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Selected AbstractsAlbumin enhanced morphometric image analysis in CLL,CYTOMETRY, Issue 1 2004Matthew A. Lunning Abstract BACKGROUND The heterogeneity of lymphocytes from patients with chronic lymphocytic leukemia (CLL) and blood film artifacts make morphologic subclassification of this disease difficult. METHODS We reviewed paired blood films prepared from ethylene-diamine-tetraacetic acid (ETDA) samples with and without bovine serum albumin (BSA) from 82 CLL patients. Group 1 adhered to NCCLS specifications for the preparations of EDTA blood films. Group 2 consisted of blood films containing EDTA and a 1:12 dilution of 22% BSA. Eight patients were selected for digital photomicroscopy and statistical analysis. Approximately 100 lymphocytes from each slide were digitally captured. RESULTS The mean cell area ± standard error was 127.8 ,m2 ± 1.42 for (n = 793) for group 1 versus 100.7 ,m2 ± 1.39 (n = 831) for group 2. The nuclear area was 88.9 ,m2 ± 0.85 for group 1 versus 76.4 ,m2 ± 0.83 for group 2. For the nuclear transmittance, the values were 97.6 ± 0.85 for group 1 and 104.1 ± 0.83 for group 2. The nuclear:cytoplasmic ratios were 0.71 ± 0.003 for group 1 and 0.78 ± 0.003 for group 2. All differences were statistically significant (P < 0.001). CONCLUSIONS BSA addition results in the reduction of atypical lymphocytes and a decrease in smudge cells. BSA also decreases the lymphocyte area and nuclear area, whereas nuclear transmittance and nuclear:cytoplasmic ratio are increased. A standardized method of slide preparation would allow accurate interlaboratory comparison. The use of BSA may permit better implementation of the blood film-based subclassification of CLL and lead to a better correlation of morphology with cytogenetics and immunophenotyping. Published 2003 Wiley-Liss, Inc. [source] Assessment of Carotid Compliance Using Real Time Vascular Ultrasound Image Analysis in Marfan SyndromeECHOCARDIOGRAPHY, Issue 4 2009Anatoli Kiotsekoglou M.D. Background: Fibrillin-1 deficiency, dysregulated cytokine transforming growth factor-,, and increased collagen deposition related to fibrillin-1 gene mutations could predispose to impaired carotid compliance (CC) in Marfan syndrome (MFS). We sought to detect any alterations in CC using the vascular image analysis system (VIA). Methods and Results: Thirty-two MFS patients, 20 men and 12 women (mean age 34.2 ± 12.05 years), and 29 controls matched for age, sex, and body surface area (BSA) were recruited. The entire length of each carotid system was initially scanned longitudinally using a 14 MHz linear transducer. Then, a stereotactic clamp held the transducer in contact with the carotid artery. Arterial diameter changes during the cardiac cycle were recorded for 1 minute from both right (RCCA) and left common carotid arteries (LCCA) separately using the VIA system. RCCA and LCCA compliance and distensibility measurements were significantly reduced in MFS patients when compared to controls, P < 0.05. RCCA and LCCA intima-media thickness did not differ between patients and controls, P > 0.05. MFS diagnosis and age were associated with reduced CC in both carotid arteries after adjusting for variables such as, sex, BSA, heart rate, beta-blockade, intima-media thickness, and aortic root size. Conclusions: Our findings showed a reduction in CC in adult patients with MFS. This could be attributed to fibrillin-1 deficiency resulting in structural abnormalities in the carotid arterial wall. [source] Echocardiographic Left Ventricular Mass in a Multiethnic Southeast Asian Population: Proposed New Gender and Age-Specific NormsECHOCARDIOGRAPHY, Issue 8 2008M.R.C.P., Raymond Ching-Chiew Wong M.B.B.S. Background: Left ventricular mass (LVM) is an independent risk factor for cardiovascular outcome. We aimed to define normal reference values of LVM/body surface area (BSA) in a multiethnic Southeast Asian population across ages, and define demographic parameters that predict LVM/BSA. Methods: 198 subjects (44% men, mean age 40 ± 14 years, 82% Chinese, 13% Malay and 5% Indian) with no cardiovascular comorbidity and had normal echo images for age were included in the analysis. Echo LVM was calculated as: 1.04 ×[(left ventricular internal diameter at end-diastole {LVIDd}+ interventricular septal thickness at end-diastole {IVSd}+ left ventricular posterior wall thickness at end-diastole {LVPWd})3, LVIDd3× 0.8]+ 0.61, indexed by BSA (LVM/BSA)* and expressed as g/m2. Results: BSA and blood pressure (BP) were comparable between dichotomous age groups < or , 50 years within the same gender. Women aged , 50 years had larger IVSD, LVPWd, LVM and LVM/BSA compared to younger cohort. (p < 0.01 for all variables). The 95th percentile of LVM in men and women were 189 g and 148 g respectively; corresponding values for LVM/BSA were 106 and 96 g/m2. These values are consistently smaller than published values from the West. Age (r = 0.27, P < 0.001), gender (r =,0.30, P < 0.001), and systolic BP (r = 0.25, P = 0.003) were significant univariate predictors of LVM/BSA. Conclusion: We therefore propose a different cutoff value for the diagnosis of LV hypertrophy among Southeast Asians. [source] Reference Values Describing the Normal Mitral Valve and the Position of the Papillary MusclesECHOCARDIOGRAPHY, Issue 7 2007Petrus Nordblom M.Sc. In patients with functional mitral regurgitation (MR), the principal mechanisms are insufficient coaptation due to dilatation of the mitral annulus (MA), global ventricular dysfunction with tethering of leaflets, or restricted leaflet motion with incorrect apposition due to regional ventricular dysfunction and displacement of the papillary muscles (PMs). These different entities often coexist and for this reason, knowledge of the normal reference values describing the shape and size of the MA and the position of the PMs is essential. In the present study, we describe the MA dimensions and the position of the PMs in a group of normal individuals (n = 38, 60% women, age [mean ± SD] 51 ± 9 years and BSA 1.83 ± 0.16 m2) investigated with transthoracic echocardiography. The anteroposterior dimension (AP) of the ellipse-shaped MA was measured in a parasternal long axis, while the distance from the posteromedial (PoM) to the anterolateral (AL) commissure was measured in a parasternal short axis (CC). The annular area was calculated assuming elliptic geometry. The MA shape was described by the ratios AP/CC and AP/length of the anterior leaflet. The PMs' position was described by the following distances: (a) from the MA to the tip of the PoM and AL, PMs measured in a modified two-chamber view where both PMs could be identified, (b) the interpapillary distance, and (c) the tethering distance from the tip of the PM to the contralateral MA. These data on the normal mitral valve morphology should provide useful information when assessing the underlying mechanism of functional MR. [source] Voltammetric Studies of the Interactions Between Ferrocene-Labeled Glutathione and Proteins in Solution or Immobilized onto SurfaceELECTROANALYSIS, Issue 16 2009Yong Peng Abstract Glutathione (GSH) tagged with a ferrocene (Fc) label at its C-terminal was synthesized via coupling ferrocenyl amine to glutathione using o -(benzotriazol-1-yl)- N,N,N,,N, -tetramethyluronium (HBTU)/1-hydroxybenzotrizole (HOBt). The presence of Fc yielded well defined voltammetric signals, rendering the Fc-tagged GSH (GSH-Fc) suitable for electrochemical studies of GSH binding to other biological species. The interaction of GSH-Fc with bovine serum albumin (BSA) was investigated, and a binding ratio of 1.41±0.06 (GSH-Fc/BSA) and an affinity constant Ka of 6.53±2.01×106,M,1 were determined. These results compare well with those measured by fluorescence using untagged GSH, suggesting that the attachment of Fc to GSH does not significantly perturb the GSH structure and binding behavior. By contrasting the binding behavior to several compounds that are known to conjugate to different domains of BSA, the voltammetric study confirmed that GSH-Fc binds at subdomain IIA of BSA with high affinity. The versatility of GSH-Fc for studying GSH binding to surface-confined proteins was also demonstrated with the GSH binding to electroinactive Zn-metallothionein (Zn7 -MT) through hydrogen binding at the region between the Zn7 -MT , and , domains. [source] Dynamic coating of SU-8 microfluidic chips with phospholipid disksELECTROPHORESIS, Issue 15 2010Tiina Sikanen Abstract In this work, PEG-stabilized phosphatidylcholine lipid aggregates (disks), mimicking mammalian cell membranes, were introduced as a new biofouling resistant coating for SU-8 polymer microchannels. A rapid and simple method was developed for immobilization of PEGylated phosphatidylcholine disks in microchannels. Microfluidic chips made from SU-8, PDMS, or glass were dynamically coated with the PEGylated disks followed by characterization of their surface chemistry before and after coating. On the basis of the observed changes in EOF and nonspecific protein adsorption, the affinity of the PEGylated disks was shown to be particularly strong toward SU-8. The PEG-lipid coating enabled permanent change in EOF in SU-8 microchannels with an initial value of 4.5×10,8,m2,V,1,s,1, decreasing to 2.1×10,8,m2,V,1,s,1 (immediately after modification), and, eventually, to 1.5×10,8,m2,V,1,s,1 (7 days after modification) for 9,mM sodium borate (pH 10.5) as BGE. As determined by the Wilhelmy plate measurements and microchip-CE analysis of BSA, the PEG-lipid coating also enabled efficient biofouling shield against protein adsorption, similar to that of low amounts of SDS (3.5,mM) or Tween-20 (80,,M) as buffer additives. These results suggest that dynamically attached PEG-lipid aggregates provide stable, biomimicking surface modification that efficiently reduces biofouling on SU-8. [source] Microfluidic devices for electrokinetic sample fractionationELECTROPHORESIS, Issue 15 2010Zhen Wang Abstract We present three generations of microchip-based "in-space" sample fractionators and collectors for use in proteomics. The basic chip design consisted of a single channel for CE separation of analytes that then intersects a fractionation zone feed into multiple high aspect ratio microchannels for fractionation of separated components. Achievements of each generation are discussed in relation to important design criteria. CE-separated samples were electrokinetically driven to multiple collection channels in sequence without cross-contamination under the protection of sheath streams. A 36-channel fractionator demonstrated the efficacy of a high-throughput fractionator with no observed cross-contamination. A mixture of IgG and BSA was used to test the efficiency of the fractionator and collector. CE of the fractionated samples was performed on the same device to verify their purity. Our demonstration proved to be efficient and reproducible in obtaining non-contaminated samples over 15 sample injections. Experimental results were found to be in close agreement with PSpice simulation in terms of flow behavior, contamination control and device performance. The design presented here has a great potential to be integrated in proteomic platforms. [source] Sensitive, label-free protein assay using 1-ethyl-3-methylimidazolium tetrafluoroborate-supported microchip electrophoresis with laser-induced fluorescence detectionELECTROPHORESIS, Issue 9 2008Yuanhong Xu Abstract Based on the dimer,monomer equilibrium movement of the fluorescent dye Pyronin Y (PY), a rapid, simple, highly sensitive, label-free method for protein detection was developed by microchip electrophoresis with LIF detection. PY formed a nonfluorescent dimer induced by the premicellar aggregation of an anionic surfactant, SDS, however, the fluorescence intensity of the system increased dramatically when proteins such as BSA, bovine hemoglobin, cytochrome c, and trypsin were added to the solution due to the transition of dimer to fluorescent monomer. Furthermore, 1-ethyl-3-methylimidazolium tetrafluoroborate (EMImBF4) instead of PBS was applied as running buffers in microchip electrophoresis. Due to the excellent properties of EMImBF4, not only nonspecific protein adsorption was more efficiently suppressed, but also approximately ten-fold higher fluorescence intensity enhancement was obtained than that using PBS. Under the optimal conditions, detection limits for BSA, bovine hemoglobin, cytochrome c, and trypsin were 1.00×10,6, 2×10,6, 7×10,7, and 5×10,7 mg/mL, respectively. Thus, without covalent modification of the protein, a protein assay method with high sensitivity was achieved on microchips. [source] Quantification of carbonylated proteins in rat skeletal muscle mitochondria using capillary sieving electrophoresis with laser-induced fluorescence detectionELECTROPHORESIS, Issue 2 2008Juan Feng Abstract Carbonyl-modified proteins are markers of oxidative damage. Here, we report a new method for detecting and quantifying carbonylated proteins by capillary sieving electrophoresis (CSE) with LIF detection (CSE-LIF). Alexa 488 hydrazide is used for the specific labeling of carbonyls while 3-(2-furoyl) quinoline-2-carboxaldehyde (FQ) is used for protein labeling. BSA subjected to metal-catalyzed oxidation is used to optimize the labeling reactions, confirm the separation power of CSE, and characterize the response of the LIF detector. The method is capable of detecting femtomole (fmol) amounts of carbonyls in proteins with molecular masses ranging from 26 to 30,kDa. Using this method, we determined that mitochondrial proteins isolated from skeletal muscle contains 2.1,±,0.1 (average,±,SD; n,=,3) nmol carbonyl/mg protein. The methodology described here should be compatible with the analysis of single cells and needle biopsies taken from oxidative stress animal models. [source] On-line concentration of proteins by SDS-CGE with LIF detectionELECTROPHORESIS, Issue 2 2008Cheng-Ju Yu Abstract We present a simple approach for on-line concentration of SDS-protein complexes by using poly(vinyl alcohol) (PVA) solution in CGE. In comparison to the coated capillary, the presence of EOF in CGE omitted the need to fill the capillaries with polymer solutions prior to the analysis. More importantly, we found that highly reproducible separation of eight proteins by 3.5% PVA was achieved between runs and without the regeneration of high bulk EOF; the RSD of migration times was less than 0.7%. To further improve the concentration sensitivity, neutral PVA was introduced into the capillary with the help of EOF to act as sieving matrix. The occurrence of stacking at the boundary between the PVA and the sample zone is mainly due to the retardation of proteins by PVA. As a result, the LODs at an S/N of 3 for SDS,protein complexes are of the order of sub-nM to several nM. For example, the LOD for BSA is 0.78 nM, which is a 91-fold sensitivity enhancement over the normal injection. In addition, our stacking method has been applied to the analyses of proteins in Escherichia coli cells. The peak for ,-galactosidase (E. coli) was observed after 0.1 ,M ,-galactosidase was spiked into the E. coli samples. [source] Measurement of specific radioactivity in proteins separated by two-dimensional gel electrophoresisELECTROPHORESIS, Issue 5-6 2006Shaobo Zhou Abstract We report a method to quantify the specific radioactivity of proteins that have been separated by 2-DE. Gels are stained with SyproRuby, and protein spots are excised. The SyproRuby dye is extracted from each spot using DMSO, and the fluorescence is quantified automatically using a plate reader. The extracted gel piece is then dissolved in hydrogen peroxide and radioactivity is quantified by liquid scintillation counting. Gentle agitation with DMSO for 24,h was found to extract all the SyproRuby dye from gel fragments. The fluorescence of the extract was linearly related to the amount of BSA loaded onto a series of 1-D gels. When rat muscle samples were run on 2-DE gels, the fluorescence extracted from 54,protein spots showed a good correlation (r = 0.79, p < 0.001) with the corresponding spot intensity measured by conventional scanning and image analysis. DMSO extraction was found not to affect the amount of radioactive protein left in the gel. When a series of BSA solutions of known specific radioactivity were run on 2-DE gels, the specific radioactivity measured by the new method showed a good correlation (r = 0.98, p < 0.01, n = 5) with the specific radioactivity measured directly before loading. Reproducibility of the method was measured in a series of 2-DE gels containing proteins from the livers of rats and mice that had been injected with [35S]methionine. Variability tended to increase when the amount of radioactivity in the protein spot was low, but for samples containing at least 10,dpm above background the CV was around 30%, which is comparable to that obtained when measuring protein expression by conventional image analysis of SyproRuby-stained 2-DE gels. Similar results were obtained whether spots were excised manually or using a spot excision robot. This method offers a high-throughput, cost-effective and reliable method of quantifying the specific radioactivity of proteins from metabolic labelling experiments carried out in,vivo, so long as sufficient quantities of radioactive tracer are used. [source] A systematic investigation into the recovery of radioactively labeled proteins from sodium dodecyl sulfate-polyacrylamide gelsELECTROPHORESIS, Issue 1 2004Shaobo Zhou Abstract We report the results of a systematic investigation designed to optimize a method for quantifying radioactivity in proteins in sodium dodecyl sulfate-polyacrylamide gels. The method involves dissolving appropriately sized pieces of gel in hydrogen peroxide and heating to 70°C overnight followed by liquid scintillation counting. H2O2 had no effect on the count rates of [14C]bovine serum albumin (BSA) when counted in a conventional liquid scintillation system, and the count rates remained stable for several days. Temperatures below 70°C resulted in incomplete extraction of radioactivity from gels containing [14C]BSA, but there was also a significant reduction in count rates in samples incubated at 80°C. At 70°C recovery was not affected by the amount of sample loaded onto the gel or by the staining procedure (Coomassie Brilliant Blue or SYPRO Ruby). Recoveries were in the range of 89,94%, and the coefficient of variation for five replicate samples was 5,10%. This method offers a reliable way of measuring the amount of radioactivity in proteins that have been separated by electrophoresis. It may be useful, for example, in quantitative metabolic labeling experiments when it is necessary to know precisely how much tracer has been incorporated into a particular protein. [source] High-efficiency peptide analysis on monolithic multimode capillary columns: Pressure-assisted capillary electrochromatography/capillary electrophoresis coupled to UV and electrospray ionization-mass spectrometryELECTROPHORESIS, Issue 21 2003Alexander R. Ivanov Abstract High-efficiency peptide analysis using multimode pressure-assisted capillary electrochromatography/capillary electrophoresis (pCEC/pCE) monolithic polymeric columns and the separation of model peptide mixtures and protein digests by isocratic and gradient elution under an applied electric field with UV and electrospray ionization-mass spectrometry (ESI-MS) detection is demonstrated. Capillary multipurpose columns were prepared in silanized fused-silica capillaries of 50, 75, and 100 ,m inner diameters by thermally induced in situ copolymerization of methacrylic monomers in the presence of n -propanol and formamide as porogens and azobisisobutyronitrile as initiator. N -Ethylbutylamine was used to modify the chromatographic surface of the monolith from neutral to cationic. Monolithic columns were termed as multipurpose or multimode columns because they showed mixed modes of separation mechanisms under different conditions. Anion-exchange separation ability in the liquid chromatography (LC) mode can be determined by the cationic chromatographic surface of the monolith. At acidic pH and high voltage across the column, the monolithic stationary phase provided conditions for predominantly capillary electrophoretic migration of peptides. At basic pH and electric field across the column, enhanced chromatographic retention of peptides on monolithic capillary column made CEC mechanisms of migration responsible for separation. The role of pressure, ionic strength, pH, and organic content of the mobile phase on chromatographic performance was investigated. High efficiencies (exceeding 300,000 plates/m) of the monolithic columns for peptide separations are shown using volatile and nonvolatile, acidic and basic buffers. Good reproducibility and robustness of isocratic and gradient elution pressure-assisted CEC/CE separations were achieved for both UV and ESI-MS detection. Manipulation of the electric field and gradient conditions allowed high-throughput analysis of complex peptide mixtures. A simple design of sheathless electrospray emitter provided effective and robust low dead volume interfacing of monolithic multimode columns with ESI-MS. Gradient elution pressure-assisted mixed-mode separation CE/CEC-ESI-MS mass fingerprinting and data-dependent pCE/pCEC-ESI-MS/MS analysis of a bovine serum albumin (BSA) tryptic digest in less than 5 min yielding high sequence coverage (73%) demonstrated the potential of the method. [source] Surface grafting of glycidyl methacrylate on silica gel and polyethylene beadsELECTROPHORESIS, Issue 18 2003Seong-Ho Choi Abstract Surface grafting of glycidyl methacrylate (GMA) on silica gel and a polyethylene bead was performed by radical polymerization and radiation-induced polymerization, respectively, in order to improve softness. Subsequently, diethylene triamine (DETA), triethylene tetraamine (TETA), and iminodiacetic acid (IDA) were introduced to the grafted GMA for use as affinity columns. The efficiency of the affinity column was investigated by use of bovine serum albumin (BSA) and hemoglobin (Hb) as model proteins. The affinity degree of BSA was higher than Hb for the DETA and TETA column, whereas the affinity degree of Hb was higher than BSA for the IDA column supported by silica gel. The affinity degree of BSA was higher than Hb for the DETA and TTA column supported by polyethylene (PE) beads. [source] Lanthanide-Based Conjugates as Polyvalent Probes for Biological LabelingEUROPEAN JOURNAL OF INORGANIC CHEMISTRY, Issue 18 2008Stéphanie Claudel-Gillet Abstract A series of lanthanide complexes of [LnL(H2O)] composition, suitable for biological labeling has been studied, in which L is a strongly chelating ligand containing chromophoric bipyridylcarboxylate units and Ln = Sm, Eu, Gd, Tb, and Dy. For the Gd complex, a combined 17O NMR and 1H NMRD study has been performed. The water exchange rate obtained, kex298 = (5.2,±,0.6),×,106 s,1, is slightly higher than those for [Gd(dota)(H2O)], or [Gd(dtpa)(H2O)]2,. Transformation of the uncoordinated carboxylate function of the ligand into an activated ester ensures covalent linking of the complex to bovine serum albumine (BSA). The relaxivity properties of the Gd complex labeled on BSA revealed a limited increase of both longitudinal and transversal relaxivities. This can be related to the partial replacement of the inner-sphere water molecules by coordinating functions of the protein. Additionally, the Sm and Dy complexes are described and chemically characterized. Their photophysical properties were investigated by means of absorption, steady-state and time-resolved spectroscopy, evidencing efficient photosensitization of the lanthanide emission by ligand excitation (antenna effect). Luminescence lifetime measurements confirmed the presence of a water molecule in the first coordination sphere that partly explained the relatively poor luminescence properties of the Dy and Sm complexes in aqueous solutions. The spectroscopic properties of the series of complexes are questioned in terms of time-resolved acquisition techniques. Finally, their availability for use in time-resolved luminescence microscopy is demonstrated by staining experiments of rat brain slices, where the complex showed enhanced localization in some hydrophilic regions of the blood,brain barrier (BBB).(© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008) [source] Spectroscopic investigation of the function of aqueous 2-hydroxyethylmethacrylate/glutaraldehyde solution as a dentin desensitizerEUROPEAN JOURNAL OF ORAL SCIENCES, Issue 4 2006Chuangye Qin Fourier-transform (FT)-Raman and -infrared (IR) spectroscopy were employed to investigate the function of the aqueous 2-hydroxyethylmethacrylate/glutaraldehyde solution (Gluma) as a desensitizer. 2-Hydroxyethylmethacrylate (HEMA), glutaraldehyde (GA), and the mixture of HEMA/GA (i.e. Gluma) were used to interact with dentin, collagen, hydroxyapatite (HAP), and bovine serum albumin (BSA) individually. All the interactions were monitored by an FT-Raman spectrometer. FT-IR spectroscopy was also used in this study. The results show that HEMA could be absorbed by dentin and collagen; GA could cross-link collagen and BSA; and when BSA was added to Gluma, polymerization of HEMA occurred. The results suggest that Gluma acts as a desensitizer whereby, first, GA reacts with part of the serum albumin in dentinal fluid, which induces a precipitation of serum albumin, then, second, a reaction of GA with serum albumin induces polymerization of HEMA. The function of Gluma as a desensitizer to block dentinal tubules occurs via these two reactions. [source] Bovine Serum Albumin and Lysozyme Adsorption on Calcium Phosphate ParticlesADVANCED ENGINEERING MATERIALS, Issue 1-2 2010Berit Mueller Two model proteins that are oppositely charged at neutral pH , bovine serum albumin (BSA) and lysozyme, with acidic and alkaline isoelectric points, respectively , are used to investigate the protein adsorption behaviour of hydroxyapatite and beta-tricalcium phosphate (, -TCP) particles. Both calcium phosphate based particles are highly relevant for the fabrication of bioactive and resorbable bone implants. The investigations are carried out by combining zeta potential and Vis spectroscopy measurements. The changes of zeta potential and isoelectric point are determined as a function of added protein. Both proteins form a monolayer on , -TCP, while on hydroxyapatite only semi-monolayers were measured. For BSA, a side-on adsorption mode is suggested, whereas end-on adsorption appears to be most likely for lysozyme. The zeta potential curves as a function of adsorbed protein show that plateaus of the protein amounts adsorbed increase with charge saturation. In addition, the spatial charge distribution of both proteins is modelled to get a further understanding of the initial adsorption orientation of the biomolecules, supporting the findings from the experimental data. The reported findings can be transferred to the adsorption behaviour of a variety of proteins on calcium phosphate surfaces and are helpful for the fabrication of bone-analogous calcium phosphate/protein nanocomposites. [source] Synthesis and Characterization of Thrombin Conjugated ,-Fe2O3 Magnetic Nanoparticles for HemostasisADVANCED ENGINEERING MATERIALS, Issue 12 2009Ofra Ziv Abstract Thrombin is the final protease produced in the clotting pathways. Thrombin has been used in the clinic more than six decades for topical hemostasis and wound management. In human plasma the half-life of thrombin is shorter than 15 seconds due to close control by inhibitors. In order to stabilize thrombin, this enzyme was conjugated covalently and physically to ,-Fe2O3 magnetic nanoparticles. The physical conjugation was accomplished through adsorption of thrombin to BSA coating on the nanoparticles. The coagulant activity of the covalently bound thrombin was significantly lower than that of the physically adsorbed thrombin. Leakage of the physically bound thrombin into PBS containing 4% HSA was negligible. The physical conjugation of thrombin onto the nanoparticles stabilized the thrombin against its major inhibitor antithrombin III and improved its storage stability. At optimal CaCl2 concentration, the clotting time by the bound thrombin is shorter than that of the free enzyme. This novel conjugated thrombin may be an efficient candidate for topical hemostasis and wound healing. [source] Conversion of vitamin D3 to 1,,25-dihydroxyvitamin D3 in human skin equivalentsEXPERIMENTAL DERMATOLOGY, Issue 2 2000B. Lehmann Abstract: These results demonstrate for the first time that human keratinocytes under in vivo -like conditions have the capacity of the enzymatic hydroxylation of vitamin D3 to hormonally active calcitriol (1,,25(OH)2D3). Supplementation of the culture medium with bovine serum albumin (BSA) up to 1.5% (w/v) amplifies the conversion of vitamin D3 to 1,,25(OH)2D3. The maximum turnover rate of this reaction at 780 nM vitamin D3 in presence of 1.0% (w/v) BSA amounts to approximately 3 pmol 1,,25(OH)2D3 per 106 cells after 6 h of incubation. The hydroxylation of vitamin D3 to 1,,25(OH)2D3 is inhibited by the P-450 oxidase inhibitor ketoconazole. The generation of 1,,25(OH)2D3 from vitamin D3 has an apparent Michaelis constant (Km) of 2.3×10,6 M. The intrinsic conversion of vitamin D3 to biologically active 1,,25(OH)2D3 may be of importance for the regulation of proliferation and differentiation of keratinocytes. [source] Disulfide bond formation through Cys186 facilitates functionally relevant dimerization of trimeric hyaluronan-binding protein 1 (HABP1)/p32/gC1qRFEBS JOURNAL, Issue 1 2002Babal Kant Jha Hyaluronan-binding protein 1 (HABP1), a ubiquitous multifunctional protein, interacts with hyaluronan, globular head of complement component 1q (gC1q), and clustered mannose and has been shown to be involved in cell signalling. In vitro, this recombinant protein isolated from human fibroblast exists in different oligomeric forms, as is evident from the results of various independent techniques in near-physiological conditions. As shown by size-exclusion chromatography under various conditions and glutaraldehyde cross-linking, HABP1 exists as a noncovalently associated trimer in equilibrium with a small fraction of a covalently linked dimer of trimers, i.e. a hexamer. The formation of a covalently-linked hexamer of HABP1 through Cys186 as a dimer of trimers is achieved by thiol group oxidation, which can be blocked by modification of Cys186. The gradual structural transition caused by cysteine-mediated disulfide linkage is evident as the fluorescence intensity increases with increasing Hg2+ concentration until all the HABP1 trimer is converted into hexamer. In order to understand the functional implication of these transitions, we examined the affinity of the hexamer for different ligands. The hexamer shows enhanced affinity for hyaluronan, gC1q, and mannosylated BSA compared with the trimeric form. Our data, analyzed with reference to the HABP1/p32 crystal structure, suggest that the oligomerization state and the compactness of its structure are factors that regulate its function. [source] A folding variant of human ,-lactalbumin induces mitochondrial permeability transition in isolated mitochondriaFEBS JOURNAL, Issue 1 2001Camilla Köhler A human milk fraction containing multimeric ,-lactalbumin (MAL) is able to kill cells via apoptosis. MAL is a protein complex of a folding variant of ,-lactalbumin and lipids. Previous results have shown that upon treatment of transformed cells, MAL localizes to the mitochondria and cytochrome c is released into the cytosol. This is followed by activation of the caspase cascade. In this study, we further investigated the involvement of mitochondria in apoptosis induced by the folding variant of ,-lactalbumin. Addition of MAL to isolated rat liver mitochondria induced a loss of the mitochondrial membrane potential (,,m), mitochondrial swelling and the release of cytochrome c. These changes were Ca2+ -dependent and were prevented by cyclosporin A, an inhibitor of mitochondrial permeability transition. MAL also increased the rate of state 4 respiration in isolated mitochondria by exerting an uncoupling effect. This effect was due to the presence of fatty acids in the MAL complex because it was abolished completely by BSA. BSA delayed, but failed to prevent, mitochondrial swelling as well as dissipation of ,,m, indicating that the fatty acid content of MAL facilitated, rather than caused, these effects. Similar results were obtained with HAMLET (human ,-lactalbumin made lethal to tumour cells), which is native ,-lactalbumin converted in vitro to the apoptosis-inducing folding variant of the protein in complex with oleic acid. Our findings demonstrate that a folding variant of ,-lactalbumin induces mitochondrial permeability transition with subsequent cytochrome c release, which in transformed cells may lead to activation of the caspase cascade and apoptotic death. [source] A spectroscopic study of the reaction of NAMI, a novel ruthenium(III)anti-neoplastic complex, with bovine serum albuminFEBS JOURNAL, Issue 4 2000Luigi Messori The reaction of Na[transRuCl4Me2SO(Im)] (NAMI; where Im is imidazole), a novel anti-neoplastic ruthenium(III) complex, with BSA, was studied in detail by various physico-chemical techniques. It is shown that NAMI, following chloride hydrolysis, binds bovine serum albumin tightly; spectrophotometric and atomic absorption data point out that up to five ruthenium ions are bound per albumin molecule when BSA is incubated for 24 h with an eightfold excess of NAMI. CD and electronic absorption results show that the various ruthenium centers bound to albumin exhibit well distinct spectroscopic features. The first ruthenium equivalent produces a characteristic positive CD band at 415 nm whereas the following NAMI equivalents produce less specific and less marked spectral effects. At high NAMI/BSA molar ratios a broad negative CD band develops at 590 nm. Evidence is provided that the bound ruthenium centers remain in the oxidation state +3. By analogy with the case of transferrins it is proposed that the BSA-bound ruthenium ions are ligated to surface histidines of the protein; results from chemical modification experiments with diethylpyrocarbonate seem to favor this view. Spectral patterns similar to those shown by NAMI are observed when BSA is reacted with two strictly related ruthenium(III) complexes Na[transRuCl4(Me2SO)2] and H(Im)[transRuCl4(Im)2] (ICR), implying a similar mechanism of interaction in all cases. It is suggested that the described NAMI-BSA adducts may form in vivo and may be relevant for the biological properties of this complex; alternatively NAMI/BSA adducts may be tested as specific carriers of the ruthenium complex to cancer cells. Implications of these findings for the mechanism of action of NAMI and of related ruthenium(III) complexes are discussed. [source] Glucose-Responsive Bioinorganic Nanohybrid Membrane for Self-Regulated Insulin ReleaseADVANCED FUNCTIONAL MATERIALS, Issue 9 2010Claudia R. Gordijo Abstract A bioinorganic nanohybrid glucose-responsive membrane is developed for self-regulated insulin delivery analogous to a healthy human pancreas. The application of MnO2 nanoparticles as a multifunctional component in a glucose-responsive, protein-based membrane with embedded pH-responsive hydrogel nanoparticles is proposed. The bio-nanohybrid membrane is prepared by crosslinking bovine serum albumin (BSA),MnO2 nanoparticle conjugates with glucose oxidase and catalase in the presence of poly(N -isopropyl acrylamide- co -methacrylic acid) nanoparticles. The preparation and performance of this new nanocomposite material for a glucose-responsive insulin release system is presented. The activity and stability of immobilized glucose oxidase and the morphology and mechanical properties of the membrane are investigated. The enzymatic activity is well preserved in the membranes. The use of MnO2 nanoparticles not only reinforces the mechanical strength and the porous structure of the BSA-based membrane, but enhances the long-term stability of the enzymes. The in vitro release of insulin across the membrane is modulated by changes in glucose concentration mimicking possible fluctuations of blood-glucose level in diabetic patients. A four-fold increase in insulin permeation is observed when the glucose concentration is increased from normal to hyperglycemic levels, which returns to the baseline level when the glucose concentration is reduced to a normal level. [source] Stabilization of PbS Nanocrystals by Bovine Serum Albumin in its Native and Denatured StatesADVANCED FUNCTIONAL MATERIALS, Issue 9 2009Mandeep Singh Bakshi Abstract PbS nanocrystals (NCs) are synthesized in aqueous phase within a temperature range of 40,80,°C in the presence of native and denatured states of bovine serum albumen (BSA) as the capping/stabilizing agent. The NCs are characterized with the help of field-emission scanning electron microscopy, high-resolution transmission electron microscopy, X-ray diffraction, and energy-dispersive X-ray analysis. At 40,°C, large ball-shaped NCs (145,±,37,nm) with small surface protrusions are formed when 1,×,10,4,g mL,1 BSA is used. As the reaction temperature is increased towards 80,°C, the size of NCs decreases and they acquire somewhat cubic geometries (49.1,±,7.0,nm) due to a change in the capping behavior of BSA between its native and denatured states. The native and denatured states of BSA are simultaneously studied by fluorescence spectroscopy using tryptophan emission, and pH measurements with respect to time and temperature. Gel electrophoresis is used to determine the polarity of the BSA capped NCs. Only the small sized NCs conjugated with relatively larger amounts of BSA show a displacement towards the positively charged electrode in comparison to larger NCs with lower amounts of BSA capping. It was concluded that the denatured state of BSA is more effective in controlling the crystal growth of PbS than its native state especially in the low concentration range. [source] Direct Spectroscopic Evidence that the Photochemical Outcome of Flutamide in a Protein Environment is Tuned by Modification of the Molecular Geometry: A Comparison with the Photobehavior in Cyclodextrin and VesiclesHELVETICA CHIMICA ACTA, Issue 2 2003Salvatore Sortino The photoreactivity of the phototoxic anticancer drug flutamide (FM) in the presence of bovine serum albumin (BSA) has been investigated. The presence of BSA induces a remarkable modification of the photochemical outcome of the drug with respect to that observed in aqueous solution. Induced circular dichroism (ICD) measurements combined with theoretical calculations provide strong evidence that the new photochemical scenario is tuned by changes of the molecular geometry of FM when incorporated in the protein microenvironment. This behavior presents close analogies to that found in the presence of either cyclodextrin or phospholipid vesicles, chosen as models for biological systems, and delineates a quite general photochemical picture that can be useful for a more appropriate understanding of the adverse phototoxic effects induced by this drug. [source] Self-Assembled Organic,Inorganic Hybrid Elastic Crystal via Biomimetic MineralizationADVANCED MATERIALS, Issue 33 2010Halei Zhai Organic,inorganic hybrid rhombs with regular shape and a lamellar superstructure are biomimetically fabricated by the cooperative self-assembly of bovine serum albumin (BSA), sodium bis-2-ethylhexyl sulfosuccinate (AOT), and calcium phosphate (see figure). Although crystalline calcium phosphate is the main component, the hybrid crystals are elastic and they can be flexible under external stresses. [source] Inactivation of root canal medicaments by dentine, hydroxylapatite and bovine serum albuminINTERNATIONAL ENDODONTIC JOURNAL, Issue 3 2001I. Portenier Abstract Aim This study examined and compared the inhibition of the antibacterial effect of saturated calcium hydroxide solution, chlorhexidine acetate and iodine potassium iodide by dentine, hydroxylapatite and bovine serum albumin. MethodologyEnterococcus faecalis strain A197A prepared to a suspension of 3 × 108 cells per ml in 0.5% peptone water was used. Fifty µL of saturated calcium hydroxide solution, 0.05% chlorhexidine acetate or 0.2/0.4% iodine potassium iodide were incubated at 37 °C with 28 mg dentine powder (DP), hydroxylapatite (HA) or bovine serum albumin (BSA) in 50 µL water for 1 h before adding 50 µL of the bacterial suspension. Samples for bacterial culturing were taken from the suspension 1 and 24 h after adding the bacteria. In further experiments, the amount of dentine was stepwise reduced from 28 mg 150 µL,1 to 2.8 mg 150 µL,1. Results Calcium hydroxide was totally inactivated by the presence of 28 mg of DP, HA or BSA. Chlorhexidine (0.05%) was strongly inhibited by BSA and slowed down by dentine. However, HA had little or no inhibitory effect on chlorhexidine. The antibacterial effect of 0.2/0.4% iodine potassium iodide on E. faecalis was totally inhibited by dentine (28 mg), but was practically unaffected by HA or BSA. A stepwise reduction of dentine from 28 mg 150 µL,1 to 2.8 mg 150 µL,1 was followed by a similar reduction of the inhibition of the antibacterial activity of chlorhexidine. Iodine potassium iodide was not inhibited at all with dentine amounts less than 28 mg. However, the effect of saturated calcium hydroxide solution was totally eliminated by dentine, in all four concentrations. Conclusion Inhibition by dentine of the antibacterial activity of calcium hydroxide, chlorhexidine and iodine potassium iodide occurs by different mechanisms. Different components of dentine may be responsible for the inhibition of these three medicaments. Calcium hydroxide was particularly sensitive to inhibition by both inorganic and organic compounds. [source] Suppression of hepatocellular carcinoma by transplantation of ex-vivo immune-modulated NKT lymphocytesINTERNATIONAL JOURNAL OF CANCER, Issue 3 2005Maya Margalit Abstract NKT cells are a regulatory subset of T lymphocytes with immune modulatory effects and an important role in anti-tumor immunity. The feasibility of "ex-vivo education" of NKT cells has recently been demonstrated. To evaluate the anti-tumor effect of ex-vivo immune-modulated NKT lymphocytes in a murine model of hepatocellular carcinoma. Athymic Balb/C mice were sublethally irradiated and transplanted with human Hep3B HCC. NKT cells prepared from immunocompetent Balb/C mice were pulsed ex vivo with HCC-derived antigens (Group A), Hep3B cells (group B) or BSA (group C), and adoptively transferred into HCC harboring mice (1 × 06 NKT cells per mouse). Group D mice did not undergo NKT cell transplantation. Group E mice were transplanted with 1 × 106 NKT cells from HBV-immunized donors. Mice were followed for tumor size and weight. To determine the mechanism of the anti-tumor effect, intrasplenic lymphocyte populations were analyzed by FACS for NKT, CD4+ and CD8+ lymphocyte subpopulations; STAT 1, 4 and 6 expression in splenocytes was assessed by Western blot, and serum cytokine levels were measured by ELISA. Adoptive transfer of NKT cells pulsed with HCC-derived antigens (group A) and NKT cells from immunized donors (group E) resulted in complete disappearance of tumors within 4 weeks and attenuated weight loss (6.5% and 7% in groups A and E, respectively). In contrast, mice in groups B, C, and D developed large, necrotic tumors and severe weight loss (21%, 17% and 23% weight loss in groups B, C, and D, respectively). NKT/CD4 and CD8/CD4 ratios were significantly increased in groups A and E (12.3 and 17.6 in groups A and D, respectively, compared to 6.4, 4.8 and 5.6 in groups B, C and D, respectively, for the NKT/CD4 ratio; 41 and 19.8 in groups A and E, respectively, compared to 6.5, 11.8 and 3.2 in groups B, C, and D, respectively, for the CD8/CD4 ratio). Expression of the transcription factor STAT4 was evident in group A, but not in groups B-D. Serum IFN,, IL12 and IL4 levels were increased in groups A and E. Adoptive transfer of NKT lymphocytes exposed ex vivo by HCC-derived antigens loaded on dendritic cells and NKT cells from immunized donors led to suppression of HCC in mice. NKT-mediated anti-tumor activity was associated increased NKT and CD8+ T lymphocyte numbers, increased expression of STAT4, a marker for IL-12 activity and elevated serum levels of the proinflammatory cytokines IFN, and IL12, and of IL4. Ex-vivo modulation of NKT lymphocytes holds promise as a novel mode of immune therapy for HCC. © 2005 Wiley-Liss, Inc. [source] Characteristics of skin aging in Korean men and womenINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 1 2005J. H. Chung Introduction Korea is located between Japan and Mainland China. The people of these three countries have similar appearances and it is difficult to differentiate between them. Although the population of Asia is more than half of the total population of the Earth, the inherent characteristics of Asian skin have not been well investigated. Commercial markets for cosmetics and drugs for photoaged skin are rapidly expanding in many Asian countries. Therefore, many investigators in the field of dermatology and cosmetology have become interested in brown Asian skin. Clinical characteristics of skin aging and photoaging in Asians Skin aging can be divided into two basic processes: intrinsic aging and photoaging [1]. Intrinsic aging is characterized by smooth, dry, pale, and finely wrinkled skin, whereas photoaging, which indicates premature skin aging in chronically photodamaged skin, is characterized by severe wrinkling and irregular pigmentation. The pattern of wrinkling in Asians seems to differ from that in Caucasians. Asians have coarser, thicker and deep wrinkles, particularly in the forehead, perioral and Crow's foot areas. In contrast, Caucasians usually have relatively fine cheek and Crow's foot wrinkles. The reasons for these differences are not known and need further investigation. There are racial, ethnic and genetic differences, and differences of skin structure and function, between the brown skin of Asians and the white skin of Caucasians. As Asian skin is more pigmented, acute and chronic cutaneous responses to UV irradiation differ from those in white skin. Many people believe, based on clinical impressions, that the main process of photoaging in Asians involves pigmentary changes, rather than wrinkling. However, no study has been performed to confirm this belief. Risk factors for skin wrinkles and their relative risks in Korean skin [2] Various factors such as age, sun-exposure, and smoking are known to be important risk factors for wrinkles. However, the relative risks of each factor on wrinkles in the brown skin of Asians have not been investigated, and they could differ from those in Caucasians. An evaluation system for skin wrinkling is necessary for Asian skin [3]. Thus, we developed an eight-point photographic scale for assessing wrinkles in both Korean genders [2]. This scale can probably be applied to the populations of other Asian countries, at least to the Japanese and Chinese. The pattern of wrinkles in both genders appears to be similar. Age Age is an important risk factor for wrinkling in Asians, as in Caucasians. Korean subjects in their 60s showed a 12-fold increased risk of wrinkling, while subjects in their 70s have a 56-fold increased risk compared with young age group. UV light It is well known that the UV component in sunlight can cause and accelerate photoaging. The pigmented skin of Asian may better protect skin from acute and chronic UV damage. However, we found a strong association between sun-exposure and the development of wrinkling in Koreans. It was found that sun exposure of more than 5 h per day was associated with a 4.8-fold increased risk in wrinkling versus less than 2 h of sun-exposure in Koreans. Estrogen deficiency Korean females have more wrinkles than men, after controlling for age, sun exposure, and smoking, it was found that they have a 3.6-fold increased risk of developing wrinkles than their male counterparts [2]. It has also been reported, that the relative risk for wrinkling in women is higher than in men as for in white Caucasians [4]. The reason why women show more wrinkles remains to be determined. It is possible that a reduction in skin collagen because of estrogen deficiency in postmenopausal woman may aggravate wrinkling severity. Korean women with more than 10 years since menopause showed a 3.9-fold higher risk of wrinkling than the women 5 years of beyond menopause [5]. We demonstrated that women with a history of HRT have a significantly lower risk, more specifically, one fifth of the risk of facial wrinkling relative to those who had no history of HRT. Interestingly, we found that wrinkle severity significantly increased with an increasing number of full term pregnancies. The relative risk for severe wrinkling is increased by approximately 1.8-fold per full term pregnancy. Smoking It is known that smoking causes skin wrinkling in Caucasians, and that it plays no role in Blacks [6, 7]. Koreans with have a smoking history of more than 30 pack years showed a more than 2.8-fold increased risk of wrinkles [2]. The relative risks of wrinkles associated with a 30,50 pack-years history of smoking were 2.8- and 5.5-fold, respectively. Dyspigmentation in Asian skin To follow pigmentary changes, six photographic standards for both genders were developed for Korean skin, to produce a 6-point scale [2, 8]. Hyperpigmented spots, mostly lentigines, were prominent among women, while seborrheic keratosis tended to be more prominent in men. Seborrheic keratosis in Korean men Seborrheic keratoses (SKs) are benign cutaneous tumors. They have diverse clinical and histopathological appearances and are very common in the elderly (over 50 years old). The etiology of SKs is not well understood, although patients with a great number of lesionsshow a familial trait with an autosomal dominant pattern, and human papilloma virus has been suggested as possible cause because of verrucous appearance of the lesions. Exposure to sunlight has been suggested to be a risk factor for SKs. However, there is still some debate in terms of the role of sunlight. Recently, we have investigated the clinical characteristics of SKs and relationship between SKs and sunlight exposure in Korean males [9]. The prevalence of SKs in Koreans increases with age; it rose from 78.9% at 40 years, to 93.9% at 50 years and 98.7% in those over 60 years. Exposed areas, i.e. the face, neck and dorsum of the hands, demonstrate a significant increase in the prevalence of SKs by decade, whereas partly exposed areas, although SKs tended to increase in prevalence with age, this trend was not significant. When the estimated body surface area (BSA) is taken into account, the number of SKs on both the face and dorsum of the hands (0.51 ± 0.08 per 1% BSA) was over-represented compared with the trunk. SKs were also concentrated on the neck (0.38 ± 0.07 per 1% BSA) and in the V-area (0.47 ± 0.09 per 1% BSA). Outer forearms also showed 3-fold more SKs per unit area than neighboring arms and inner forearms, which are classified as partly exposed area (0.09 ± 0.02, 0.03 ± 0.01, respectively). The total area covered by SKs on exposed area also became significantly larger with aging than on intermittently exposed areas. These results indicate that exposure to sunlight might be related to SK growth. Our results indicated that excessive sun exposure is an independent risk factor of SKs. After controlling for age, smoking, and skin type, subjects with a sun exposure history of more than 6 hours per day showed a 2.28-fold increased risk of having severe SKs (n , 6) compared with those exposed for less that 3 h per day. These findings indicated that sun-exposure may play an important role in SK development. In summary, SKs are very common in Korean males and represent one of the major pigmentary problems. SKs concentrate on exposed skin, especially on the face and dorsum of the hands. Both age and lifetime cumulative sunlight exposure are important contributing factors and may work in a synergistic manner. Conclusion Many people tend to believe that wrinkles are not a prominent feature of Asian photoaged skin, and that dyspigmentation is a major manifestation in Asian skin. Contrary to this impression, wrinkling is also a major problem in the photoaged skin of Asians, and Korean people showing severe pigmentary changes usually tend to have severe wrinkles. In conclusion, the wrinkling patterns and pigmentary changes of photoaged skin in East Asians differ from those of Caucasians, and the relative risks of aggravating factors may be different from those of Caucasian skin. References 1.,Gilchrest, B.A. Skin aging and photoaging: an overview. J. Am. Acad. Dermatol. 21, 610,613 (1989). 2.,Chung, J.H. et al. Cutaneous photodamage in Koreans: influence of sex, sun exposure, smoking, and skin color. Arch. Dermatol. 137, 1043,1051 (2001). 3.,Griffiths, C.E. et al. A photonumeric scale for the assessment of cutaneous photodamage. Arch. Dermatol. 128, 347,351 (1992). 4.,Ernster, V.L. et al. Facial wrinkling in men and women, by smoking status. Am. J. Public Health. 85, 78,82 (1995). 5.,Youn, C.S. et al. Effect of pregnancy and menopause on facial wrinkling in women. Acta Derm. Venereol. 83, 419,424 (2003). 6.,Kadunce, D.P. et al. Cigarette smoking: risk factor for premature facial wrinkling. Ann. Intern. Med. 114, 840,844 (1991). 7.,Allen, H.B., Johnson, B.L. and Diamond, S.M. Smoker's wrinkles? JAMA. 225, 1067,1069 (1973). 8.,Chung, J.H. Photoaging in Asians. Photodermatol. Photoimmunol. Photomed. 19, 109,121 (2003). 9.,Kwon, O.S. et al. Seborrheic keratosis in the Korean males: causative role of sunlight. Photodermatol. Photoimmunol. Photomed. 19, 73,80 (2003). [source] A comparison between BSA, PASI, PLASI and SAPASI as measures of disease severity and improvement by therapy in patients with psoriasisINTERNATIONAL JOURNAL OF DERMATOLOGY, Issue 10 2008Tilo Henseler PhD Background, This study investigates four measures of disease severity in patients with psoriasis, both before and after therapy. Methods, Data records were analyzed from 33 patients with moderate-to-severe chronic plaque psoriasis who were treated with efalizumab, 1 mg/kg/week subcutaneously, for 12 weeks. Four measures of disease severity were used: body surface area (BSA), psoriasis area and severity index (PASI), psoriasis log-based area and severity index (PLASI) and self-administered PASI (SAPASI). Results, At the end of the 12-week therapy, the mean percent improvement shown by each measure varied considerably, ranging from 48.6% (PASI) to 70.6% (SAPASI). PASI and PLASI were the most comparable (67.3% and 66.5%). These differences were smaller when a dermatologist's opinion about the improvement was taken into account, for example "very good improvement" ranged from mean percent improvement of 63.8% (BSA) to 83.8% (PASI). The correlation between all measures revealed a high level of significance (P, 10,5). Conclusions, Comparing the slopes and intercepts of the regression lines revealed PLASI as the most reliable measure for the severity and therapeutic improvement in patients with moderate-to-severe chronic plaque psoriasis. PLASI proved to be a marginally more accurate than PASI, and much more accurate than SAPASI and BSA. The superiority of PLASI may be a result of the logarithmic scale of the affected skin surface. [source] | |||||||||||||||||||||||||||||||||||||