if they have tuberculosis, they have a better prognosis. The inal decision on whether to start anti-tuberculous therapy empirically should not be based solely on the concentration of serumalbumin, and other information such as compatible symptoms, physical examination and chest radiography should be taken into account. In Figure 1 we present the hazard ratio and 95% conidence intervals for tuberculosis by serumalbumin concentrations using restricted cubic splines (seven knots) and Cox regression. The low cost of the assay and the general availability in health care facilities in resource-poor settings make serumalbumin very interesting from a public health point of view. In particular, it could be useful to improve the sensitivity and speciicity of the four-symptom screening strategy recommended by the World Health Organization for intensiied case inding and isoniazid preventive therapy. However, we agree with Professor Wiwanitkit in that our study has important limitations. It is a retrospective study using routinely collected data in a setting without tuberculosis culture. Only patients who had the serumalbumin measured at the time of antiretroviral therapy eligibility were included. Forty-three percent of patients did not have a measurement of the serumalbumin and were excluded. Prospective studies are needed to conirm our indings.
Coumarin is a benzopyrone which is widely used as an anti-coagulant, anti-oxidant, anti-cancer and also to cure arthritis, herpes, asthma and inflammation. Here, we studied the binding of synthesized coumarin derivatives with human serumalbumin (HSA) at physiological pH 7.2 by using fluorescence spectroscopy, circular dichroism spectroscopy, molecular docking and molecular dynamics simulation studies. By addition of coumarin derivatives to HSA the maximum fluorescence intensity was reduced due to quenching of intrinsic fluorescence upon binding of coumarin derivatives to HSA. The binding constant and free energy were found to be 1.95760.01610 5 M 21 , 27.175 Kcal M 21 for coumarin derivative (CD) enamide; 0.83760.01610 5 M 21 , 26.685 Kcal M 21 for coumarin derivative (CD) enoate, and 0.60660.01610 5 M 21 , 26.49 Kcal M 21 for coumarin derivative methylprop (CDM) enamide. The CD spectroscopy showed that the protein secondary structure was partially unfolded upon binding of coumarin derivatives. Further, the molecular docking studies showed that coumarin derivatives were binding to HSA at sub-domain IB with the hydrophobic interactions and also with hydrogen bond interactions. Additionally, the molecular dynamics simulations studies contributed in understanding the stability of protein- drug complex system in the aqueous solution and the conformational changes in HSA upon binding of coumarin derivatives. This study will provide insights into designing of the new inspired coumarin derivatives as therapeutic agents against many life threatening diseases.
Schistosomiasis is an endemic parasite disease and praziquantel is the only drug currently in use to control this disease. Experimental and epidemiological evidence strongly suggests that Microtus fortis (Mf) is a naturally resis- tant vertebrate host of Schistosoma japonicum. In the present study, we found that Mf serumalbumin (Mf-albumin) and the conditioned medium of pcDNA3.1-Mf-albumin caused 46.2% and 38.7% schistosomula death rates in 96 h, respectively, which were significantly higher than that of the negative control (p < 0.05). We also found that mice injected with Mf-albumin had a 43.5% reduction in worm burden and a 48.1% reduction in liver eggs per gram (p < 0.05) in comparison to the control animals. To characterise the mechanisms involved in clearance, schistosomula were incubated with fluorescein isothiocyanate-labelled Mf-albumin and fluorescent enrichment effects were found in the gut lumen of schistosomula after 48 h of incubation. Next, digestive tract excretions from schistosomula were collected and the sensitivity of Mf-albumin to digestive tract excretions was evaluated. The results indicated that schistosomula digestive tract excretions showed indigestibility of Mf-albumin. The death of schistosomula could be partially attributed to the lack of digestion of Mf-albumin by digestive tract excretions during the development of the schistosomula stage. Therefore, these data indicate the potential of Mf-albumin as one of the major selective forces for schistosomiasis.
3. Kandagal PB, Ashoka S, Seetharamappa J, Shaikh SM, Jadegoud Y, Ijare OB. Study of the interaction of an antican- cer drug with human and bovine serumalbumin: spectro- scopic approach. J Pharm Biomed Anal 2006; 41: 393-399. 4. Sun SF, Zhou B, Hou HN, Liu Y, Xiang GY. Studies on the interaction between Oxaprozin-E and bovine serum albu- min by spectroscopic methods. Int J Biol Macromol 2006; 39: 197-200.
The aim of the study is to find out the pre-operative serum level and try to find out the complications associated with hypoalbuminemia and surgical complications. The study was done in KVG Medical College, Sullia. Seventy five patients were selected irrespective of the sex and divided into three groups based on the age. In the present study it is very clear that the serumalbumin reflects the overall outcome of the surgery. As the age progresses the amount of hypoalbuminemia encountered is high. The amount of hospital stay also depends on the protein level. Albumin is no doubt the healer protein of the body. As the albumin level decreases the complications faced by the patients drastically increases.
The nutritional questionnaire used in the JAGES 2010 study asked only about fish/meat consumption. A previous study showed that in younger and older adults dietary protein con- sumption and quantity affects serumalbumin level. In a Japanese study, animal protein intake was related to serumalbumin level, although vegetable protein intake was not. In future studies, questions on detailed food groups, such as eggs, beans, and milk, might need to be added to determine how low-income older adults transition to poor nutrition and health, and how such transitions can be prevented. There might be other bridging factors between low serumalbumin level and socioeconomic status.
Many challenges to the accurate structural prediction of HSA binding exist, including the moderate-to-poor resolution of the serumalbumin structures deposited in the PDB and, in many cases, the poor electron density for crystallographic ligand(s). Analyses of structural data for protein-ligand binding must use high-quality information in order to provide accurate insight; a resolution better than 2.5 A ˚ and a real-space correlation coefficient (RSCC) greater than 0.90 are useful filters when selecting crystallographic data . RSCC provides a goodness- of-fit metric, based on the correlation between the map obtained from the structural model and the map calculated from the experimental data . As of publication, two-thirds of the human HSA structures in the PDB have a resolution worse than 2.5 A ˚ (average is 2.6 A ˚ ), and the ligands present in those structures have RSCC ranging from 0.82–0.95. For this reason, we exercise care in comparing computed docking poses to the reported crystallo- graphic ligand poses [28,29]. Furthermore, since the role of FAs in ligand-HSA binding is not well understood, they have not been included in any published calculations of HSA binding. However, since they can influence small molecule binding to HSA, they are important to a complete picture of the significant structural interactions (Figure S1) [30,31]. Finally, many compounds are capable of binding to both sites, albeit with differing affinities, and although binding is both stereo-selective and dose-dependent, the affinities of each enantiomer are not always reported [32,33].
The binding of chlorpromazine (CPZ) and hemin to bovine serumalbumin was studied by the fluorescence quenching technique. CPZ is a widely used anti-psychotic drug that interacts with blood components, influences bioavailability, and affects function of several biomolecules. Hemin is an important ferric residue of hemo- globin that binds within the hydrophobic region of albumin with high specificity. Quenching of the intrinsic fluorescence of bovine serumalbumin (BSA) was observed by selectively exciting tryp- tophan residues at 290 nm. Emission spectra were recorded in the range from 300 to 450 nm for each quencher addition. Stern-Volmer graphs were plotted, and the quenching constant estimated for BSA solution titrated with hemin at 25ºC was 1.44 (± 0.05) x 10 5 M -1 .
The aim of the present study was to characterize biotin-decorated docetaxel-loaded bovine serumalbumin nanoparticles (DTX-BIO-BSA-NPs) and evaluate their antiproliferative activity in vitro. The particle size of prepared DTX-BIO-BSA-NPs was found to be always lower than 200 nm, with sizes of 166.9, 160.3, 159.0, 176.1 and 184.8 nm and the zeta potential was -29.51, -28.54, -36.54, -36.08 and -27.56 mV after redissolution with water for 0, 1, 2, 4 and 8 hours, respectively. The polydispersity index (PDI) was stable in the range of 0.170 - 0.178. In the in vitro drug-release study, the DTX-BIO-BSA-NPs targeted a human breast cancer cell line MCF-7 effectively. The x-ray diffraction spectrum and DSC curve of DTX-BIO-BSA-NPs suggested that docetaxel was in an amorphous or disordered crystalline phase in DTX-BIO-BSA-NPs. In vitro cytotoxicity results showed that DTX-BIO-BSA-NPs inhibits proliferation of MCF-7, SGC7901, LS-174T and A549 cells in a concentration-dependent manner after exposure to DTX-BIO-BSA-NPs for 48 hours. Taken together, these results indicate that DTX-BIO-BSA-NPs may have potential as an alternative delivery system for parenteral administration of docetaxel.
This study presents a BSA sandwich ELISA with a quantifica- tion range from 1.32–40 ng/mL for quantification of BSA from swab samples (figure 2). The ELISA has comparably sensitivity to other BSA sandwich ELISAs, both commercial assays and assays described in the literature [10,11,12]. The benefits of using the described ELISA are that the setup is simple and all components are commercially available. Regular BSA ELISAs have been developed, but the sandwich ELISAs seem to be more advanta- geous for the detection of BSA in samples containing other proteins . The optimal ELISA setup is the assay that presents the highest difference between the upper concentration limit of the BSA standards (80 ng/mL) and the assay background (0 ng/mL). The background absorbance of our ELISA is slightly higher than that of other assays; however, the level is acceptable as the maximum absorbance is at an equally higher level. We tested different blocking dilutions in addition to PBST, such as rabbit serum and blocking at 90 min. at 37 uC as described by Zhang et al. . Although different attempts to decrease non-specific binding failed, PBST blocking was found to be the optimal blocking agent tested in this study. In some of the ELISA setups (9 and 15) in figure 1, the absorbance of the control without primary antibody (with BSA) is lower than the 0 ng/mL BSA standard. This could indicate that the BSA has a blocking effect in these assay setups. BSA is a widely used as a blocking reagent in other ELISAs [14,15].
The biological properties of other low abundance proteins are yet to be determined, but it is interesting to report that bovine colostrum proteins have beneficial effects on some human pa- thologies, as tumor or neurodegenerative diseases, like Alzheimer’s . Many authors have demonstrated that dietary whey proteins could prevent tumors by increasing glutathione levels in serum and tissues as well as enhancing splenic lymphocyte proliferation, T helper and cyto- toxic T cell activity . Moreover, some researchers suggest that also β-lactoglobulin, α-lactal- bumin, serumalbumin, and lactoferrin could have anticancer potential . Lactoferrin, in particular, inhibits intestinal tumors and perhaps tumors in other organs stimulating apopto- sis, inhibiting angiogenesis and modulating carcinogen metabolizing enzymes . The princi- pal obstacle to the detailed study of low abundance proteins in colostrum is the high number of these proteins. In the last decade, proteomics has been established as a reliable and successful approach for the study of complex biological fluids, representing a powerful tool for the simul- taneous analysis of hundred proteins in complex mixtures.
Initially, we used the Shapiro-Wilk test to investigate the framing of the numeric variables in the Gaussian model of distribution. Mann-Whitney’s U test was used to compare the median values for colloid solution administration with enteral diet, PN time, length of hospitalization, serum sodium levels, and serumalbumin levels. The chi-squared test was used to compare the percentages of newborns who presented anasarca. Simple linear regression tests were used to analyze the relationship between length of hospitaliza- tion, gestational age, birth weight, low serum sodium levels, low serumalbumin levels, PN time, and OTI time. A multiple linear regression model was fitted to assess the association between each outcome (serumalbumin levels, serum sodium levels, and the need for albumin solution infusion) and the independent variables. The adjusted analysis was conducted following a previously formulated hierarchical model in three levels: 1) gestational age; 2) birth weight; and 3) hospitalization time, OTI time, time to first enteral feeding, and time of full enteral feeding. In this model, the variables were controlled for factors in the same level and in the superior levels. P-values #0.20 were adopted in the univariate analysis as necessary to determine which variables to include in the multiple analyses or when there was a ß modification of more than 10% for any variable already in the model. The variables were then entered using the forward method. The significance level was set at p,0.05. Moreover, homoscedasticity was assessed graphically in all regression models. The Stata statistical software package, Version 10.0 (Stata Corp., College Station, TX, USA) was used for all statistical calculations.
Protein solutions. apoHTF (ATF2011-07; >95%), was purchased from AkronQ bio tech; fatty acid and globulin free human serumalbumin (Sigma Aldrich, A3782, +99%), lyophilized powder with a molecular mass of 67 kDa was used. Human apo-transferrin solu- tions were prepared by dissolving the protein in buffer at pH 7.4. The solutions were allowed to stand for at least 1 h to allow equili- bration. During this period they were gently swirled without strong agitation. The concentration of apoHTF solutions were de- termined by measuring the absorbance at 280 nm using an extinc- tion coefficient of 92300m @1 cm @1 ; [31,125,126] and for albumin it was
DEVELOPMENT OF RESTRICTED-ACCESS MEDIA CHROMATOGRAPHIC COLUMNS PROTEIN-IMMOBILIZED AND THEIR EVALUATION FOR THE ANALYSIS OF DRUGS BY DIRECT INJECTION OF HUMAN PLASMA. A series of bovine serumalbumin-immobilized supports have been prepared and used as restricted access media (RAM) columns. Restricted-access supports combine size-exclusion of proteins and other high-molar-mass matrix components with the simultaneous enrichment of low-molar mass analytes. These characteristics were chromatographically evaluated for the columns. The RAM-BSA (Bovine SerumAlbumin) columns showed excellent performance for exclusion of human plasma protein with good retention capacity for a series of acidic, basic, and neutral drugs.
Blood samples were taken by from both plain and EDTA tubes. The latter were used for complete blood count (CBC). Most rou- tine CBC were done with a Cell-Dyn Ruby analyzer (Abbott Diagnostics, USA), while most clinical chemistry parameters (creatinine, blood urea nitrogen and serumalbumin) were measured using a DxC800 Synchron analyzer (Beckman Coul- ter, USA). ESR determinations were performed by the Wintrobe method, whose upper normal limit was 20 mm/h.
Only serumalbumin level was found to have a negative correlation with serum total cholesterol level. Age, sex, type of the glomerular disease, serum creatinine, proteinuria and total triglyceride levels, use of statins, steroid, ACEi or ARB were not correlated with LDL cholesterol levels. This may be regarded as indicating that the treatment of hyperlipidemia in nephrotic syndrome should be directed towards increasing serumalbumin levels which is possible with the treatment of the underlying glomerular disease. These may show that serumalbumin level is a stronger indicator of serum lipid levels than urinary albumin loss in primary glomerular diseases. Furthermore, there may be no need for extra medication to treat hyperlipidemia in the initial period of treatment in those without established cardiovascular risk factors. We showed in the present study the decrease in serum total and LDL cholesterol levels in parallel to proteinuria indirectly due to increased serumalbumin levels (Figure 1 and 2).
During experiments assessing the chronic effect of fatty acids on BMMSCs this media was also supplemented with 4% fatty acid free bovine serumalbumin (BSA) (Equitech-Bio Inc BAH66) or 4% BSA bound to the indicated type and concentration of fatty acid (Palmitate, Sigma P9767; Oleate, Fluka Analytical 60420; Stearate, Sigma S3381). For experiments assessing the acute effect of fatty acids, cells were exposed to the normal media described above until begin- ning measurement of metabolism. More information on this assay and the media used that was supplemented with BSA and fatty acids is provided below in the metabolic rates and fatty acid uptake section. Human BMMSCs from a single donor were purchased from the Texas A&M Health Science Center already characterized. Some of the measurements involved in this characterization included confirmation of ability to undergo adipogenesis and osteogenesis, ex- pression of CD105, CD73, and CD90, and absence of CD45, CD34, and CD14 expression. Human BMMSCs were passaged at 70% confluency with 60 cells seeded per cm 2 .
Introduction: To evaluate the relationship between various biochemical, nutritional and demographic factors with immune response to hepatitis B vaccine in maintenance hemodialysis (HD) patients. Material and Methods: A retro-prospective study was carried out on 68 patients undergoing maintenance hemodialysis .Patients were vaccinated against hepatitis B virus with an intramuscular hepatitis B vaccination schedule, 40 micrograms at 0, 1, and 6 months. We also selected 32 age matched normal healthy persons who had vaccinated against hepatitis B previously to compare the antibody production with HD patients. Results: The value of serum antibody level against hepatitis B surface antigen ( HBs) in hemodialisis patients and healthy persons were 35±55(median=5.5) and 135±71 (median=175) mIU/ml respectively. There was a significant deference between mean serum antibody level against HBs antigen of hemodialysis patients and normal subjects (p<0.001), there were not any significant differences of antibody production against HBs antigen between males and females or diabetic and non diabetics. There were no correlation between serum antibody level against HBs-Ag and serumalbumin and also with body mass index. There were not significant correlation between anti-HBs antibody level and age, amounts of hemodialysis, duration of dialysis, dialysis adequacy, serum ferritin level and serum lipids. There were not also significant correlation between anti-HBs antibody level and serum parathormone, calcium, phosphorus, serum hemoglobin and hematocrit level. Conclusion: In this study, there was not significant correlation between serum antibody level against hepatitis B surface antigen and various nutritional and demographic factors of patients under regular hemodialysis.
ELISA tests were done by adding bovine serumalbumin (BSA) or trypsin in the wells to verify the polyclonal antibodies specificity. Enzymatic digestions assays of hFIX (Octanyne ® ) were done using trypsin (Macarrone et al, 2001). The enzymatic digestion was performed in five defined times of 1, 5, 20 and 60 minutes plus 24. The reaction was stopped by adding 60% acetic acid solution stopped the reaction. The peptide pools