nearly constant in the different GFR groups, while primarily tubular proteinuria increased from 23% to 63%. A proteinuria pattern indicating a good response to therapy was frequently combined with a high GFR (selective glomerular proteinuria/ incomplete tubular proteinuria), while the severe forms of unselective or complete tubular proteinuria associated with a severe GFR decrease. Regression analysis showed a better inverse correlation of GFR with tubular (r=-0.643) than glomerular markers (r=-0.360; combined r=-0.646). We believe that this complex interrelated laboratory information must be delivered most effectively, i.e. with the use of a knowledge based system in combination with improved, visual oriented laboratory output

면역화학측정법을 이용한 혈청 Transferrin 측정에 관한 연구

Various methods are available for measuring total iron·binding capacity in serum (TIBC) which is an indirect measure of serum trans· ferrin content. All of these methods are cumbersome and have methodological disadvantages such as a high sample amount , possibility for iron contamination of the laboratory ware unspecificity because of other iron-binding proteins in serum and problems to optimize the method (Koepke, 1965; Williams and Conrad, 1972; Von der Heul et aI., 1972; Frazer, 197:1; Haeckel et aI. , 1973; Schmidt et aI. , 1975; Tsung ct al 1975; Graham and Bates, 1976; Rajamacki ct aI., 1979; Seiffert, 1981). Immunochemical methods have made a direct immunological measurement of serum transferrin possible (Goodman et aI., 1958; Wilding and Rollason, 1972; Haeckel et aI., 1973; Schmidt et aI., 1975; Tsung et aI., 1975; Kreutzer. 1976) . The purpose of the present work was to study the correlation hetween serum transferrin and TIBC values in healthy and anemic persons. * This research was supported in part by scholarship from the State Government for Bessen in Federal R('public of Germany in the year of 10m Further to determine reference values of serum transferrin for adults of both sexes and for 2 groups of children at different age

A modern approach to CSF analysis: pathophysiology, clinical application, proof of concept and laboratory reporting.

The CNS immune response often leads to characteristic interrelated biochemical changes in cerebrospinal fluid. Multiple analytes, i.e. cell count, cell differential, evaluation of barrier function and intrathecal IgG, IgA and IgM synthesis should be included in basic diagnostic workup. We describe the scientific background, laboratory investigations and characteristic patterns found with basic CSF analysis, based on the recommendations of the German cerebrospinal fluid society. The concept is substantiated by retrospectively analyzing data of 4026 paired CSF/serum samples. 53% of our samples presented with at least one or several combined abnormal findings. An intrathecal IgG, IgA or IgM immunoglobulin response (37%, n=1481) and a blood-CSF barrier dysfunction (37%; n=1473) were most frequent; followed by an elevated leukocyte cell count (25%; n=992). The immunoglobulin response showed an intrathecal production of IgG in 49% (n=731/1481), which was only detectable in isoelectric focusing in 27% (n=200/731). Intrathecal IgM (n=389) and IgA (n=361) synthesis presented with nearly equal frequency of 25% in samples with intrathecal immunoglobulin response. The immunoglobulin pattern showed a solitary one class reaction of IgG, IgA or IgM in 67%, a combined two class reaction of IgG/IgA, IgG/IgM or IgA/IgM synthesis in 16% and a combined three-class reaction of IgG, IgA and IgM in 17%. This approach generates valuable but numerous complex and interrelated biochemical data. We therefore developed a knowledge-based system combined with visual oriented laboratory output to transfer the information more effectively. This often uncovers typical patterns specific for distinct neurological diseases, is well accepted by our medical community documented by a 37% increase in external ordering

HDF1 and RAD17 Genes are Involved in DNA Double-strand Break Repair in Stationary Phase Saccharomyces cerevisiae

DNA repair, checkpoint pathways and protection mechanisms against different types of perturbations are critical factors for the prevention of genomic instability. The aim of the present work was to analyze the roles of RAD17 and HDF1 gene products during the late stationary phase, in haploid and diploid yeast cells upon gamma irradiation. The checkpoint protein, Rad17, is a component of a PCNA-like complex—the Rad17/Mec3/Ddc1 clamp—acting as a damage sensor; this protein is also involved in double-strand break (DBS) repair in cycling cells. The HDF1 gene product is a key component of the non-homologous end-joining pathway (NHEJ). Diploid and haploid rad17Δ/rad17Δ, and hdf1Δ Saccharomyces cerevisiae mutant strains and corresponding isogenic wild types were used in the present study. Yeast cells were grown in standard liquid nutrient medium, and maintained at 30°C for 21 days in the stationary phase, without added nutrients. Cell samples were irradiated with 60Co γ rays at 5 Gy/s, 50 Gy ≤ Dabs ≤ 200 Gy. Thereafter, cells were incubated in PBS (liquid holding: LH, 0 ≤ t ≤ 24 h). DNA chromosomal analysis (by pulsed-field electrophoresis), and surviving fractions were determined as a function of absorbed doses, either immediately after irradiation or after LH. Our results demonstrated that the proteins Rad17, as well as Hdf1, play essential roles in DBS repair and survival after gamma irradiation in the late stationary phase and upon nutrient stress (LH after irradiation). In haploid cells, the main pathway is NHEJ. In the diploid state, the induction of LH recovery requires the function of Rad17. Results are compatible with the action of a network of DBS repair pathways expressed upon different ploidies, and different magnitudes of DNA damage

Role of UME6 in transcriptional regulation of a DNA repair gene in Saccharomyces cerevisiae

In Saccharomyces cerevisiae UV radiation and a variety of chemical DNA-damaging agents induce the transcription of specific genes, including several involved in DNA repair. One of the best characterized of these genes is PHR1, which encodes the apoenzyme for DNA photolyase. Basal-level and damage-induced expression of PHR1 require an upstream activation sequence, UAS(PHR1), which has homology with DRC elements found upstream of at least 19 other DNA repair and DNA metabolism genes in yeast. Here we report the identification of the UME6 gene of S. cerevisiae as a regulator of UAS(PHR1) activity. Multiple copies of UME6 stimulate expression from UAS(PHR1) and the intact PHR1 gene. Surprisingly, the effect of deletion of UME6 is growth phase dependent. In wild-type cells PHR1 is induced in late exponential phase, concomitant with the initiation of glycogen accumulation that precedes the diauxic shift. Deletion of UME6 abolishes this induction, decreases the steady-state concentration of photolyase molecules and PHR1 mRNA, and increases the UV sensitivity of a rad2 mutant. Despite the fact that UAS(PHR1) does not contain the URS1 sequence, which has been previously implicated in UME6-mediated transcriptional regulation, we find that Ume6p binds to UAS(PHR1) with an affinity and a specificity similar to those seen for a URS1 site. Similar binding is also seen for DRC elements from RAD2, RAD7, and RAD53, suggesting that UME6 contributes to the regulated expression of a subset of damage-responsive genes in yeast