Electrophoretic Patterns Research Articles

Biochemical characterization and technofunctional properties of bioprocessed wheat bran protein isolates

The effect of three combinations of bioprocessing methods by lactic acid fermentation, cell wall hydrolyzing enzymes and phytase on the biochemical (protein, fat, carbohydrate composition) and technofunctional properties (protein solubility, emulsifying and foaming properties) of wheat bran protein isolates were evaluated. The bioprocessing increased the protein (up to 80%) and fat content (up to 22.8%) in the isolates due to the degradation of starch and soluble pentosans. Additional proteins, globulin 3A and 3C, chitinase, β-amylase and LMW glutenins, were identified from the electrophoretic pattern of the protein isolate bioprocessed with added enzymes. Generally, the bioprocessed protein isolate had lower protein solubility and stronger net charge in pH below 7, when compared to the protein isolate made without bioprocessing. The emulsifying properties of the protein isolates were not affected by bioprocessing. However, the foaming stability of the protein isolates was nearly doubled by bioprocessing with cell wall hydrolyzing enzymes and phytase.

Increased level of fibrinogen chains in the proteome of blood platelets in secondary progressive multiple sclerosis patients.

Epidemiological studies indicate a high risk of stroke, heart failure and myocardial infarction in patients with multiple sclerosis, especially in its secondary progressive (SPMS) phase. Some ischaemic events are directly associated with abnormal platelet functions and their prothrombotic activity. Recent reports, including this study, confirm the increased activation of circulating platelets in SPMS, and also show increased platelet reactivity, among other responses, as well as strong aggregation. In this current study, we conducted a comparative analysis of the platelet proteome in SPMS patients and in healthy controls, to demonstrate the quantitative and qualitative differences likely to affect functional changes observed in SPMS. During densitometry evaluation of 2‐D fluorescence difference gel electrophoresis, we observed differences between the electrophoretic patterns of SPMS platelets and the control samples. To determine a detailed characterisation of the proteome changes in the SPMS patients’ blood platelets, in the next stage, we performed mass spectrometry of selected spots and indicated the increased presence of four proteins (fibrinogen, α‐2 macroglobulin, septin‐14 and tubulin β‐1 chain). The most important of these is the increased amount of prothrombotic protein, fibrinogen, which seems to confirm the accuracy of the imaging and potentially explains the increased risk of platelet‐origin thrombotic events. This study provides new knowledge of the potential existence of the molecular mechanisms responsible for the acceleration of the platelet pro‐coagulant function in SPMS. This can help to identify new targets for therapy, which can then be used not only in the second stage of the disease.

Simultaneous responses of photosystem II and soluble proteins of rapeseed to cold acclimation

This experiment was conducted to assess the quantitative and qualitative changes in soluble proteins as well as some chlorophyll fluorescence parameters in the leaves of a winter canola (Brassica napus L., cv. Licord) under continuous low temperature. Over the experiment, seedlings were initially grown at 15/10 °C (d/n). At fourth fully expanded leafy stage (day 30), a part of the plants were transferred to 4/2°C for 4 weeks. Plants were sampled for protein extraction from leaves in which chlorophyll fluorescence parameters (Fo, Fv, Fm, Fv/Fo, Fm/Fo, Fv/Fm, Fo´, FV´, Fm´ and some other calculated) were also measured. The results showed a clear increase in soluble proteins quantity caused by cold treatment. The enhancements appeared abruptly following the cold exposure to 4°C and lasted. The electrophoretic protein patterns showed changes in the intensity of some polypeptides, besides, induction a new probable protein weighing 47-kW in response to cold treatment. Cold-triggered reduction in maximum quantum yield of PSII (Fv/Fm) was connected especially with drastic decreasing Fv and Fm. Interestingly, high quantitative amounts of soluble proteins along with induction of the new probable polypeptide induced at cold temperature, were attributed to low deduction of maximum quantum yield of PSII. Additionally, more imperative chlorophyll fluorescence parameters changed e.g. qP, NPQ, qL, Y(II) or фPSII etc at light. Nowadays, radar charts or spider plots are the most sophisticated multivariate statistical tools representing physiological responses of plants to abiotic stress conditions or even morphophysiological studies of plants. In rapeseed many researches performed by applying the radar charts for low temperature stresses and interpreted their effects more advancely than common statistical tools. We observed a good representation of the chl fluorescence parameters fluctuations using radar plots. Overall, cold-induced soluble proteins accumulated after longer cold-acclimation, can contribute in photosynthetic apparatus protection against low-temperature damages.

Characterization and antifungal activity of chitosanase produced by Pedobacter sp. PR-M6

To investigate the temperature requirements of chitosanase activity, as well as the degradation patterns generated by enzyme-induced chitosan oligomer hydrolysis, Pedobacter sp. PR-M6 was inoculated onto 0.5% colloidal chitosan medium agar plates. Cell growth was higher at 30 °C than at 20 °C during the initial 2 days of incubation. The protein content rapidly increased on day 1 at both temperatures and then it slowly increased at 20 °C and slowly decreased at 30 °C during the following 5 days of incubation. In order to characterize the electrophoretic pattern, Pedobacter sp. PR-M6 was cultured in 1% powder chitosan medium at 20 °C and 30 °C for 5 days after incubation and analyzed by SDS-PAGE. Four bands were visible, corresponding to ct1 (25 kDa), ct2 (17 kDa), ct3 (15 kDa), and ct4 (14 kDa), at both 20 °C and 30 °C. The optimal conditions for the activity of chitosanase produced from Pedobacter sp. PR-M6 were 60 °C and 1.81 enzyme units/mg protein. Two major isozyme bands (ct3 and ct4) exhibited their strongest chitosanase activity at 50 °C in SDS-PAGE gel. The reaction products generated from (GlcN)2-(GlcN)5 substrates at 60 °C after a 1 h incubation were investigated by thin-layer chromatography. Low-molecular weight chitosan and oligochitosan (LCOC) and soluble chitosan showed antifungal activity against A. brassicicola, B. cinerea, F. solani, and R. solani. LCOC exhibited higher antifungal activity than soluble chitosan. Moreover, LCOC treatments (500 ppm and 1000 ppm) inhibited conidia germination in A. brassicicola.

Optimising detection of acidification kinetics diversity in Lactococcus lactis subsp. lactis using SDS‐PAGE protein fingerprinting as screening method

Cell‐free extracts from 157 Lactococcus lactis strains isolated from Artisan cheese were screened by protein fingerprinting previous to their technological characterisation. The strains were classified according to their electrophoretic patterns into five groups. A set of strains representing the different clusters were selected to study their acidifying activity in milk. Time and rate feature points, as well as the shape of the acidification curves, resulted in six different fermentation kinetics, mostly consistent with the electrophoretic groups. Thus, selection of native strains as starter cultures based on their acidifying activity could be optimised by protein fingerprinting.

Quantification of Micrornas by Absolute Dpcr for the Diagnostic Screening of Colon Cancer

There is currently no validated micro(mi)RNA diagnostic stool test to screen for colon cancer (CC) on the market because of the complexity of fecal density, vulnerability of stool to daily changes, and the presence of three sources of miRNAs in stool (cell-free from fecal homogenates, exsosomal miRNAs from fecal exosomes, and fecal colonocytes). To address these complexities, we have first carried out a microarray miRNA experiment, using Affymetrix GeneChip miRNA 2.0 Arrays, on immunocaptured and enriched stool colonocytes of 15 subjects (three healthy controls and twelve colon cancer patients [three TNM stage 0-1 (e.g., polyps ³ 1 cm, villous or tubvillous, or with high grade dysplasia), three stage 2, three stage 3, and three stage 4 in triplicates to select a smaller panel of 14 preferentially expressed mature miRNAs associated with colon cancer (12 Up-Regulated, miR-19a, miR-20a, miR-21, miR-31, miR-34a, miR-96, miR-106a, miR-133a, miR-135b, miR-206, miR-224 and miR-302; and 2 Down-Regulated, miR-143 and miR-145). In a subsequent validation study carried out on total small RNA extracted by immunocapture, followed by RT that employed TaqMan® miRNA Reverse Transcription (RT) Kit and a Custom TaqMan RT Primer Pool, absolute quantification of miRNAs, in copies/µl, was measured using a chip-based Absolute QuantStudio 3D Digital PCR analysis. To ensure that we have chosen human and not bacterial small total RNA, we have carried out coextraction protocols with E. coli K1 strain RS18, compare Agilent electrophoretic patterns, and also sequenced random samples throughout this research using mRNA/miRNA sequencing. Our initial quantitative dPCR miRNA data presented herein showe that the quantitative changes in the expression of a few mature miRNA genes in stool, which are associated with right and left colon cancer, would provide for a more convenient, sensitive and specific diagnostic screening markers thatare more useful than those test markers currently available on the market, such as the low-sensitivity (<15%) fecal occult blood test (FOBT); result in better compliance; and is more economical than the invasive and expensive colonoscopy exam in colon cancer, which can be cured if that cancer is detected at the early TNM stages, and that becomes incurable and deadly if not diagnosed before metastasis. Initial test performance characteristics of the miRNA approach showed that the test has a high numerical predictive value in colon cancer. Moreover, underpinning of the miRNA markers as a function of total RNA showed that the test can numerically differentiate between control subjects and colon cancer patients, particularly at the early stages of that curable cancer. We propose to extend our initial research results to a larger prospective and randomized five-years nested case-control study, to validate the expression of the above 14 miRNAs, in stool of 180 individuals in an epidemiologically designed study, using (30 controls and 150 colon cancer patients (thirty precancerous polyps (stage 0-1), forty five stage 2, and seventy-five colon cancer stages 3 or 4). chosen randomly by an epidemiological method from 900 control and CC subjects to allow for an adequate time to collect the required 900 stool samples, as well as allowing for statistically valid analysis, standardized test conditions, and to provide a mean for determining the true sensitivity and specificity of a miRNA-screening approach in noninvasive human stool. Power-analysis has indicated that a total of 180 individuals, which will take us 5 years to enroll in testing, is an appropriate number of subjects to standardize and validate our proposed miRNA screening test. We may find out at the end of the proposed validation study in stool that fewer miRNAs, or even one miRNA, may suffice to serve as an efficient and a quantitative marker for the non-invasive diagnostic screening of colon cancer in human stool. The above approach when combined with bioinformatics analysis, to correlate miRNA seed data with our previously published messenger (m)RNA target data in stool, allows for a thorough mechanistic understanding of how miRNA genes regulate mRNA expression, and would offer a better comprehensive diagnostic screening test for the non-invasive early detection stage (0-1) of colon cancer. In order to show the clinical sensitivity and specificity of the proposed miRNA test, the absolute miRNA PCR values, in copies/µl, will be correlated with FOBT, colonoscopy, and pathology data. Standardization will establish test’s performance characteristics (sample selection, optimal sample running conditions, preservation and storage) to ensure that the assay will perform the same way in any laboratory, by any trained personnel, anywhere in the World. Ultimately, a smaller number of selected validated miRNAs (<10) showing increased and reduced expression could suffice to give quantitative miRNAs colon cancer expression values, useful for the early diagnostic screening of that curable cancer.

Electrophoretic patterns of proteinuria in feline spontaneous chronic kidney disease.

The purpose of this study was to describe the electrophoretic patterns of proteinuria in cats at risk of and cats with chronic kidney disease (CKD), and to investigate whether the presence of high-molecular-weight (HMW) and low-molecular-weight (LMW) proteins were associated with CKD, proteinuria and/or disease progression. Healthy cats at risk of developing renal disease (n = 17) and cats affected with CKD at different stages (n = 22) were prospectively enrolled and sampled over time. Seventy urine samples were included and assayed with a commercially available sodium dodecyl sulfate-agarose gel electrophoresis (SDS-AGE) method. Each sample (gel lane) was inspected to identify albumin, HMW and LMW proteins, and an electrophoretic pattern (albuminuria, glomerular, tubular, mixed or negative) was assigned accordingly. Fisher's exact test was used to assess the distribution of HMW and LMW proteins in cats grouped according to International Renal Interest Society stage and to the magnitude of proteinuria, and to assess if HMW and LMW proteins at the time of inclusion were associated with the development and progression of CKD. In samples of cats at risk, the most common pattern was glomerular (84.6%); glomerular pattern was also common in cats with CKD (54.2%), although mixed proteinuria and tubular proteinuria were also present (29.5% and 11.4%, respectively). The presence of LMW proteins was associated with CKD (P <0.0001) and to a urine protein:creatinine ratio >0.2 (P = 0.025). Both HMW and LMW proteins were not associated with progression of CKD within 6 months (n = 14). Our results showed that HMW proteinuria is common in healthy cats at risk of developing CKD, although the pathological significance needs to be confirmed. The detection of LMW proteins in urine of cats suspected to be affected by CKD, especially in non-azotaemic, non-proteinuric or borderline proteinuric cats, suggests the presence of kidney damage.

Serum CA 19.9 Levels in Patients With Benign and Malignant Disease: Correlation With the Serum Protein Electrophoretic Pattern.

To confirm that the carbohydrate antigen 19.9 (CA 19.9) protein can be evaluated by determining changes in the β2 zone in protein electrophoresis. A total of 75 patients (64 with cancer, 11 with benign diseases) with abnormal serum CA 19.9 level were included. Patients with cancer had significantly higher serum CA 19.9 concentrations than those with benign diseases (p<0.001). Similar CA 19.9 levels were observed in patients with normal (median 1129 U/ml), weakly positive (median 699 U/ml) and positive (2333 U/ml) β2 fraction in protein electrophoresis. In contrast, changes in the protein pattern of the β2 fraction were related to an inflammatory pattern with significantly higher C-reactive protein concentration (p<0.0001), independently of serum CA 19.9 level. The intensity of the β2 fraction in protein electrophoresis is related to inflammation and not to CA 19.9 in patients with cancer or other diseases.

COLLAGEN EXTRACTION FROM YELLOWFIN TUNA (Thunnus albacares) SKIN AND ITS ANTIOXIDANT ACTIVITY

Tuna skin, a byproduct of the fish processing industry, is used as an alternative collagen source to replace bovine and porcine products. This study aimed to extract collagen from tuna skin with acetic acid, and investigated the antioxidant activity. Collagen extraction was carried out through a pretreatment process, defatted with butyl alcohol, and soaking in acetic acid to extract the Acid Soluble Collagen (ASC). The effect of concentration of sodium hydroxide and soaking time on the non-collagenous protein removed were measured, and evaluated. The yield and antioxidant activity of each sample were evaluated and the best result was determined by ANOVA. The highest yield of collagen was 3.18% based on dry weight reached at the treatment with sodium hydroxide 0.2 M and acetic acid 1 M. The different treatments did not result in any significant differences in the spectrum of amide A, B, I, II and III which are the characteristics spectra of collagen. Based on the electrophoretic pattern, tuna skin collagen has two  chains (1 and 2), and one β chain. Therefore, it is classified as type I collagen. The main amino acids were glycine and proline. In addition, the strongest antioxidant activity was found in the sample treated with sodium hydroxide 0.05 M and acetic acid 1 M treatment with IC50 value of 0.45 mg/mL. This study is the first to report on antioxidant activity from fish collagen (not hydrolysate or peptide products).

Effect of high-intensity ultrasound on the compositional, physicochemical, biochemical, functional and structural properties of canola (Brassica napus L.) protein isolate.

The objective of this study was to investigate the impacts of high-intensity ultrasound treatments on the compositional, physicochemical, biochemical, functional and structural properties of canola protein isolates (CPI). Aqueous canola protein suspensions were sonicated at 40 kHz for 15 min and 30 min. The moisture content, water activity, bulk density and the L* and a* color parameters of the CPI decreased due to the ultrasound; however, the in vitro protein digestibility was not modified by the treatment. Glutelin (57.18%) was the main protein fraction in the canola protein isolate. SDS-PAGE demonstrated that there were no changes in the protein electrophoretic patterns, thus indicating that sonication did not break the covalent bonds. However, the ultrasound treatment improved the protein solubility, oil absorption capacity and the emulsifying, gelation and foaming properties, but these improvements depended on the pH and ultrasound exposure time. Scanning electron microscopy revealed that the ultrasound treatment disrupted the microstructure of the CPI by exhibiting larger aggregates as a lyophilized powder. In addition, there was an increase in the surface hydrophobicity and a decrease in the size of the particles of the canola protein due to the ultrasound effects, which indicates a destruction of the particles or a dissociation of the protein aggregates in the canola protein dispersions. These results suggest that ultrasound treatment is a valuable tool for improving the characteristics of canola proteins for use in foods.

Antioxidant activity of Cheddar cheese during its ripening time and after simulated gastrointestinal digestion as affected by probiotic bacteria

ABSTRACT The purpose of our study was to evaluate the effect of the addition of Lactobacillus casei and Lactobacillus plantarum to Cheddar cheese on the antioxidant properties during its ripening time and after simulated gastrointestinal conditions. During the whole ripening time, the scavenging of DPPH and hydroxyl radicals both reached their maximum activity at the 16th week, while reducing power activity was reached at the 20th week. The antioxidant values of probiotics cheese were significantly higher than that of control (p< 0.05). Similar results were found after simulated gastrointestinal conditions. The simulated gastric digestion can significantly reduce the activity of DPPH radicals-scavenging (p< 0.05), but after intestinal condition, the values came back to the same level compared with that before digestion. The electrophoretic patterns indicated that adding probiotic to cheese could promote the hydrolysis of protein to enhance the antioxidant activity, especially with mixed probiotics. The addition of probiotic bacteria to Cheddar cheese had nearly no effect on its composition, texture profile analysis, and sensory analysis.

Evaluation of an active yeast propagation system on fermentation traits and quality of C.V. Carmenère wine

Active dry yeasts (ADY, Saccharomyces cerevisiae) are widely used in oenology due to their potential benefits on the control of fermentation and quality reproducibility among other aspects. On the other hand, yeast propagation systems, so called Active Yeast Systems (AYS), can be useful to optimize the alcoholic fermentation (AF) initial lag phase and reduce production costs. The objective of this work was to determine the predominance of an ADY strain propagated by AYS and the impact of this inoculum on cv. Carmenère wine quality. Lalvin ICV D21 ADY strain was inoculate according to the protocol recommended by the manufacturer (T0), and in parallel, it was propagate by the AYS and then used as inoculum (T1). Yeast strain predominance analyzed by restriction fragment length polymorphism (RFLP) technique indicates that nine (out of 9) yeast colonies obtained from a single sample of the ADY, show the same electrophoretic pattern when compared to the ADY. The results show limited significant differences for the fermentation speed and the yeast cell counting. The result of the physicochemical analysis of the musts and resulting wines showed no significant differences between treatments. A triangular test showed no significant sensory differences between wines

The electrophoretic pattern of serum proteins in dogs with babesiosis

This study was aimed at the evaluation of the electrophoretic pattern of serum proteins in dogs naturally infected withBabesia canis. Blood samples were collected from 37 dogs infected withB. canisand showing clinical signs consistent with the disease. The sick animals were classified as dogs with physiologic and decreased red blood cell (RBC) values. Twenty-five healthy dogs formed the control group. The concentrations of total proteins and protein fractions were measured in blood serum. The values of total proteins, albumin and albumin/globulin (A/G) ratio in dogs with babesiosis were significantly lower than in healthy ones (P&lt; 0.001). In the globulin fractions, significantly higher relative concentrations of α1-, β1- and β2-globulins (P&lt; 0.01), and non-significantly higher values of α2- and γ-globulins were found in dogs with babesiosis with a double α2-zone in six out of 37 animals. Marked differences were observed also between the two groups of sick animals, with significantly lower values of albumin and A/G ratio (P&lt; 0.05), and significantly higher values of α1- and β1-globulins in dogs with decreased RBC (P&lt; 0.05 andP&lt; 0.01, respectively). Presented results indicate marked alterations in the electrophoretic pattern of serum proteins in dogs with babesiosis suggesting its usefulness for the evaluation of pathophysiological changes caused by the disease and for diagnostic of disease severity.

Effect of incorporation of sodium caseinate, whey protein concentrate and transglutaminase on the properties of depigmented pearl millet based gluten free pasta

The current investigation reports the preparation of gluten free pasta from depigmented pearl millet flour blended individually with sodium caseinate (SC) (10 and 20%) and 1:1 combination of sodium caseinate and whey protein concentrate (SCW) (10 and 20%), both treated with and without transglutaminase (TG) enzyme (0.5 and 1.5%). SC addition (at 10 and 20%) maintained the cooking loss (10.38 ± 0.25 and 11.08 ± 1.71) and yellowness (b*) (16.11 ± 0.29 and 15.79 ± 0.32) as like control sample, decreased the water absorption (1.73 ± 0.19 and 1.47 ± 0.04), brightness (L*) (42.10 ± 0.49) (at 20%), pasting properties, and increased the redness (a*) (2.81 ± 0.10 and 3.58 ± 0.16) and sensory attributes (except colour and adhesiveness). The combination of SCW (at 10 and 20%) decreased the cooking loss (6.90 ± 0.00 and 5.99 ± 0.11), water absorption (1.92 ± 0.05 and 1.72 ± 0.08), pasting properties and increased L*, a*, b* (46.83 ± 0.45, 2.53 ± 0.09, 17.08 ± 0.20) (at 10%) and sensory attributes (except colour and adhesiveness). TG addition increased the cooking loss, water absoprtion and pasting properties (in all the samples). Protein solubility (%) decreased with increase in the enzyme concentration confirmed with electrophoretic patterns. Moreover, the TG treatment imparts tenderness and chewiness to pasta. Hence, the present combination could be useful for preparation of good quality of gluten free pasta.

Physicochemical characterization of a zein prepared using a novel aqueous extraction technology and tensile properties of the zein film

Recently, our research team developed a novel aqueous extraction technology (AET) for preparing an α-zein enriched product from corn gluten meal without any organic solvents. This technology is conspicuously different from the conventional organic solvent extraction technology (OSET). The present study aimed to investigate the physicochemical and film-forming properties of the AET zein in comparison with those of the commercial zein by alcohol extraction. The results revealed that similar to the OSET zein, the AET zein was rich in proteins, especially α-zein. Meanwhile, electrophoretic pattern and amino acid composition were almost the same for both zeins, except that AET zein had trace quantities of β-zein and protein hydrolysates, along with a higher proportion of Ile and lower proportion of His. The intrinsic differences probably contributed to a convergent pattern of the protein solubility in aqueous ethanol between AET zein and OSET zein, and a pronounced higher tensile strength/elongation of AET zein film. In addition, the removal effect of AET on some hazardous heavy metals brought in by raw materials, and no exogenous alcohols in the AET zein were confirmed by atomic absorption spectrophotometry and gas chromatography, respectively. The present findings suggest that AET zein has essential characteristics for use as an potential food ingredient or a pharmaceutical excipient, and the aqueous extraction technology thereby could be considered as an alternative strategy for zein production.

ИСПОЛЬЗОВАНИЕ ЭЛЕКТРОФОРЕЗА БЕЛКОВ ЗЕРНА ДЛЯ КОНТРОЛЯ ГЕНЕТИЧЕСКОЙ ЦЕЛОСТНОСТИ ОБРАЗЦОВ ЯРОВОЙ МЯГКОЙ ПШЕНИЦЫ ИЗ КОЛЛЕКЦИИ ГЕНЕТИЧЕСКИХ РЕСУРСОВ РАСТЕНИЙ ВИР

Background. VIR's Department of Wheat holds the collections of viable accessions and its originals. The process of reproduction and restoration of germination ability in wheat collections requires monitoring of varietal purity and integrity. For this purpose, botanical varieties that describe sets of morphological characteristics of the spike and grain are used. However, sometimes it is necessary to check the conformity of a variety with its original accession. In this case, the gliadin electrophoretic banding patterns are reliable markers in identifying cultivars and biotypes of wheat. Materials and methods . The study included 7 accessions of spring bread wheat: 3 for the cultivar 'Rubin' and 4 for the cultivar 'Novinka'. Identification of accessions by electrophoresis of gliadin storage protein was carried out according to the generally accepted method. Results and conclusions . Comparison of gliadin electrophoretic banding patterns of the varieties from the VIR collection of plant genetic resources with analogous accessions from other genebanks of the world resulted in the recovery of the lost varieties 'Rubin' (Sweden) and 'Novinka' (Russia).

ПЕРСПЕКТИВЫ ИСПОЛЬЗОВАНИЯ ЗЕИНОВЫХ МАРКЕРОВ В СЕЛЕКЦИИ ЛИНИЙ И СОРТОВ ВОСКОВИДНОГО ПОДВИДА КУКУРУЗЫ

Background. Among the numerous maize accessions in the VIR collection, Plant special attention is drawn to the lines and varieties of waxy maize due to their specific feature, that is, the content of high-quality grain starch that consists entirely of amylopectin. This amylopectin starch is of special interest to the medical, starch and syrup, and confectionary industries. One of the most important applications of amylopectin starch in medicine is its use as a component of Volekam blood plasma substitute. Besides amylopectin, the grain of other maize subspecies contains much amylose starch which makes production of purified amylopectin expensive. Therefore, the creation and study of domestic waxy maize lines and varieties is very topical, however it is insufficiently developed at present. Materials and methods . For the first time, zein electrophoresis was used to analyze waxy maize old landraces and breeding lines. Zein electrophoresis was carried out in vertical 10% PAA gels with 8 M urea. The solution for zein extraction contained 6 M urea and 0.01 M dithiothreitol. Results. The present research pursued the following aims: to use zein elecrophoretic patterns (protein markers) in screening waxy maize landraces and lines for their identification and fingerprinting; to determine the biotype composition of landraces and their polymorphism and degree of relationship; and to demonstrate the possibility of applying zein markers in the development of new waxy maize inbred lines by backcrossing the wx1 gene into the initial dent maize line. Conclusions. Zein electrophoretic patterns showed the uniqueness of each waxy maize accession. All the studied new lines are united by a decreased intensity of most zein pattern components, as well as by the presence of an intensive component 47 which is absent in the pattern of the initial lines of dent maize. Component 47 may serve as a protein marker for identifying waxy maize lines. All landraces were found to be composed of several biotypes (from two to five). Regardless of the intravarietal polymorphism, every variety has specific patterns which facilitate its identification.

Influence of multigrain premix on nutritional, in-vitro and in-vivo protein digestibility of multigrain biscuit.

Effect of addition of multigrain premix (MGP) prepared using a combination of cereals, pulses and oilseeds at 40% level, on nutritional properties of multigrain biscuit, its in-vitro and in-vivo protein digestibility and protein profiling were studied. The incorporation of MGP significantly increased the protein content (from 7.37 to 16.61%), insoluble dietary fiber (from 1.71 to 6.67%), soluble dietary fiber (from 0.46 to 2.42%). The significant increase in the levels of isoleucine (ND-34.79%), methionine (0.04 to 7.65%), tryptophan (0.22 to 5.95%) valine (0.38 to 16.58%), lysine (0.36 to 7.32%), and threonine (0.51 to 7.2%) was observed, whereas fatty acid profile of MGP incorporated biscuits showed increased polyunsaturated fatty acids and decreased saturated fatty acids. The vitamin-mineral profile of MGP incorporated biscuits showed increased the thiamin (0.07-0.21mg/100g), riboflavin (0.09-0.28mg/100g), calcium (12.89-45.28mg/100g) and iron (1.13-3.47mg/100g) contents. The in-vitro protein digesibility of multigrain and control biscuits indicated that the proteins present in multigrain biscuits had high digestibility (71.73%) as compared to control biscuit (38.13%). The in-vivo studies indicated that, the protein quality of multigrain biscuits was comparable with casein protein with high protein efficiency ratio of 3.02. The electrophoretic pattern of multigrain biscuits showed subunit molecular weight distribution of different protein units and aggregation of protein bands at high molecular weight region of 85 to 166kD. The outcome of the study indicated the possibility of utilising MGP to improve the overall nutritional quality of biscuits.

Enrichment of common carp (Cyprinus carpio) fingerlings diet with Psidium guajava: The effects on cutaneous mucosal and serum immune parameters and immune related genes expression

The present study aimed at evaluation of possible effects of Psidium guajava as immunostimulant on mucosal and serum immunity, as well as related genes expression in common carp (Cyprinus carpio) fingerlings. In an eight weeks feeding trial, fish were fed with experimental diets containing guava leaf powder [GLP] (0, 0.25, 0.5 and 1%). Then, skin mucus and serum immune parameters (total Ig, alkaline phosphatase activity, lysozyme activity), as well as immune related genes expression (TNF-alpha, IL1b, IL8) were measured. All levels of GLP inclusion in the diet brought about the increase in total Ig in the serum, while in the mucus only the highest level of GLP was effective. No significant differences were observed in mucus alkanine phosphatase activity, while fish fed 0.25% of GLP had noticeably higher serum lysozyme activity. The mucus total protein electrophoretic pattern revealed some differences between the control and the treated fish. Two high MW bands were present in the control group but not in the treated groups. Three bands with a low MW comprised between 35 and 17 mw were noticed in GLP fed carps and not in the control. Gene expression studies revealed no noticeable alterations in TNF-alpha gene between treated fish and control. However, the level of expression of IL-8 was strongly upregulated in fish fed 0.5 and 1% of GLP. Also, feeding on 0.5% of GLP significantly upregulated IL1b gene expression. Evaluation of growth performance revealed improved performance parameters in GLP treated groups. These results confirmed possible immunomodulatory and beneficial effects of GLP on common carp fingerlings.

Distinction of apple cultivars according to their mal D 1 allergen coding genes using PCR technique

Apple is one of the most important fruit grown and consumed in the temperate climate region. About 2% of the European population suffers from several allergenic reactions after consumption. Presence of 7 members of Mal d 1 gene family occurring in some apples was examined by PCR. The Mal d 1.01 and 1.02 genes could be detected from 91% and 79% of apple cultivars, respectively, due to the high degree of conservative regions. The Mal d 1.04 gene has 4 functional varieties and 2 pseudo-alleles, so it is highly variable. The PCR amplifi cation with Mal d 1.06 primers gave one or two fragments with different sizes. The electrophoretic pattern is a suitable means to select apple cultivars according to their low, medium, or high Mal d 1 allergen content. Florina apple showed the single 154 bp allele, which is responsible for the small Mal d 1 allergen content in homozygote form.. Several samples gave weak signal or did not give any fragment-band on the gel, so Mal d 1.07 and 1.08 genes might have more varieties. Regarding the Mal d 1.09 gene we have found that it has conservative sequences in different apple cultivars and does not have too many varieties.