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We present the results of studying and selectiing optimal conditions for the determination of caffeine, catechins and gallic acid in black tea by the method of micellar electrokinetic chromatography. The choice of analytes is determined by the fact that they form the main consumer qualities of tea and, due to their antioxidant properties, have a positive effect on human health. Optimization of the electrophoretic detemination of seven catechins, caffeine and gallic acid in black tea was carried out using the method of ex­ periment design. Optimized conditions — 25 mM phosphate buffer solution with pH 7.4, containing 30 mM SDS and 5% ethyl alcohol — provided a satisfactory resolution of all electrophoretogram peaks, the analysis time was 25 minutes, and the current in the system did not exceed 120 pA. After the assessment of the metrological characteristics of the proposed method the standard deviation of the determina­tion of analytes did not exceed 15%. The samples of black tea from Ceylon, Chinese, Assamese, Indian, Kenyan and Krasnodar regions of growth were analyzed, and an array of data on the contents of the analytes under study was formed. Discriminant analysis was used to develop a model and obtain classifica­tion functions for six tea groups from different regions of the world. Proceeding from the obtained classifi­cation functions, a scatter diagram of canonical values was constructed, which showed that the samples of Krasnodar and Chinese tea were localized from all the studied groups. Indian, Assamese and Ceylon teas formed a single area with close to the Chinese tea group. The correctness of the model was checked and revealed the total predictive force about 92%. It is shown that the content of catechins, gallic acid and caf­feine are suitable markers for the classification of black tea samples from different regions of origin.

The response of β-amylase in early stage germination of wheat seeds toBacillus subtilisQM3 was mainly focused on to elucidate the promotion mechanism ofB. subtilisQM3. The results showed that the changes in apparent activity of amylase and endosperm liquefaction after the strain QM3 treatment were much more obvious than that of the control group; the activity of β-amylase treated with the different concentrations of the strain QM3 increased significantly (P< 0.05) by 4% (107 CFU/mL) and 18.5% (106 CFU/mL) at the germination 6 h. Moreover, after presoaking with α-cyclodextrin, the activity of β-amylase increased significantly (P< 0.05) by 18.5% (107 CFU/mL) and 23.4% (106 CFU/mL) at the same stage of germination; the electrophoretogram of β-amylase isoenzymes showed that there is a growing trend in brightness and width of the band during the early germination from 3 to 6 h of wheat seed treated by the strain QM3 (106 CFU/mL). The increase in activity and isoenzyme expression of β-amylase may be one of the important reasons to promote the germination of wheat seeds after being treated byB. subtilisQM3.

The existing miniature capillary electrophoresis (CE) devices use a tablet or a computer for data processing and analysis, which hinders their portability. In order to solve this problem, a smartphone-based CE device was proposed, which allowed for real portable quantitative analysis. The device integrated the functions of capacitively coupled contactless conductivity detection (C4D) and Bluetooth communication. Furthermore, a Kotlin language-based application with a user-friendly interface was developed. The application could not only control the electrophoresis run in the CE device but also receive the data from the C4D detector in real time, display the electrophoretogram, and process the data. The peak areas could be calculated automatically on the smartphone, and the migration time could be obtained. The size of the developed device was 20 cm×20 cm×15 cm, and its weight was 2 kg. Quaternary ammonium salts (QAs) in disinfectors (dodecyl dimethyl benzyl ammonium bromide (DDBAB) and dodecyl trimethyl ammonium bromide (DTAB)) were used as the analytes to verify the performance of the developed device. The experimental data showed that the linear ranges of DDBAB and DTAB were from 20 to 1000 and from 30 to 1000 μmol/L, respectively. The correlation coefficients (R2) of DDBAB and DTAB were 0.9995 and 0.9989, respectively, indicating good linearity between the peak area and concentration. The limits of detection (LODs) of DDBAB and DTAB were 10 and 13 μmol/L, respectively. The intra-day relative standard deviations (RSDs, n=3) of DDBAB and DTAB were 1.9% and 2.7% respectively, revealing good repeatability. In addition, a mixture of DDBAB and DTAB was tested. Two QAs were separated within 8 min, showing good selectivity. Finally, QAs in a bromo geramine disinfector used in the field were tested to further validate the performance of the designed device. The recoveries of DDBAB and DTAB were 100.5%-101.5% and 96.2%-99.3%, respectively, indicating good accuracy. The developed device has the advantages of good linearity, low LOD, good repeatability, high accuracy, and real portability, and it can be used for the quantitative detection of QAs in disinfectors.

Although serum protein electrophoresis is a diagnostic tool available through many veterinary laboratories, there currently are no reference intervals for protein fractions in healthy common mynahs (Acridotheres tristis). Therefore, electrophoretic patterns of proteins in serum and heparinized plasma of the common mynah were evaluated. Blood specimens were collected from 55 healthy adult common mynahs of unknown age (26 males and 29 females). The serum total protein and protein fractions were measured using the biuret method followed by cellulose acetate electrophoresis (CAE). The serum level of albumin was compared with bromocresol green (BCG) dye-binding and CAE methods. Four protein fractions, including albumin and α, β, and γ globulins, were recorded in the electrophoretogram of serum specimens. Sex appeared to have no significant effect on the measured parameters. The serum BCG albumin fraction was significantly higher than the CAE albumin fraction (P = .01). Also, the comparison of total protein and protein fractions in serum and plasma specimens of 25 of the 55 birds sampled showed that total protein (Cohen index d = 0.66, P = .03), gamma globulin (d = 1.13, P = .00), and total globulin (d = 0.67, P = .00) in plasma samples were significantly higher than those in serum samples. The results of this study provide the specific reference intervals for total protein and protein fractions in common mynahs, which are essential for proper interpretation of laboratory results and also revealed that the albumin measurement by the BCG method yields unreliable results in common mynahs.

Seafood traceability is an important means of ensuring the quality of local seafood. Jellyfish are an important edible resource with a large yield on a global scale. Different species of pickled jellyfish, a traditional Chinese food, cannot be differentiated solely from the appearance of the product due to its special processing methods. In this paper, two species of jellyfish, R. esculentum and S. meleagris, were identified by PCR-RFLP technique. The genetic identification of the jellyfish R. esculentum and S. meleagris was based on PCR amplification of a common 651 bp fragment of the 16S rRNA gene, followed by digestion of four restriction enzymes, Hind III, Hpa I, Xho I and Dra I, respectively. The results showed that the two species of fresh jellyfish could be identified clearly, as could the pickled R. esculentum. To verify this result, PCR-RFLP was applied to authenticate four kinds of commercial pickled jellyfish. The electrophoretogram clearly showed that R. esculentum and S. meleagris were easily and accurately authenticated. This PCR-RFLP system has the potential to become an effective method for identifying the jellyfish products from Liaoning Province of China.

Identification and quantitation of enantiomers is a critical and challenging step in the process of chiral capillary electrophoresis (CE) analysis, especially when the optically pure enantiomers are expensive or commercially unavailable. Herein, a method of CE in combination with circular dichroism (CD) spectroscopy for the identification of enantiomeric peak independent of single enantiomer standard was proposed. By comparing the theoretical CD spectrum of the single enantiomer calculated by time-dependent density functional theory (TDDFT) with the experimental CD spectrum of the enantiomeric mixture, the configuration of the dominant enantiomer in the nonracemic mixture was determined. Considering that the dominant enantiomer showed bigger peak area on the CE electrophoretogram, the enantiomeric peak was easily identified. Three kinds of enantiomers including seven chiral compounds (i.e., tryptophan, tyrosine, phenylalanine, Boc-valine, Boc-leucine, ibuprofen, and naproxen) were used to evaluate the reliability of the method. The concentration of the single enantiomer in the mixture can be further accurately quantified based on the total concentration of the mixture and the peak area ratio of a couple of enantiomers, and the accuracy was assessed by taking ibuprofen as an example. The developed CE-CD method provides an alternative tool for the analysis of nonracemic mixture with good ECD signals.

Background Two-dimensional electrophoresis (2-DE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) are widely used in plant proteomics research. However, these two techniques cannot be simultaneously satisfied by traditional protein extraction methods when investigate cotton leaf proteome. Results Here, we evaluated the efficiency of three different protein extraction methods for 2-DE and LC-MS/MS analyses of total proteins obtained from cotton leaves. The protein yield of the borax/PVPP/phenol (BPP) method (0.14%) was significantly lower than the yields of the trichloroacetic acid/acetone (TCA) precipitation method (1.42%) and optimized TCA combined with BPP (TCA-B) method (0.47%). The BPP method was failed to get a clear 2-DE electrophoretogram. Fifty pairs of protein spots were randomly selected from the 2-DE gels of TCA- and TCA-B-extracted proteins for identification by MALDI TOF/TOF, and the results of 42 pairs were consistent. High-throughput proteomic analysis showed that 6339, 9282 and 9697 unique proteins were identified from the total cotton leaf proteins extracted by the TCA, BPP and TCA-B methods, respectively. Gene Ontology (GO) analysis revealed that the proteins specifically identified by TCA method were primarily distributed in the plasma membrane, while BPP and TCA-B methods specific proteins distributed in the cytosol, indicating the sub-cellular preference of different protein extraction methods. Further, ATP-dependent zinc metalloprotease FTSH 8 could be observed in the 2-DE gels of TCA and TCA-B methods, and could only be detected in the LC-MS/MS results of the BPP and TCA-B methods, showing that TCA-B method might be the optimized choice for both 2-DE and LC-MS/MS. Conclusion Our data provided an improved TCA-B method for protein extraction that is compatible with 2-DE and LC-MS/MS for cotton leaves and similar plant tissues which is rich in polysaccharides and polyphenols. Electronic supplementary material The online version of this article (10.1186/s12864-019-5658-5) contains supplementary material, which is available to authorized users.

Up: A schematic-diagram of POU1F1-gene. Down right: an electrophoretogram of the detected novel pathogenic-variant in comparison with wild-type POU1F1 exon-6 sequence. Down left: Family pedigree of the two-siblings reported.

Introduction: Tuberculosis (TB) is a major health problem in India, so early diagnosis and treatment of Mycobacterium tuberculosis (M.tb.) infection can prevent deaths from this pathogen. The secretion of proteins by M.tb. is important in diagnostic purposes for generation of therapeutic drugs and vaccines candidates for TB. The objective of this study was to identify the protein expression (biomarkers) in TB and Tuberculosis meningitis (TBM) using proteomic approach. Methods: In this study, using Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE), we analyzed the secretory proteins of M.tb. in serum, cerebrospinal fluid (CSF) samples. The identified proteins were determined by Total Lab- 100 Quantity One densitometry software. Results: Our study showed that protein bands expressed in CSF samples reveals the presence of 72kD, 70kD, 44kD, 40kD & 16kD predominantly in TBM patients compared to healthy individuals. The electrophoretogram identified 97kD, 72kD, 44kD, 38kD, 29kD & 16kD predominant proteins in serum samples of TB patients. Conclusion: The detection of secretory proteins in serum and CSF samples of M.tb. in TB and TBM patients gives reliable and early diagnosis of TB and TBM. The secretory proteins can be useful as immunodiagnostic and vaccine targets which can serve as important biomarkers

Currently, there is a lack of effective early screening and detection methods for femoral head necrosis. Current research on most orthopedic diseases focuses on proteomics in the preliminary stage. The recent fluorescence differential in gel electrophoresis (DIGE) has advantages such as a high reproducibility, high sensitivity, high throughput, and high dynamic range. It is currently one of the most widely used quantitative proteomic research means. We conducted this study to investigate the pathogenesis of non-traumatic femoral head necrosis using the fluorescence DIGE to screen non-traumatic femoral head necrosis based on proteomics and provide a theoretical basis for screening possible biomarkers and molecular targeted treatment. The DIGE technique was used to separate the protein. An electrophoretogram was established on the basis of scanning and analysis. Identification and a bioinformatics analysis were conducted for the differential protein. The protein with differential expression of over 2-fold was excavated and ionized by means of substrate assisted laser desorption. The flight time was identified with a mass spectrometer (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, MALDI-TOF/TOF). The formation on sequences, structures and functions of these proteins were obtained through database retrieval. Western blot analysis was used to verify the differential protein expression and the reliability of the DIGE result was verified. DIGE was used to successfully separate 1,500±40 protein spots. There were 252 significant differential protein spots. The Ettan™ Spot Picker automatic work station was used to excavate 49 significant differential protein spots with expression difference over 2-fold. The MALDI-TOF/TOF mass spectrometer was used to identify these differential protein spots. Six proteins were identified in total, which include apolipoprotein A1 (APOA1), fibrous protein original chain, fibrous protein original chain, serum albumin, sulfur-oxygen protein peroxiredoxin 2 (PRDX2) and actin. APOA1 and PRDX2 were subject to western blot analysis detection; results were consistent with the DIGE result. Based on an analysis of the biological information, these proteins may be associated with the incidence and progression of femoral head necrosis.

Genetic Profiling of Methicillin-resistant Staphylococcus aureus in An African Hospital by Multiplex-PCR Shuaibu Suleiman Adeiza, Josiah Ademola Onaolapo, Busayo Olalekan Olayinka Department of Pharmaceutical Microbiology, Faculty of Pharmaceutical sciences, Ahmadu Bello University, Zaria, Kaduna, Nigeria ABSTRACT Objectives: Methicillin resistant Staphylococcus aureus (MRSA) nasal carriage often precedes infection. This work aims to genetically profile MRSA carriage of resistant genes (mecA, blaZ and tetM) and certain virulence genes (nuc, pvl, spa and scn). Methods: Phenotypically confirmed Staphylococcus aureus were Screened for oxacillin resistance and assessed for β-lactamase production. Further confirmation of identity was performed by multiplex PCR of the 16SrRNA, mecA, tetM, pvl, nuc, spa and scn genes. DNA gel fingerprint analysis was carried out and the bands were clustered in a dendrogram. Results: The study analysed 81 Staphylococcus aureus isolates for Methicillin resistance out of which 38 (46.9%) were MRSA. The DNA extraction method was effective for all MRSA isolates evaluated. All phenotypic MRSA isolates (100%) were positive for the S. aureus specific 16SrRNA gene confirming that these isolates were S. aureus strains. The nuc gene was detected in 19/38 (50%) of the MRSA isolates, blaZ gene in 14/38 (36.8%) and tetM in 18/38 (47.4%). Conclusions: The need for improved vigilance to recognize MRSA strains cannot be overemphasized. If MRSA control is not tracked, these strains will increase exponentially. J Microbiol Infect Dis 2020; 10(3):144-153.

Autoimmune haemolytic anaemia is a condition in which autoantibodies present against self erythrocytes which leads to severe haemolysis and loss of haemoglobin. In the present study we compared the electrophoretic band patterns between control and autoimmune haemolytic anaemia and non autoimmune iron deficiency anaemia erythrocyte lysate samples. We observed the presence of a band which was specific to autoimmune haemolytic erythrocyte lysates in the region of 70 Kd which was later identified as albumin and was distinctly absent in control as well as iron deficiency non autoimmune haemolytic samples. In addition, we have noticed that the catalase in the region of 60 Kd was found to be decreased in its band intensity in haemolytic anaemia samples compared to control. The presence of both albumin and catalase in the respective positions in the electrophoretic gel was confirmed by in gel trypsin digestion followed by mass spectrometry. The other erythrocyte proteins visible in the region of 25 Kd were found to have an overall decrease in band intensity in haemolytic anaemia samples compared to control. The specific presence of albumin in autoimmune haemolytic erythrocyte lysate is the main highlight of the work which was not reported till date and further studies could reveal the exact role of albumin in autoimmune haemolytic conditions. Key words: Autoimmune haemolytic anaemia, erythrocytes, electrophoresis, albumin, catalase

T4 polynucleotide kinase (T4 PNK) may catalyze the phosphorylation of 5'-hydroxyl termini in nucleic acids, which play a crucial role in DNA recombination, replication and damage repair. Here, a microchip electrophoresis laser induced fluorescence (MCE-LIF) method based on biochemical reaction was developed for the detection of T4 PNK activity and inhibitors. In this method, the single strand DNA (ssDNA) was hybridized with the 5-carboxyfluorescein (FAM) labeled single strand DNA (ssDNA-FAM) to form FAM labeled double-stranded DNA (dsDNA-FAM). In the presence of T4 PNK and adenosine triphosphate (ATP), T4 PNK catalyzes the transfer of γ-phosphate residues from ATP to the 5-hydroxyl terminal of dsDNA-FAM. The phosphorylated dsDNA-FAM can be gradually hydrolyzed by λexo to produce a FAM labeled single nucleotide fragment. Then the FAM labeled single nucleotide fragment and the unhydrolyzed dsDNA-FAM were separated by MCE, and two electrophoresis peaks appeared in the electrophoretogram. The detection of T4 PNK activity and inhibitors was realized by measuring the peak height of the FAM labeled single nucleotide fragment in electrophoretogram. This assay is very sensitive with a limit of detection of 0.002 U/mL, and it can be further used to screen the T4 PNK inhibitors.

In order to improve the quality of peak characteristics extraction in chromatographic data, an automatic peak fitting method based on wavelet ridges and morphology (WMPF) was proposed. In handling overlapping peaks and weak peaks, WMPF gave solution to deal with the shortcomings of existing peak detection methods. The ridge line of the signal was obtained by wavelet transform of the original chromatogram signal, and the signal was divided into peak areas and non-peak areas. Then, we processed morphological operations with an adaptive window, and combined Gaussian fitting with wavelet ridge information. Finally, the peak characteristics of the signal were extracted. Through the analysis of high-density lipoprotein electrophoretogram data, it was confirmed that the proposed method was effective for baseline separation, noise elimination, peak detection and overlapping peak recognition.

It is of significance to elucidate or understand the intracellular transformation & migration behaviors of heavy metals in specific cells. Herein, we report the fast and efficient separation of cadmium-metallothioneins (Cd-MTs) and Cd2+in cell lysate by a short column capillary electrophoresis (SC-CE), followed by coupling with inductively coupled plasma mass spectrometry (ICP-MS) to facilitate the speciation of intracellular cadmium species. The incorporation of sodium dodecyl sulfate (SDS) in running buffer significantly reduces the peak width of Cd2+from 170 s to 26 s in the electrophoretogram, causing a 5.3-fold improvement on the sensitivity. Linear ranges of 0.5–50 mg L−1,0.056–5.6 mg L−1 and 0.1–10 mg L−1 are achieved for MTs, Cd-MTs (Cd) and Cd2+, respectively, along with detection limits of 0.013 mg L−1 for Cd-MTs (Cd) and 0.020 mg L−1 for Cd2+. The transformation of cadmium in HepG2 and MCF-7 cells is evaluated after their incubation with Cd2+ reinforced culture medium. Intracellular free Cd2+ cation and Cd-MTs are identified, along with Cd2+ transformation to Cd-glutathione (GSH) adduct/complex, as further demonstrated by ESI-MS.

Using the polymorphism of α-amylase in the culture of winter common wheat, the inheritance of isoenzymes of this enzyme and conjugation of its enzyme types with germinating grain on the “vine,” grain productivity, plant height, and time of ear formation have been studied. It has been shown that the polymorphism of α-amylase isoenzymes in common wheat is limited by the presence of different loci, whose products are similar in electrophoretic parameters. In this regard, one component of the enzyme can be controlled by one, two, or three genes. The locus, which controls α-amylase isoenzyme in the fast moving part of the electrophoretogram, designated as α-Amy-B7 has been identified. The distance of the locus to factor α-Amy-B6 has been determined.

A novel approach to the study of venoms characterizes and compares the venom structure of genus Dendroaspis presented in this work. The complex molecular structure of individual venoms of genus Dendroaspis was defined graphically as a record of synchronous fluorescence fingerprint and atomic force microscopy. Simple comparison of these methods mentioned above of individual analyzed venom samples will immediately reveal changes in each venom composition. Application of these methods with electrophoresis and total protein concentration are new alternatives that were used for monitoring venom composition in selected snakes. Electrophoretogram of black mamba with very low content of total proteins contained the largest number of separated fractions, whereas synchronous fluorescence analysis showed that the highest endogenous fluorescence was found in the venom of black mamba compared with the venom of green mambas, respectively. Our results confirmed different molecular structure in the venom of the genus Den...

The DNA analyst, in order to decide which alleles are present in a sample, uses a graph called the electropherogram (or electrophoretogram), usually referred to simply as the epg The epg represents the separated DNA molecules that are produced after the amplification process of the original sample. It is not necessary here to detail all of the issues affecting the production of the epg as it most rarely materially affects the profile result. However, it is important to understand the general manner in which an epg is produced and how steps can be taken to verify a profiling result. Keywords: epg; electropherogram; separation; alleles; forensic; DNA; profile; graph

African yam bean (Sphenostylis stenocarpa) calcium proteinates and isoelectric proteins were characterized to determine their molecular constitution and functional properties. The electrophoretogram of S. stenocarpa proteins obtained from sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing condition showed that both calcium proteinates and isoelectric proteins had three prominent molecular mass, i.e., 29, 43 and 53 kDa. Calcium precipitation of the protein had a blueshift effect on the protein UV absorption with a shift from 269 nm to 257 nm, whereas UV absorption maximum for isoelectric precipitated protein remained 269 nm as obtainable with flour protein and, thus, no conformational change was associated with isoelectric precipitation. Bulk densities, water and oil absorption capacities of the isolates were in the ranges of 0.76–0.86 g/mL, 2.0–2.33 g/g protein and 0.78–0.99 g/g protein, respectively. Its gelation and water absorption properties were better than or comparable to those of soy bean and some other legumes. PRACTICAL APPLICATIONS African yam bean (Sphenostylis stenocarpa) is a crop grown in some countries in Africa. It produces its edibles (seed grains, root and leaves) in abundance and seems capable of delivering record quantities of protein from soils normally considered marginal. Moreover, the protein is of good nutritional quality. Production of protein isolates from this crop for food formulation and new product development is being envisaged. Its functional attributes examined under this study showed that it might be put to similar use in the food industry as obtainable with soy bean, where high gelling property is of interest in the formulation of value-added food products.

Objectives We compared automated capillary electrophoresis (CE) systems Capillarys 2® from Sebia and V8® from Helena Biosciences Europe (Elitech) with the semi-automated Hydrasys-Hyrys® agarose gel electrophoresis (AGE) from Sebia. Design and methods We evaluated analytical performances and compared 129 fresh routine sera (group A) to 164 frozen pathologic samples with suspicion or antecedent of monoclonal component (MC) (group B). Immunofixation was then compared with immunotyping provided by both CE systems. Results Analytical performances from both CE systems have proven suitable results for clinical use, with within-run and between-run coefficients of variation inferior or equal to 5.2% and 7.7%, respectively. A good correlation was found between AGE and CE with r-value ranging from 0.81 to 0.96 for both CE systems. We observed high MC detection sensitivities (> 85%) of in electrophoretogram readings for both CE systems. MC identification using CE systems provided suitable concordance with immunofixation, although failing to detect some IgM proteins or free light chains. Conclusions Both Capillarys 2® and V8® are reliable automated CE systems for patient care.

The objective of this study was to investigate effect of the steamed soybeans inoculated with Rhizopus oligosporus in advance then Bacillus subtilis on the protein of fermentd soybeans. Soluble nitrogen content and degree of hydrolysis of soy protein increased with increasing fermenting time both of the two-step fermentation and single starter culture of B. subtilis. The amino acid composition of the two-step fermented product fluctuant during fermentation, of which glutamic and aspartic acids had the highest content. However, The most bands of soy protein components with molecular weight above 20 kDa on electrophoretogram of the product by the two-step fermentation became faint. It meant the concentration of these proteins decreased after 24 h of fermentation.

This study investigated purification and milk-clotting activity of the enzymes produced by Bacillus subtilis var, natto and Rhizopus oligosporus compared with that of commercial rennet. The clotting time, viscosity, tension and microstructure of the curd and electrophoretic patterns of milk proteins were determined. The milk-clotting activity/proteolytic activity ratios (MCA/PA ratio) of B. subtilis, R. oligosporus and commercial rennet were also compared. The results revealed that the curd formed by the commercial rennet had the highest viscosity and curd tension and the shortest clotting time among the three enzymes. However, curd produced by Rhizopus enzymes was ranked as second. From the MCA/PA ratio and electrophoretogram analyses it could be concluded that the enzyme produced by B. subtilis had the highest proteolytic activity, while the commercial rennet had the highest milk-clotting activity. Observations of microstructures of SEM showed that the three-dimensional network for curd formed by commercial rennet was denser, firmer and more smooth. The milk-clotting activity, specific activity, purification ratio and recovery of the purified enzymes produced by both the tested organisms were also determined with ion exchange chromatography and gel filtration.

The key steps of the method proposed for isolating the C-terminal fragment of proteins from their tryptic hydrolysates are the esterification of the digest with methanolic HC1 and redigestion with trypsin. The C--terminal peptide is identified by preparing a peptide map using pH 2.1 buffer in the first, and pH 5.5 buffer in the second dimension. The C-terminal fragment appears as an on-diagonal spot. It can be isolated by a pH 5.5 run of the corresponding zone from the first (pH 2.1) electrophoretogram. Yields are up to 45% from different test proteins of known sequences.

Using HCl to activate the diapause eggs is a traditional technique of artificial hatching applied in silkworm egg production. Its mechanism has not yet been clarified. This experiment explored the effect of HCl on the termination of diapause of silkworm eggs cold-stored for 45 days from the point of proteomes. Two-dimensional electrophoresis techniques and ESIMS-MS were used to compare and analyze the variation expression of difficult-dissolved-proteins of diapause eggs coldstored for 45 days before and after acid treatment. Through analysis on the two-dimensional electrophoretogram, there were 296 dots before acid treatment and 302 after the treatment, respectively. Amongst them 265 dots were matchable. The matchability reached 88.6%. There were 31 specific protein dots before acid treatment and 37 after acid treatment, respectively. ESI-MS-MS analysis was conducted for two specific protein-rich dots which disappeared after acid treatment. The results indicated that the sequence of No. 1 protein dot had 55 amino acids' peptide matched with those of chorion protein (Bombyx mori). While the sequence of No. 2 protein dot had only 15 amino acids' peptide matched with those of heat shock protein hsp 19.9 (Bombyx mori), and it was presumed to be an unknown protein. The difficult-dissolvedproteins of diapause eggs have variation expression after acid treatment. Some proteins before and after acid treatment are changed in MW.

In this study, 73 cows with different types of traumatic reticuloperitonitis (TRP) and sequellae along with 25 healthy buffalo cows were used to evaluate the total blood serum protein and protein electrophoresis. According to rumenotomy or necropsy findings, the affected buffalo cows were classified into acute local TRP (AL-TRP), chronic local TRP (CL-TRP), reticular abscess (RA), acute diffuse TRP (AD-TRP), purulent pericarditis (PPC) and fibrinous pericarditis (FPC). Polyclonal gammopathy was characteristic for the electrophoretogram of buffalo cows with CL-TRP (55%), RA (61.1%) and PPC (62.5%). Various degrees of hypoalbuminaemia, hyper-alpha-globulinaemia and low A/G ratios were the associated changes in all cases of TRP and sequellae. Hyper-beta-globulinaemia was noticed in cases with CL-TRP, RA and PPC. Hyper-gamma-globulinaemia was evident in cases with AL-TRP, CL-TRP, RA and PPC. Hyperproteinaemia was noticed in buffalo cows with local complications of TRP including AL-TRP, CL-TRP and RA. Hypoproteinaemia associated with severe hypoalbuminaemia (50% reduction compared to controls) and very low A/G ratios (0.4) characterized buffalo cows with AD-TRP, PPC or FPC. In conclusion, the concentrations and electrophoretic patterns of serum proteins in buffaloes differ according to the anatomical location of the foreign body and the associated pathological lesions.

The physicochemical properties of muscle proteins extracted from two different weight groups (500 and 1500 g) of barracuda ( Sphyraena jello ) are discussed. Solubility of sarcoplasmic protein (SPP) from groups I and II fishes were 107 and 167.18 mg/g, respectively. The myofibrillar protein (MFP) concentration was significantly higher in group II. The concentration of reactive sulphydryl group and Ca 2+ ATPase activity of MFP were higher in group II fishes than in group I fishes. Surface hydrophobicity of MFP was noticeably higher for group I fishes compared to the other group. High emulsion activity index (EAI) was noticed for group I fishes. EAI of SPP increased but that of MFP decreased with increase in protein concentration in both the fishes. MFP from group I fishes showed better emulsion stability (ES) and the ES showed concentration-dependent increase in both groups for both SPP and MFP. Foam expansion was higher in group I fishes, but foam volume stability (FVS) of both the groups did not show any significant difference. Viscosity, water holding capacity (WHC) and gel strength (GS) were higher for group I fishes. Electrophoretogram of SPP and MFP of both the groups show difference in band pattern indicating that proteins expressed during various stages of growth vary.

The electrophoretogram of 89 cats, including those infected by feline immunodeficiency virus (FIV+), feline leukaemia virus (FeLV+) and non-infected, showed statistically significant differences in several of the fractions. FIV+ cats had very high protein values (mean, 8.10 g/dl), mostly because of hypergammaglobulinemia (mean, 2.81 g/dl) as compared with non-infected animals and FeLV+. In addition, in these FIV+ animals, the albumin/globulins ratio (A/G) was very low (mean, 0.72). Statistically significant differences in A/G and alpha2-globulin fraction were observed in FeLV+ group (A/G mean, 0.88 +/- 0.08; alpha2-globulin, mean, 0.84 +/- 0.07 g/dl) when compared with non-infected group (A/G mean, 1.06 +/- 0.08; alpha2-globulin mean, 0.68 +/- 0.04 g/dl). The alpha1-globulin fraction was higher in double infected animals (FIV and FeLV positive, F-F) (3.55 g/dl), than in FeLV+ or FIV+ cats (3.10 and 3.07 g/dl respectively), but no statistical conclusions may be drawn from this fact because of the low number of F-F animals. This technique may help to assess the initial clinical status of retrovirus-infected cats, and the clinical course of these chronic diseases, specifically during and after suitable therapy.

Bisalbuminemia is a rarely encountered anomaly characterized by presence of bifid albumin bands or a single widened albumin band in electrophoretogram. Inherited bisalbuminemia is quite rare and inherited as an autosomal dominant form. The acquired form of bisalbuminemia is usually transient and may be observed during long term beta lactam antibiotic therapy, acute pancreatitis, myeloma and nephrotic syndrome. This is a case of bisalbuminemia in 61-year-old diabetic female with hypothyroidism came with acute exacerbation of bronchial asthma.

Hemoglobin A1c (HbA1c) is now considered to be the marker of choice in diagnosis and management of diabetes mellitus, based on the results of certain landmark clinical trials. Herein, we report the case of a 52-year-old ethnic Southeast Asian Indian man with impaired glucose tolerance whose glycated hemoglobin (ie, HbA1c) levels, as measured via Bio-Rad D10 high-performance liquid chromatography (HPLC) and Roche Tina-quant immunoassay were 47.8% and 44.0%, respectively. No variant hemoglobin (Hb) peak was observed via the D10 chromatogram. We assayed the patient specimen on the Sebia MINICAP capillary electrophoresis platform; the HbA1c level was 6.8%, with a large variant Hb peak of 42.0%. This finding suggested the possible presence of the heterozygous Hb Hope, which can result in spuriously elevated HbA1c results on HPLC and turbidimetric immunoassays. Although the capillary electrophoresis system was able to identify the variant, the A1c results should not be considered accurate due to overlapping of the variant and adult Hb peaks on the electrophoretogram reading. Hb Hope is usually clinically silent but can present such analytical challenges. Through this case study, we critically discuss the limitations of various HbA1c assay methods, highlighting the fact that laboratory professionals need to be aware of occurrences of Hb Hope, to help ensure patient safety.

This study was conducted to evaluate the effect of pepsin digestion at different temperatures (4, 12, 18 and 24 °C) and times (24, 36, 48 and 72 h) on properties of telopeptide-poor collagen from bird feet (TPCBF). The yield, SDS-PAGE, in vitro fibrillogenesis and denaturation temperature of the TPCBF from different treatments were determined. The results showed that the TPCBF yield increased as temperature and time increased. The yield of lots treated at 4 °C were significantly lower than those of the treatments at 12, 18 and 24 °C. In the SDS-PAGE electrophoretogram, the low molecular fragments of samples treated at 12, 18 and 24 °C were higher than those at 4 °C. The fibrillogenesis rate of lots at different temperatures decreased as treated time increased. The denaturation temperatures of the samples treated at 24 °C for 48 and 72 h were significantly lower than those of the treatments at 4, 12 and 18 °C, for all times. These changes could be related to telopeptide removal and lost molecular integrity of collagen by pepsin digestion at a high temperature and long time of treatment. However, the optimum conditions of pepsin treatment should be controlled at 12 °C for 24 h due to the best fibril-forming capacity, high denaturation temperature and an acceptable yield shown in this study.

Asymmetric transformation reaction of L-histidine to D-histidine was studied by homemade capillary array electrophoresis for the first time. The enantiomeric excess value of asymmetric histidine products can be directly determined from the electrophoretogram of capillary array electrophoresis. The experiment results showed that the optimized asymmetric transformation reaction condition was in the presence of salicylaldehyde as catalyst and acetic acid as solvent.

One asymmetric transformation reaction of L-proline (L-Pro) to D-proline was studied by a home-made capillary array electrophoresis (CAE) for the first time. The aldehyde catalysts and the organic acid solvents for the asymmetric transformation reaction were rapidly screened and the enantiomeric excess values of the asymmetric product of L-Pro were directly obtained from the electrophoretogram of CAE.

The tissue distribution of products of 35 enzymatic loci (19 enzyme systems and soluble protein) was examined with different buffer systems and gel thicknesses to identify optimal polyacrylamide gel electrophoretic conditions for allozymic and isozymic studies of the black carp (Mylopharyngodon picius), grass carp (Ctenopharyngodon idellus), silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichys nobilis). Relative levels of enzyme activity are reported for extracts prepared from brain, eye, heart, kidney, liver and muscle. The results showed that the difference within species tissues was greater than that among species; the results showed that the differences of activity between tissues within species was greater than among species; however Acp, Adh, Amy, Cat, Est, Ichh, Gtdh, Mdhp, Pox, Iddh and SP activities present differences among the four species and could be used as genetic markers of species. The electrophoretogram characteristics of Cox, Ldh and G6pdh can be employed as the markers for distinguishing the Leuciscinae and Hypophthalmichthyinae. Copyright © 2005 Society of Chemical Industry

One of the important fields in cell biology is physical stimulus on the growth of plant cells and organs. The effects of strong sound wave on thermodynamic properties, cell cycles and calcium ion signaling transduction of plants have been studied. In this work, chrysanthemum seedlings are treated by sound wave with a certain intensity (100 db) and frequency (1000 Hz) for 3, 6, 9, 12 and 15 days, respectively, and each day for 60 min. The research results show that the activities of protective enzymes and the peroxidase isoenzymes increase at varying degrees. Compared with the control group, there is no new enzyme band in POD electrophoretogram for experimental groups. Our experimental data imply that better promotion for chrysanthemum is obtained under sound wave for 9 days.

In pollen culture methods developed for Nicotiana rustica and N. tabacum, it is possible to induce embryogenic dedifferentiation, i.e., transformation process from immature pollen to embryogenic cells. As biochemical markers for the dedifferentiation of pollen attention has been focused on several phosphoproteins which characteristically appeared in the two-dimensional gel electrophoretogram of total protein from pollen fed with 32Pi in both species. Here, the six phosphoproteins of N. rustica, NrEP1 to 6, were partially purified and their N-terminal amino acid sequences were identified. The sequences were similar to each other and also to those of the phosphoproteins (NtEPs) previously identified in N. tabacum. This suggests that a protein family possessing a common structure plays an important role in the induction process of pollen embryogenesis in Nicotiana.

Made available in DSpace on 2015-02-02T12:39:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-06-01Bitstream added on 2015-02-02T13:08:11Z : No. of bitstreams: 1 S0102-09352014000300665.pdf: 578483 bytes, checksum: 47da50c781afc186539c1989ea3230d4 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Na cólica equina, os estágios iniciais da inflamação incluem alterações denominadas resposta de fase aguda. O objetivo deste estudo foi contribuir para o estabelecimento de valores de referência do proteinograma do líquido peritoneal de equinos hígidos e daqueles submetidos à obstrução intestinal experimental. Vinte e um animais foram distribuídos nos grupos: obstrução de duodeno (GD), íleo (GI), cólon dorsal esquerdo (GM) e controle instrumentado (GC). As colheitas das amostras de líquido peritoneal foram realizadas antes (T0), durante as obstruções (T3) e após as desobstruções (T6, T30, T102 e T174 horas). A proteína total foi determinada pelo método do biureto, e as frações proteicas obtidas por eletroforese em SDS-PAGE. Identificaram-se as proteínas de fase aguda (PFA): IgA, ceruloplasmina, transferrina, albumina, α1-antitripsina, cadeias pesada e leve de imunoglobulina-G, haptoglobina, alfa-1-glicoproteína ácida e uma proteína nominalmente não identificada, que foi chamada P24. Não houve diferença (P>0.3) nas concentrações proteicas entre os grupos, somente entre tempos dentro de cada grupo (P>0.05), evidenciando uma resposta maior das PFA dos grupos obstruídos. O fracionamento eletroforético das PFA, presentes no líquido peritoneal, foi padronizado de modo a estabelecer a curva-padrão para equinos hígidos e para aqueles submetidos à obstrução intestinal; ademais, verificou-se que o referido fracionamento proteico mostrou-se sensível e eficaz no auxílio ao diagnóstico de processos inflamatórios abdominais. The initial inflammatory stages of the colic syndrome include changes known as acute phase response. The aim of this study was to contribute with the establishment of reference values concerning the electrophoretogram of peritoneal liquid from healthy horses and horses submitted to experimentally induced intestinal obstruction. Twenty-one horses were allotted in four groups: duodenal obstruction (DG), ileum obstruction (IG), left-dorsal colon obstruction (MG), and control group (CG). Peritoneal liquid was sampled before obtruction (T0), with 3 hours of obstruction (T3) and 6, 30, 102 and 174 hours after desobstructing (T6, T30, T102 and T174, respectively). Total protein levels were determined by the biuret method and protein fractions were obtained by SDS-PAGE electrophoresis. The acute phase proteins (APP) identified were Immunoglobulin-A, ceruloplasmin, transferrin, albumin, α1-antitrypsin, heavy and light chains of immunoglobulin-G, haptoglobin, α1-acid glycoprotein and a still unnamed protein, which was called P24. There was no difference (P>0.3) in protein levels among groups, although a significant difference (P>0.05) was observed between distinct experimental moments in each group evidencing a higher response of the APP in the obstructed groups. The APP fractioning of the peritoneal liquid was standardized to establish a standard curve for healthy equines and those submitted to induced intestinal obstruction. Moreover, it was verified that the SDS-PAGE electrophoresis was sensitive and effective to help diagnose abdominal inflammatory processes. UNESP Faculdade de Ciências Agrárias e Veterinária Universidade Estadual do Norte Fluminense Darcy Ribeiro Centro de Ciências e Tecnologias Agropecuárias UNESP Faculdade de Ciências Agrárias e Veterinária

Mutations of K-ras gene play an important role in neoplastic progression. The capillary electrophoresis-single strand conformation polymorphism (CE-SSCP) technique is available for the detection of gene mutations. Using an automated capillary electrophoresis with short-chain linear polyacrylamide, after denaturation of PCR products, injections were performed at reverse polarity of 5 kV for 15 s and the separations were carried out under a constant voltage of 8 kV. Of 16 specimens of lung cancer tissue, two specimens were found to have abnormal peaks in the electrophoretogram. CE-SSCP is rapid, automated, and has high performance.

Contents The study was designed to evaluate the influence of season on semen characteristics and seminal plasma protein profile of buffalo bull semen. Thirty-six ejaculates were collected in three seasons (winter, summer and rainy) from six adult Bhadawari bulls, and semen characteristics were evaluated immediately after collection. The seminal plasma was harvested by centrifugation and protein profiling, and percentage protein fractions were analysed by SDS-PAGE. The significant effect of season was observed on ejaculate volume, sperm concentration, progressive motility, percentage live spermatozoa, hypo-osmotic swelling test (HOST) and acrosomal integrity. The electrophoretogram of seminal plasma proteins revealed 20 protein bands in winter, 23 bands in rainy and 25 bands in summer seasons, illustrating the significant effect of seasons on seminal plasma proteins. Among these protein bands, 18 bands were observed common in semen samples of all three seasons while protein bands of 46, 55, 58, 144 and 160 kDa were found in rainy and summer seasons. The protein bands of 48 and 60 kDa were observed only in winter season, whereas 184 and 200 kDa were reported in summer season only. The protein fractions (protein%) of common protein bands observed in three seasons revealed a significant effect of season on protein bands of 24.5, 66, 70, 72, 84 and 86 kDa. From the study, it was pertinent that bull seminal plasma contains specific proteins in particular season, which may be associated with some of the semen characteristics, and these proteins could be used as markers of the semen quality of buffalo bulls.

An intraperitoneal administration of PB at a daily dose of 50 mg Kg-1 body wt for 4 days increased the specific content of hepatic microsomal heme, cytochrome P450 and the activity of aminopyrine N-demethylase by 1.8, 2.8 and 3.5 fold respectively. These results were substantiated by the intensification of the 52.5 KDa polypeptide in the electrophoretogram of sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the hepatic microsomes obtained from PB-pretreated versus control macaques. PB did not affect the hepatic content of cytochrome b5 and the activity of NADPH cytochrome c reductase, whereas it decreased the activity of NADH cytochrome c reductase in the rhesus monkeys. To the best of our knowledge this is a first report on the induction of hepatic cytochrome P450 and related enzymes by PB in rhesus monkeys.

Proteins were extracted and separated by SDS-PAGE from leaves of scions and rootstocks of different Vitis cultivars. Banding patterns were used to identify the different cultivars after extraction of the proteins with a medium containing Tris-HCI (0.68 M, pH 7.8), SDS (2%), β-mercaptoethanol (5%), glycerol (10%) and PEG 20 000 (2%). Electrophoretogram formulae using relative band mobility and staining intensity of the different bands were calculated for the different cultivars. No differences in Rm values were found between individual scion or rootstock cultivars but the technique can be used to distinguish between scion and rootstock cultivars.

We previously found that Curcuma plants and drugs derived from Curcuma longa, C. phaeocaulis, C. zedoaria, and C. aromatica could be identified by the nucleotide differences at two sites and the existence of a 4-base indel on trnK gene. In this paper, based on species-specific nucleotide sequences, the application of a new method, single-nucleotide polymorphism (SNP) analysis was investigated to identify Curcuma plants more conveniently. First, three types of reverse primer were synthesized in different lengths, 34 mer, 26 mer, and 30 mer, to anneal the template DNAs from each species at sites immediately upstream from substitution positions 177 and 645, and at the site including the 4-base insertion from 728 to 731, respectively. After single-base extension reaction of these primers using fluorescent-labeled ddNTPs and PCR products of the trnK gene region as template, the resulting products were detected using an ABI PRISM 310 Genetic Analyzer. The electrophoretogram showed three or two peaks at different positions depending on the 27 mer, 31 mer, and 35 mer product lengths. Each peak was derived from the incorporated fluorescent-labeled ddNMPs complementary to template nucleotides at positions 645, 724, and 177, respectively. C. phaeocaulis showed three peaks of ddCMP, ddAMP, and ddAMP. The other three species showed two peaks derived from 27 mer and 35 mer products: peaks of ddCMP and ddAMP in C. longa, those of ddCMP and ddTMP in C. zedoaria, and those of ddTMP and ddAMP in C. aromatica. Thus SNP analysis to identify four Curcuma plants was newly developed.

Esterases in nine rat hepatic and hepatoma-derived cell lines and normal rat liver homogenate were detected by polyacrylamide gel electrophoresis coupled with active staining with a-naphthyl acetate or butyrate as a substrate. The esterase band patterns of the non-cancerous and oncogene-transformed cell lines were alike, but different from those of hepatoma cell lines and normal rat liver homogenate. The former groups of cells might have completely lost the characteristics of rat liver parenchymal cells, or else they might have their origin at cells other than liver parenchyma. The esterase patterns of the hepatoma cell lines (e.g., McA-RH7777) and the normal rat liver highly resembled with each other, exemplifying the slight biochemical deviation of cancer from normal cells. However, two-dimensional electrophoretogram for the McA-RH7777 cell line showed a prominent esterase spot {p/ 6.0-Mr 110 kDa) that was lacking in the normal liver. This result indicates that there is invariably some change in esterase expression between the cancer cells and the normal liver cells

To verify the performance of crop classification and variety clustering based on electrophoretogram and investigate vegetable seed genetic relationships, three kinds of breeder's seeds samples are collected. They are bell pepper, Chinese cabbage and cucumber. 30 varieties of each kind of crop have been collected. And standard electrophoretograms of them are prepared by method of protein ultra thin isoelectric focusing electrophoresis. Then, the digital images of them are obtained by scanners. Using these images, a pattern recognition model of crop recognition based on PCA and SVM are constructed. Test results by leaving-one method show that, more than 97% crops can be recognized correctly. In addition, through k-means clustering analysis, clustering trees of these 3 kinds of crops with different varieties are established and the paternities of partial varieties based on clustering tree are discussed. This study has positive significance to automatic seed inspection by computer based on electrophoresis and selection of breeding direction.