Electrophoretic Patterns Research Articles

Mucopolysaccharidoses diagnosis in the era of enzyme replacement therapy in Egypt

BackgroundUndegraded glycosaminoglycans (GAGs) induced by deficiency of enzymes are the primary cause of mucopolyscchardoses. Mucopolysacchardoses (MPS) are a group of rare lysosomal storage diseases (LSD). The quantification of a specific enzymatic activity is needed for accurate diagnosis. The objectives of this work were: first, to continue the study of mucopolysacchardoses disease in Egypt after the start of using the enzyme replacement therapy (ERT). Second, to define the commonest types among our population after 18 years experience with the disease. Third, to compare the different MPS types’ distribution, diagnosed after the start of the ERT, to identify the impact of using ERT on the number and type of diagnosed patients. MethodUrinary GAGs were measured for all referred cases followed by two-dimensional electrophoretic separation for cases with high levels of GAGs; the specific enzyme activity was assayed for each type depending on the abnormal electrophoretic pattern obtained. Clinically suspected cases of Morquio syndrome were directly subjected to measuring the specific enzyme. ResultsOut of 1448 suspected cases, 622 (42.9%) MPS patients were diagnosed revealing the following distribution: MPS I (172, 27.7%), MPS II (57, 9.1%), MPS III [(177, 28.5%: 134 type B and 43 types A, C or D)], MPS IVA (124, 19.9%), MPS VI (90, 14.5%) and MPS VII (2, 0.3%). MPS III was the most commonly diagnosed type followed by MPS I and MPS IVA. MPS IVA represented the most common type receiving treatment, followed by MPS I, MPS II and MPS VI. ConclusionThe presence of treatment encouraged the affected families and physicians to seek diagnosis. MPS III was the commonest type among our studied group after 7 years of diagnosis, while MPS IVA was the commonest type receiving treatment.

Potential application of DBD Plasma Technique for modifying structural and physicochemical properties of Soy Protein Isolate

The present study was designed and implemented to modify soy protein isolate (SPI) using dielectric barrier discharge (DBD) plasma and investigate the effects of this treatment on the protein's structural and physicochemical properties. For this purpose, SPI powder was treated with DBD plasma for 0, 5, 10, 15 min at 16, 18, 20 kV. Then, solubility, water holding capacity (WHC), emulsifying activity and emulsion stability, foaming capacity and foam stability, total and reactive sulfhydryl, free carbonyl, FTIR, electrophoretic patterns, as well as protein structures were evaluated. Results indicated that emulsifying properties, solubility, WHC, and foaming activity of SPI were enhanced, and the highest values were obtained after plasma treatment at 18 kV for 15 min. Free and reactive sulfhydryl and free carbonyl contents increased under prolonged exposure to DBD plasma at the applied voltage of 18 kV (p < 0.05). FTIR and circular dichroism (CD) results indicated partial denaturation and unfolding of the α-helix structure of SPI followed by the formation of protein aggregates resulting from 15 min plasma exposure; microscopic images also confirmed these results. The above changes indicate successful modification of the physicochemical and functional properties of SPI treated with DBD plasma.

The Proteolytic Inactivation of Protein Z-Dependent Protease Inhibitor by Neutrophil Elastase Might Promote the Procoagulant Activity of Neutrophil Extracellular Traps in Sepsis.

Septic shock is the archetypal clinical setting in which extensive crosstalk between inflammation and coagulation dysregulates the latter. The main anticoagulant systems are systematically impaired, depleted, and/or downregulated. Protein Z-dependent protease inhibitor (ZPI) is an anticoagulant serpin that not only targets coagulation factors Xa and XIa but also acts as an acute phase reactant whose plasma concentration rises in inflammatory settings. The objective of the present study was to assess the plasma ZPI antigen level in a cohort of patients suffering from septic shock with or without overt-disseminated intravascular coagulation (DIC). The plasma ZPI antigen level was approximately 2.5-fold higher in the patient group (n = 100; 38 with DIC and 62 without) than in healthy controls (n = 31). The elevation's magnitude did not appear to depend on the presence/absence of DIC. Furthermore, Western blots revealed the presence of cleaved ZPI in plasma from patients with severe sepsis, independently of the DIC status. In vitro, ZPI was proteolytically inactivated by purified neutrophil elastase (NE) and by NE on the surface of neutrophil extracellular traps (NETs). The electrophoretic pattern of ZPI after NE-catalyzed proteolysis was very similar to that resulting from the clotting process-suggesting that the cleaved ZPI observed in severe sepsis plasma is devoid of anticoagulant activity. Taken as a whole, our results (1) suggest that NE is involved in ZPI inactivation during sepsis, and (2) reveal a novel putative mechanism for the procoagulant activity of NETs in immunothrombosis.

Assessment of Genetic Diversity in Bambara Groundnut (Vigna subterranea) Landraces Based on Seed Storage Proteins Electrophoretic Pattern

Bambara groundnut (Vigna subterranea) is a leguminous crop that is considered underutilized and has previously received little research attention. Variability in a number of physiological factors, including germination rate, widely affects its production. Seed storage protein, its fractions and protein profile of six Bambara groundnut local landraces were studied to assess their genetic relatedness. Total seed storage protein and its fractions were estimated by Bradford’s method. SDS-PAGE analysis was used to evaluate storage protein profile. The results showed significant differences (p&lt;0.05) in protein contents among the landraces. The major seed storage proteins were found to be globulins (0.048 to 0.088mg/mL ), albumins (0.023 to 0.038mg/mL ), glutelins (0.007 to 0.013mg/mL ) and prolamins (0.002 to 0.004mg/mL ). Five peptide bands were detected with molecular weights corresponding to 97.4 kDa, 45 kDa, 29 kDa, 20.1 kDa and 18 kDa, respectively. Three peptide bands corresponding to 97.4 kDa, 45 kDa and 18 kDa were detected in all the landraces and two peptide bands between 29 kDa and 20.1 kDa were detected in five landraces. Dendrogram generated by UPGMA grouped the six landraces into one major cluster with two sub-clusters. The observed diversity in storage protein pattern of the landraces indicated their potential as materials for crop improvement.

Investigation of the compositional and structural changes in the proteins of cow milk when processed to cheese

Various treatments applied during dairy processing strikingly affect milk proteins, the major structural determinants, thus causing modifications in some techno-functional and biological properties of the final dairy product. Cheeses, with a variety of types, are mostly consumed processed dairy foods worldwide. The purpose of this study was to investigate the effect of heating, culture/enzyme activity and other treatments on the composition and molecular structure of cow milk proteins during the processing of different cheeses through electrophoresis, chromatography and spectroscopy. Electrophoretic patterns of the samples successfully indicated differences in protein content through process steps and in the final products. Curdling after pasteurization at 85 °C revealed many newborn peptides not detected in raw cow milk and other dairy samples. High and low similarities in peptide profiles of cheeses to that of raw cow milk indicated high-temperature pasteurization-induced unfolding facilitated extensive proteolysis subsequently. Significant conformational changes including loss of α-helices and exposure of β-strands observed in the secondary structure of proteins especially through curdling might offer alterations of reactive/functional groups. In total, pasteurization, curdling, salting, and ripening treatments remarkably altered the content and structure of cow milk proteins through unfolding, denaturation and aggregation, strongly suggesting modified functionality when milk was processed to cheese.

Sensitivity of midgut physiological parameters of Lymantria dispar L. larvae to benzo[a]pyrene in populations with different multigeneration contact to environmental pollutants

Accumulation of organic pollutants in the environment calls for sensing physiological parameters adequate to indicate the presence of contaminants and their effects on ecosystems. Evidence points to the importance of insect adaptations in their habitats for the assessment of sensitive biomarkers so we examined the influence of origin and multigenerational adaptations of the Lymantria dispar larvae to chronic benzo[a]pyrene (B[a]P) treatment under laboratory conditions. The main aim was to compare reactions of larvae from unpolluted and polluted forests using alkaline phosphatase (ALP), acid phosphatase (ACP), and carboxylesterase (CE) specific activities in the midgut, including electrophoretic isoform patterns; midgut expression levels of Hsp70, larval development time (DT), and midgut mass (MM), after chronic exposure to 5 and 50 ng of B[a]P/g dry food weight. The biomarker potential of these parameters regarding larval pre-exposure history to pollution was estimated by principal component analysis (PCA). B[a]P treatment resulted in inhibition of ALP activity, a rise of CE activity, and reduction of MM in larvae from the unpolluted forest, while the population from the polluted forest showed significant elevation of Hsp70 expression in the midgut, prolonged DT, and reduction of MM. PCA confirmed variations in responses of the selected parameters regarding population origin. The obtained results provide insight into insect population variability concerning physiological responses to pollutants. It is indicative that all investigated physiological parameters of L. dispar larvae showed origin-dependent responses to long-term presence of B[a]P, which may be of great importance in ecotoxicological research.

Impact of phenolic compounds on free amino acid contents and electrophoretic patterns of the Nile Tilapia, Oreochromis niloticus

The present research was designed to determine the free amino acids (FAAs) composition and electrophoretic protein patterns by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in Oreochromis niloticus exposed to 1/10 LC50-96 hrs of phenol and nonylphenol ethoxylate 9 (NPEO 9) under laboratory condition for 4 weeks, in an attempt to develop sensitive bio-indicators for evaluation of pollution effects. Variations of FAAs were apparent in both treated groups. Significant increases were recorded in all individual amino acids including essential and non-essential ones, except for proline for both treated groups and leucine for phenol exposed fish group where they decreased significantly. The electrophoretic analysis of protein of the skeletal muscle homogenate using SDS-PAGE indicated that the control group yielded 10 fractions of molecular masses between 166 and 23 kDa. A total number of 9 bands were separated from fish of phenol group; they were varied from 168 to 23 kDa. However, 6 bands only were separated from fish of the NPEO 9 group; they were varied from 157 to 28 kDa. The muscle protein extract from fish of the phenol group showed five unique bands (168, 96, 68, 52, and 39 kDa). Moreover, fish that were exposed to NPEO 9 group showed three unique bands (157, 92, and 72 kDa). The highest similarity index S value observed between phenol and control groups was 0.37, followed by 0.33 between NPEO 9 group and the control one. This could be an indicator of changes in protein metabolism due to phenol and NPEO 9 pollution.

B-RhD-Positive Blood Group Appears Relatively Less Susceptible to Anemia in Sickle Cell Disease Pregnant Women

▪Background: Sickle cell disease in pregnancy accounts for 0.14-0.2% of pregnant women seen in Antenatal clinic in Niger Delta Nigeria.1,2Chronic anemia in SCD during pregnancy could lead to a cascade of maternal and fetal complications. The aim of this study is to assess the various types of ABO blood groups of pregnant SCD women at booking and the severity of anemia. The outcome of this study will help in making recommendations of ABO blood types most suitable to improve the quality of life of SCD patients. This may contribute to guidelines for safe blood transfusion in SCD.Methodology: This was a-ten-year retrospective study of all registered SCD pregnant women seen at the antenatal clinic of Braithwaite Memorial Specialist Hospital (BMSH) Port Harcourt (January 2004- December 2013). Data on gestational age, Hemoglobin (Hb) electrophoretic pattern, Hb concentration and blood group were obtained at booking clinic. Data were analysed using Epi-info version 7.02 by WHO, Geneva Switzerland and CDC, USA.Result: A total of 52 Hb SS pregnant women were seen during the study period. The average booking trimester was the 2nd trimester. 94.2 % (49) were anemic (<11.0 g/dl)3 while 5.8% (3) were non anemic (≥ 11 g/dl). 27.0%, 9.6% and 61.5% were A, B and O rhesus D positive blood groups respectively. No AB blood group was recorded from this study [Table 1]. The O rhesus D positive blood group was significantly more prone to anemia [30 (61.2%)] than other ABO blood groups (X2=50.37; 0.001) [Table 2]. 20% (1/5) of B rhesus D positive and 6.3% (2/32) of O rhesus D positive patients were non-anemic. All A rhesus D positive blood were anemic. The highest prevalence of A rhesus D positive blood group (37.5%) was in 2nd trimester, that of B (20%) and O (66.7%) rhesus D positive blood groups were in 1st and 3rd trimesters respectively [Figure 1].Conclusion: Although most SCD pregnant women in this region are O rhesus D positive blood groups, B rhesus D positive blood group patients appear to be less susceptible to anemia. There is need to carry out more elaborate study on this group of blood.

Bisalbuminemia: A Pathologist's Insight of an Uncommon Phenomenon.

Background The incidence of a bifid electrophoretic pattern in the albumin region on serum protein electrophoresis is an infrequent phenomenon. The availability of literature from India is scarce and is limited to case reports. Objective The aim of the study is to analyze the frequency of bisalbuminemia in an Indian referral facility. The study delved into their clinical associations. Material and Methods The retrospective case records of the patient from the departmental database were scrutinized. The study subjects were for an 8-year study period. Results There were about 39,900 serum electrophoresis performed in an 8-year study period. A total of 40 cases of bisalbuminemia were detected. The incidence in our cohort was 0.01%. Conclusion Bisalbuminemia, an overtly benign condition, is infrequent in Indian population although not rare. It is associated with several clinical disorders; however, the association seems to be plausibly coincidental.

Effect of tear fluid sampling and processing on total protein quantity and electrophoretic pattern.

Human tears contain more than 1500 proteins that could be diagnostically relevant. To date, numerous candidates on a biomarker of protein origin were identified for ocular and systemic diseases. However, the suitable sampling method is still the subject of discussion. To address the need for a description of sampling methods properties for possible clinical analyses, we studied a total protein concentration and electrophoretic pattern of tear fluid collected by capillary tubes, Schirmer strips, cellulose microsponges, and flushing. The total protein concentration was 4.339 μg/μL ± 1.905 μg/μL, 0.967 μg/μL ± 0.117 μg/μL, 0.022 μg/μL ± 0.016 μg/μL, and 0.008 μg/μL ± 0.006 μg/μ for the capillary tubes, Schirmer strips, flushing, and cellulose microsponges, respectively. Sodium dodecyl sulfate polyacrylamide electrophoresis showed the different patterns of tear proteins obtained by the above-mentioned sampling methods. These differences could originate from the use of a bigger amount of extraction reagent that was not used in the case of capillary tubes, and retention of the proteins by strips and sponges. Taken together, capillary tubes, Schirmer strips, cellulose microsponges, and flushing represent sensitive and convenient sampling methods for tear fluid collection. For the isolation of proteins from strips and sponges, and for the flushing, less than 100 μL of a reagent should be used to ensure the sufficient concentration of the biomarkers in a trace amount.

New Bird Sexing Strategy Developed in the Order Psittaciformes Involves Multiple Markers to Avoid Sex Misidentification: Debunked Myth of the Universal DNA Marker.

Sexing of birds is indispensable for scientific, breeding and conservation programs but is difficult in many species and is particularly problematic in the case of nestlings showing no sexual dimorphism. Most useful and efficient methods of sex determination are based on unique features of the Z and W sex chromosomes detected via PCR to distinguish males (ZZ) and females (ZW). During the last twenty-five years researchers searched for the universal marker capable of sexing a maximally wide spectrum of species in a single PCR assay. We screened the phylogenetically representative set of 135 Psittaciformes species including 59 species sexed for the first time. Two known (P2P8, CHD1iA) PCR markers and four additional W/Z polymorphisms (CHD1iE, CHD1i16, CHD1i9 and NIPBLi16) located within the Chromo Helicase DNA binding CHD1 or the Nipped-B homolog NIPBL genes were applied. We present the electrophoretic patterns obtained for the PCR products of the analyzed markers including most typical and atypical patterns allowing sex determination, as well as those obtained when the given marker failed in sexing. Technical aspects of molecular sex determination are discussed: the optimization of amplification conditions, direct PCR and potential misinterpretations. A truly universal marker has not been found, and therefore, we propose a sexing strategy based on multiple CHD1i16, NIPBLi16, CHD1i9 and CHD1iE markers. This new strategy confirms the sex of a given bird with at least two markers detecting independent Z/W polymorphisms, reduces the number of necessary PCR reactions and minimizes the risk of sex misidentification.

Reference intervals for electrophoretograms obtained by capillary electrophoresis of dialyzed urine from healthy dogs.

Electrophoresis of urine to evaluate protein fractions in dogs with proteinuria to differentiate glomerular from tubular damage has increased in recent years; however, capillary electrophoresis (CE) of urine has not been reported in a study of > 40 healthy animals, to our knowledge. We aimed to establish reference intervals (RIs) for the urine protein fractions obtained by CE of urine from healthy dogs. We obtained urine samples from 123 clinically healthy dogs of both sexes between December 2016 and April 2019; urine was frozen until CE was performed. The electrophoretic patterns obtained were divided into 5 protein fractions, and RIs were established in percentages and absolute values using nonparametric methods. RIs were obtained for the fractions (F) as follows: 5.5 to 56.2% for F1, 3.2 to 16.5% for F2, 3.5 to 16.2% for F3, 17.8 to 69.8% for F4, and 5.1 to 23.9% for F5. These RIs obtained by CE might be useful clinically as a basis for comparison with pathologic samples. Age was a statistically significant factor for F2 (p = 0.01) and F3 (p = 0.02), and sex was a statistically significant factor for F1 (p = 0.03).

Effects of cod liver oil fortification on the quality aspects of mince sausages from Indian Major Carp ( Labeo rohita )

Fortification of mince with PUFA-containing oil is an innovative area in product development. In this study, functional sausages were developed from Rohu mince with different concentrations of cod liver oil (CLO) and evaluated for their physiochemical and fatty acid compositions. Fortification of mince sausage with 4% CLO showed highest gel strength (323.58 g.cm) to the product. The CLO product had a higher n-3/n-6 ratio (1.79–3.65 versus 1.19 in control). The electrophoretic pattern of proteins showed the polymerization of proteins in the presence of CLO. The scanning electron microscopy revealed fine texture, absence of more voids, and homogenous structure in 4% CLO-fortified sausages. Hardness values are improved by the addition of CLO with a positive whiteness index and showed better sensory scores during the study period. Therefore, fortification of mince sausages with CLO could improve the health benefits by providing the optimal concentration of essential fatty acids along with good consumer acceptance. Practical applications The mince from Indian Major Carp, Rohu, can be successfully utilized for the development of sausages like ready-to-eat products. The fortification of mince sausages with cod liver oil (CLO) would provide an optimum concentration of essential fatty acid to offer multiple health benefits. The present study was able to achieve a ratio (n-6/n-3) of 0.55 to 0.27 in CLO-fortified sausages, which was considered optimal for human health.

Impact of Extraction Method on the Detection of Quality Biomarkers in Normal vs. DFD Meat.

The objective of this work was to demonstrate how the extraction method affects the reliability of biomarker detection and how this detection depends on the biomarker location within the cell compartment. Different extraction methods were used to study the sarcoplasmic and myofibrillar fractions of the Longissimus thoracis et lumborum muscle of young bulls of the Asturiana de los Valles breed in two quality grades, standard (Control) or dark, firm, and dry (DFD) meat. Protein extractability and the expression of some of the main meat quality biomarkers—oxidative status (lipoperoxidation (LPO) and catalase activity (CAT)), proteome (SDS-PAGE electrophoretic pattern), and cell stress protein (Hsp70)—were analyzed. In the sarcoplasmic fraction, buffers containing Triton X-100 showed significantly higher protein extractability, LPO, and higher intensity of high-molecular-weight protein bands, whereas the TES buffer was more sensitive to distinguishing differences in the protein pattern between the Control and DFD meat. In the myofibrillar fraction, samples extracted with the lysis buffer showed significantly higher protein extractability, whereas samples extracted with the non-denaturing buffer showed higher results for LPO, CAT, and Hsp70, and higher-intensity bands in the electrophoretic pattern. These findings highlight the need for the careful selection of the extraction method used to analyze the different biomarkers considering their cellular location to adapt the extractive process.

Identification, Virulence, and Molecular Characterization of a Recombinant Isolate of Grass Carp Reovirus Genotype I.

The hemorrhagic disease of grass carp (HDGC) caused by grass carp reovirus (GCRV) still poses a great threat to the grass carp industry. Isolation and identification of the GCRV genotype I (GCRV-I) has been rarely reported in the past decade. In this study, a new GCRV was isolated from diseased fish with severe symptoms of enteritis and mild hemorrhages on the body surface. The isolate was further identified by cell culture, transmission electron, indirect immunofluorescence, and SDS-PAGE electrophoretic pattern analysis of genomic RNA. The results were consistent with the new isolate as a GCRV-I member and tentatively named GCRV-GZ1208. Both grass carp and rare minnow infected by the GCRV-GZ1208 have no obvious hemorrhagic symptoms, and the final mortality rate was ≤10%, indicating that it may be a low virulent isolate. GZ1208 possessed highest genomic homology to 873/GCHV (GCRV-I) and golden shiner reovirus (GSRV). Additionally, it was found a 90.7–98.3% nucleotide identity, a 96.4–100% amino acid identity, and <50% identity with GCRV-II and III genotypes. Interestingly, the sequences of some segments of GZ1208 were similar to GCRV-8733/GCHV, whereas the remaining segments were more closely related to GSRV, suggesting that a recombination event had occurred. Bootscan analysis of the complete genomic sequence confirmed this hypothesis, and recombination events between 873/GCHV and other GSRV-like viruses were also accompanied by gene mutations.

Alterations in acute phase proteins, hemogram and electrophoretic pattern of equine serum proteins following change of diet from forage to forage mixed with concentrate

Despite extensive studies on the relationship between diet type and its effect on equine biochemical factors, little is known about horse serum and plasma proteins following changes in diet from forage to forage mixed with concentrate. Acute phase proteins are a group of serum proteins whose concentrations increase in response to infections, inflammation, trauma, and neoplasia. Therefore, in the present study, we evaluated the serum levels of serum biomarkers, fibrinogen and total protein concentration along with the nutritional behavior of horses during diet change. For this purpose, two types of diets, including forage (first treatment) and mixture of forage with concentrate (second treatment) were used for each horse for fifteen consecutive days. Complete blood cell counts as well as serum levels of fibrinogen, haptoglobin and amyloid type A were measured before and after dietary changes. Serum fibrinogen, haptoglobin and amyloid A levels were significantly increased in horses of the second treatment group compared to horses in the first treatment group (p<0.05). There was also a statistically significant difference in the complete blood cell count due to the change of diet from forage to forage mixed with concentrate (p<0.05). It seems that changes in the natural flora of the horse's digestive tract following a change in diet can protect the animal's body against acute inflammatory and traumatic injuries.

Glomalin-related soil protein analysis and its role in erodibility in a semiarid zone in San Luis Potosi, Mexico

The relationship between erodibility (susceptibility to erosion) and soil biological parameters has not been thoroughly investigated in Palomas Valley (PV), a region in San Luis Potosí, México that includes gully-eroded microcatchments. Previous reports have shown the potential of Glomalin-related soil protein (GRSP) as an erodibility marker. Thus, our first goal was to develop an optimized autoclaving-free extraction method for GRSP suitable for 1D-SDS-PAGE analysis. Samples from horizon A (0–10 cm) were collected in three sites in PV. Total GRSP and organic matter contents were obtained, and GRSP was used for correlation analysis (Pearson correlation coefficient) with the K factor, an erodibility index. Our novel total protein-extraction method allowed to obtain moderated resolution electrophoretic patterns and GRSP detection was achieved by molecular weight comparison. GRSP contents ranged 11–203 μg·g−1 soil, and organic matter contents ranged from 2.1 to 12.8%. A multiple regression analysis was performed, obtaining a strong correlation (R = 0.79; R2 = 0.63) among organic matter and GRSP contents and K factor. The results suggested a strong dependence of K from organic components, mainly from GRSP. This is the first time that K factor and GRSP correlation analysis has been reported. Also, this work provides interesting data that could improve the analysis of factor K. All these results suggest GRSP content may play an important role in soil erodibility, but further research is required.

Foliar Application of Auxin or Cytokinin Can Confer Salinity Stress Tolerance in Vicia faba L.

Soil salinity severely declines the availability of water and essential minerals to the plants, which hinders growth. The present study evaluates the potential roles of indole-3-acetic acid (IAA) and 6-benzyladenine (BA) for mitigating the adverse effects of soil-salinity in faba bean (Vicia faba L.). Plants were exposed to 150 mM NaCl stress and were sprayed with IAA (1.15 mM) or BA (0.9 mM). Our results revealed that foliar application of IAA or BA improved the growth traits of salinized faba bean due to the increased uptake of K+, Ca2+, and Mg2+ ions, accumulation of free amino acids, soluble sugars, and soluble proteins, and activity of superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase. The principal component analysis (PCA) and heatmap clustering indicated that salinity-exposed plants exhibited lower growth and biomass production, which correlated with higher accumulation of Na+ and malondialdehyde. Moreover, electrophoretic patterns of protein showed new bands in IAA- or BA-treated salt-stressed plants, indicating that IAA or BA treatment can reprogram the metabolic processes to confer salinity tolerance. We also found that IAA has a greater capacity to ameliorate the salt stress than BA, although there is no significant difference in yield between these treatments. Finally, these findings can be helpful for a better understanding of IAA- and BA-mediated salt tolerance mechanisms and increasing production of faba bean in saline soils.

Assessment of the Ameliorative Effect of Bacillus subtilis against the Toxicity Induced by Aflatoxin B1 in Rats

Aflatoxin B1 (AFB1) is the most toxic and common among the major types of aflatoxins. AFB1 is hepatoxic and has been implicated in an increase risk of hepatocellular carcinoma, also the effect of AFs on the brain chemistry have been documented. Probiotics have health benefits and provide of powerful benefits for the body and brain. The present study was designed to reveal efficiency of Bacillus subtilis NS4182-01 against the hematological, biochemical and histopathological alterations induced by AFB1 in rats. Both of AFB1 and B. subtilis were studied on the experimental animals (rats) that were divided into 6 groups as the following: Group 1 (control) received distilled water orally. Groups 2&3 (B. subtilis treated groups) received B. subtilis orally at a dose of 0.25 and 0.50 ml, respectively. Group 4 (AFB1 treated group) orally treated with AFB1 at a dose of 2 mg/kg b.w. Groups 5&6 (AFB1 + B. subtilis group) orally treated with AFB1 and then treated with B. subtilis at two studied doses respectively. It was noticed that the hematological measurements declined and the most biochemical measurements elevated significantly (P≤0.05) in AFB1 treated group. B. subtilis restored all studied measurements towards the normal values. Moreover, the native electrophoretic protein patterns showed that the physiological alterations occurred in the native protein and lipid moiety in addition to calcium moiety of native protein patterns as a result of AFB1 treatment were represented by hiding one or more of normal protein types with existence of abnormal ones. Therefore, the similarity index (SI%) and genetic distance (GD%) values were altered with protein (SI=40.00%; GD=60.00%), lipid moiety (SI=50.00%; GD=50.00%) and calcium moiety of native protein patterns (SI=75.00%; GD=25.00%) in AFB1 treated group. Although the treatment with B. subtilis at a dose of 0.25 ml showed ameliorative effect but could not restore the physiological state of the patterns to normalcy. While B. subtilis at a dose of 0.50 ml restored integrity of these native protein patterns by restoring the absent types with hiding the abnormal ones. Therefore, this group became physiologically similar to control group (SI=100.00%; GD=0.00%). These findings were supported by results of the histopathological examination in the most target organs (liver, kidney and brain) that were affected by AFB1 and the B. subtilis restored their histopathological integrity to normal structure and maintained architecture of these organs.

Functional modification of protein extracted from black gram by-product: Effect of ultrasonication and micronization techniques

The black gram mill by-product is a valuable protein-rich source that is generally discarded. Therefore, this study was conducted to extract the protein from this underutilized source and further improve its functionality by using ultrasound and micronization techniques. Protein micronized at 110 °C had maximum water absorption (from 2.79 to 4.83 g/g) and emulsifying activity (from 9.94 to 38.32 m2/g). Ultrasonicated proteins for 10 min and 30 min exhibited the highest oil absorption (3.18–3.66 g/g) and foaming capacity (53.7–91.25%), respectively. When compared to ultrasonication, micronization particularly at 150 °C, was very effective in improving zeta potential (−18.7 to −24.95 mV). Thermal properties suggested structural changes, particularly in ultrasonicated proteins. FT-IR spectra showed no marked alteration in the secondary structure, while the electrophoretic pattern showed slight reduction in the intensity of the major band in modified proteins. The findings indicated that these techniques substantially enhanced the functionality of by-product protein, thus might increase its utilization in various food systems.