Extraction, Gene Regulation and Hypocholesterolemic Effects, and Antioxidative Activity of Thymoquinone Rich Fraction and Thymoquinone from Nigella Sativa (pdf incomplete)

Nigella sativa and its active constituent thymoquinone (TQ) have been used for various health benefits such as antitumor, antidiabetic, antihypertensive, antioxidatative and antibacterial. The present study involves thymoquinone rich fraction (TQRF) prepared from Nigella sativa seeds using supercritical fluid extraction (SFE) and commercial available thymoquinone (TQ) to investigate their regulatory effects on genes involved in cholesterol metabolism in vitro using Human liver cancer cells (HepG2 cells) and in vivo using Sprague-Dawley rats. In the present study, SFE extraction procedures with different parameters of pressure at 400 and 600 bar and temperature at 40, 60 and 80°C were optimized to prepare TQRF. The yield of the oil content was highest (37%) at a high temperature of 80°C and a high pressure of 600 bars. The TQ content in different fractions of oil was the highest (2%) at a low temperature of 40°C and a high pressure of 600 bars compared to other used parameters. The major fatty acids in TQRF were oleic and linolenic which exist as unsaturated fatty acids. Palmitic, stearic and myristic acids were the main saturated fatty acids. TQRF was rich in α-tocopherol content (290 ± 1.5 mg/100g). HepG2 cells viability was determined using MTT assay and flow cytrometry. TQRF and TQ were shown to inhibit HepG2 cells growth with IC50 of 100 μg/ml for TQRF and 3.5 μg/ml for TQ after 72 h incubation. Flow cytometry analysis showed that the cells viability was more than 80% when HepG2 cells were treated with TQRF at 50 μg/ml and TQ at 2 μg/ml. The regulatory effects of TQRF at 50 and 80 μg/ml and TQ at 2 μg/ml on genes involved in cholesterol metabolism were studied in HepG2 cell using real time polymerase chain reaction. These genes included low density lipoprotein receptor (LDLR), 3-hydroxy-3-methylglutarylcoenzyme A reductase (HMG-COAR), apolipoproteins (Apo) including Apo A-1 and Apo B100. When cells were treated with TQRF at 50 and 80 μg/ml and TQ at 2 μg/ml mRNA level of LDLR gene was up-regulated by 3, 7 and 2 fold respectively, compared to untreated cells. On the other hand, mRNA level of HMG-COAR was down-regulated by 37, 71 and 12% respectively, compared to untreated cells. The mRNA level of Apo A-1 gene was up-regulated by 3, 4 and 2 fold and Apo B100 was suppressed by 60, 70 and 49% respectively, when cells were treated with TQRF at 50 and 80 μg/ml and TQ at 2 μg/ml compared to untreated cells. TQRF at dose of 0.5, 1 and 1.5 g/kg and TQ at 20, 50 and 100 mg/kg body weight in emulsion form were administrated orally to the rats fed diet supplemented with 1% cholesterol for 8 weeks. There was a significant reduction in plasma total cholesterol levels (TC) and low density lipoprotein cholesterol (LDLC) in both TQRF and TQ at different doses compared to cholesterol control rats (PC). The concentration of the alanine aminotransferase (ALT), gamma glutamyltranspeptidase (GGT), urea and creatinine of plasma collected from experimental rats were also measured in this experiment to test any toxic effect of TQRF and TQ on liver and kidney of rats. ALT, GGT and urea levels were significantly lower in TQRF and TQ treated groups compared to PC group. The antioxidant activity of TQRF and TQ treatment as hydroxyl radical (OH·) scavenging activity in plasma samples collected from experimental rats was also carried out using electron spin resonance (ESR). The findings showed significantly higher of OH· scavenging activity in TQRF and TQ treated rats at different doses compared to untreated rats. The regulatory effect of TQRF and TQ treatment on hepatic genes involved in cholesterol metabolism including LDLR, HMG-COAR, Apo A-1, Apo B100 and Apo E genes were also investigated in vivo. Treating rats with TQRF at dose of 0.5, 1 and 1.5 g/kg and TQ at 20, 50 and 100 mg/kg body weight showed up-regulation of LDLR and Apo E genes whereas both treatments in the same doses showed down-regulation of HMG-COAR and Apo B100 genes compared to untreated rats. In conclusion, the present study suggests that TQRF and TQ generated a hypocholesterolemic effect through regulation of key genes involved in cholesterol metabolism. Both in vitro and in vivo results demonstrate potential value of TQRF and TQ as a novel cholesterol lowering and antioxidant candidate

Nutrient Composition and Hypocholesterolemic Effect of black Seeds (Nigella Sativa L.) and its Oil

Nigella sativa Linn. (Ranuculacea) popularly known as the black seed or Habbatus Sauda is a herb that has traditionally been used for centuries in many parts of the world. It has gained popularity due to its potential health benefits. This study was carried out to investigate the nutrient composition and in vivo hypocholesterolemic effect of the black seeds cultivated in Yemen and its oil. The nutrient composition of three different samples of Nigella sativa seeds from three different regions in Yemen, namely Marib, Sadah and Taiz were studied. Proximate analysis, total dietary fiber (TDF), insoluble dietary fiber (IDF) and soluble dietary fiber (SDF) were determined by the Association of Official Analytical Chemists methods (AOAC). Mineral analysis was carried out using Atomic Absorption spectrophotometer (AAS). Fatty acid methyl esters composition was determined using Gas Chromatography (GC), vitamin E (atocopherol) using HPLC and antioxidant activity by ferric thiocyanate (FTC) and thiobarbituric acid (TBA) methods. Results show that the three samples of Nigella sativa seeds have high content of fat which were 38.4 * 2.1, 37.7 * 0.1, and 36.8 * 1.4% in Sadah, Marib, and Taiz, respectively. In addition, Nigelln sativa seeds have a high content of TDF (36.88 1.44, 26.50 k 1.05, 30.40 * 1.06% for Marib, Sadah and Taiz samples, respectively), IDF (27.10 * 0.55, 20.56 * 1.16, 22.40 * 1.40 % for Marib, Sadah and Taiz samples, respectively) and SDF (8.90 & 1.17, 6.50 * 0.60, 8.13 + 0.71 % for Marib, Sadah and Taiz samples, respectively). Nigella sativa seeds are also rich in calcium, magnesium, potassium, phosphorus and iron. The major fatty acids in Nigella sativa oil extracts from n-hexane were oleic and linolenic which exist as unsaturated fatty acids. Palmitic, stearic and myristic acids were the main saturated fatty acids in all three samples. Oil extracted from the seeds showed higher antioxidant activity compared to a-tocopherol, and was rich in vitamin E (120 + 0.15, 170 k 0.40 and 290 + 1.5 mg/100g for Marib, Sadah and Taiz samples, respectively). The effect of Nigella sativa seed powder and its oil on hypercholesterolemia-induced rabbits was also studied for 8 weeks. A significant reduction (p< 0.05) of total cholesterol (TC) and low density lipoproteins (LDL) in Nigella sativa powder (NSP) and Nigella sativa oil (NSO) groups were observed at weeks 2, 4, 6 and 8. Treatment of rabbits with NSP and NSO showed a significant increase (p>0.05) in plasma HDL levels at weeks 4, 6 and 8 of treatment. A significant decreased (pc0.05) of malondialdehyde (MDA) concentration in erythrocyte and plasma was observed in NSP and NSO groups. There was a significant increase of total plasma antioxidant status (TAS) (p>0.05). NSP and NSO groups also showed significant reduction of serum alanine aminotransferase (ALT) and creatinine concentrations (p<0.05). Plaque formation was significantly inhibited in the aorta, and the tunica intima to tunica media ratio was significantly decreased (p<0.05). In conclusion, this study indicates that the seed of Nigella sativa cultivated in Yemen and its oil contain several nutrients and antioxidants with potential health values such as hypocholesterolemic effect, antioxidant effect and anti atherogenic effect

Backpropagation Neural Network For Colour Recognition

Colour Image Processing (CIP) is useful for inspection system and Automatic Packing Lines Systems. CIP usually needs expensive and special hardware as well as software to extract colour from image. Most of CIP software use statistical methods to extract colours and some system use Neural Network such as Counter-Propagation and Back-Propagation . Some researchers had used Neural Network methods to recognize colour of Commission Internationale de L'Ec1airage (CIE) Models either L *u *v or L *a *b. CIE colour components need special and expensive devices to extract their values from an image. However, this project will use RED, GREEN, BLUE (RGB) colour components, which can be read from an image. In this research, RGB values are used to represent the colour. RGB values are used in two forms. The first form is the actual values that are used in PPM File Format within (0,255) and the second form is normalized RGB values within (0, I ). Back-Propagation Neural Network is used to recognize colour in RGB values. It is found that RGB is useful when used with Neural Network and the Normalized RGB value is faster in the learning of neural network

Thymoquinone rich fraction from Nigella sativa and thymoquinone are cytotoxic towards colon and leukemic carcinoma cell lines.

Nigella sativa has been used for centuries in Asia, Middle East and Africa to promote health and fight diseases. In this study, the anti-cancer effects of thymoquinone rich fraction (TQRF) extracted from N. sativa seeds using supercritical fluid extraction (SFE) system and commercially available thymoquinone (TQ) on colon cancer (HT29), lymphoblastic leukemia (CEMSS) and promyelocytic leukemia (HL60) cells lines were investigated. The concentration that gave 50% inhibition of cell viability (IC50) of HT29, CEMSS and HL60 cells treated with TQRF were 400, 350 and 250 µg/ml, respectively. Meanwhile, the IC50 of TQ was 8, 5 and 3 µg/ml, respectively. Cell cycle analysis shows the increment of apoptosis in a time-dependent manner. However, both TQRF and TQ were not able to arrest the cell cycle phases of the cells. Apoptosis was the main mode of HT29, CEMSS and HL60 cells death induced by both TQRF and TQ. Our findings support the potential use of TQRF and TQ for the treatment of colon cancer and leukemia

Gamma-oryzanol rich fraction regulates the expression of antioxidant and oxidative stress related genes in stressed rat's liver

Background: Gamma-oryzanol (OR), a phytosteryl ferulate mixture extracted from rice bran oil, has a wide spectrum of biological activities in particular, it has antioxidant properties. Methods: The regulatory effect of gamma-oryzanol rich fraction (ORF) extracted and fractionated from rice bran using supercritical fluid extraction (SFE) in comparison with commercially available OR on 14 antioxidant and oxidative stress related genes was determined in rat liver. Rats were subjected to a swimming exercise program for 10 weeks to induce stress and were further treated with either ORF at 125, 250 and 500 mg/kg or OR at 100 mg/kg in emulsion forms for the last 5 weeks of the swimming program being carried out. The GenomeLab Genetic Analysis System (GeXPS) was used to study the multiplex gene expression of the selected genes. Results: Upon comparison of RNA expression levels between the stressed and untreated group (PC) and the unstressed and untreated group (NC), seven genes were found to be down-regulated, while seven genes were up-regulated in PC group compared to NC group. Further treatment of stressed rats with ORF at different doses and OR resulted in up-regulation of 10 genes and down regulation of four genes compared to the PC group. Conclusions: Gamma-oryzanol rich fraction showed potential antioxidant activity greater than OR in the regulation of antioxidants and oxidative stress gene markers

Biofertilizer (EM-1) effect on growth and yield of three bread wheat cultivars

Farmers keep trying to avoid using chemical fertilizer without losing high yield. A field experiment was conducted in the fields of Agriculture College, University of Baghdad during winter seasons of 2015 and 2016 to investigate the response of three bread wheat (Triticumaestivum L.) cultivars (Ibaa99, Abu-Ghraib3 and Buhooth22) to the frequency of spraying with biofertilizer (EM-1) (one time at tillering stage, twice at tillering and stem elongation stages and three times at tillering, stem elongation and booting stages) in addition to the control (without spraying), to the increase of grain yield. Randomized complete block design (RCBD), in split plots arrangement and four replications, was used. Spraying treatments were placed as main plots and cultivars as subplots. The results showed that Ibaa99 cultivar, three times of EM-1 spraying and their interaction gave the highest averages of grain yield (3.89 and 4.31), (3.85 and 4.36) and (4.11 and 4.58 ton*ha-1), respectively, for both seasons. It can be concluded that yield responded significantly to the frequency of EM-1 spraying during vegetative stages

Biofertilizer (EM-1) effect on growth and yield of three bread wheat cultivars

Farmers keep trying to avoid using chemical fertilizer without losing high yield. A field experiment was conducted in the fields of Agriculture College, University of Baghdad during winter seasons of 2015 and 2016 to investigate the response of three bread wheat cultivars (Ibaa99, Abu-Ghraib3 and Buhooth22) to the frequency of spraying with biofertilizer (EM-1) (one time at tillering stage, twice at tillering and stem elongation stages and three times at tillering, stem elongation and booting stages) in addition to the control (without spraying), to the increase of grain yield. Randomized complete block design (RCBD), in split plots arrangement and four replications, was used. Spraying treatments were placed as main plots and cultivars as subplots. The results showed that the Ibaa99 cultivar, three times of EM-1 spraying and their interaction gave the highest averages of grain yield (3.89 and 4.31), (3.85 and 4.36) and (4.11 and 4.58 ton*ha-1), respectively, for both seasons. It can be concluded that yield responded significantly to the frequency of EM-1 spraying during vegetative stages

A Novel Algorithm for Predicting Antimicrobial Resistance in Unequal Groups of Bacterial Isolates

Choosing antimicrobials is a common dilemma when the expected rate of bacterial resistance is high. The observed resistance values in unequal groups of isolates tested for different antimicrobials can be misleading. This can affect the decision to recommend one antibiotic over the other. We analyzed recalled data with the statistical consideration of unequal sample groups. Data was collected concerning children suspected to have typhoid fever at Al Alwyia Pediatric Teaching Hospital in Baghdad, Iraq. The study period extended from September 2021 to September 2022. A novel algorithm was developed to compare the drug sensitivity among unequal numbers of Salmonella typhi (S. Typhi) isolates tested with different antibacterials. According to the proposed algorithm, the predicted resistance values were more valid than the observed values. This proposed algorithm is expected to help the hospital antibiotic policy committee recommend the proper antibacterial agents for S. Typhi and further bacterial isolates

Neonatal cerebral function monitoring – understanding the amplitude integrated EEG

Amplitude integrated electroencephalography (aEEG) is produced by cerebral function monitors (CFM), and is increasingly used in neonates following research into hypothermia for hypoxic ischaemic encephalopathy in term infants. Formal training packages in aEEG in term infants are limited. aEEG is used less often in clinical practice in preterm infants, and requires an understanding of the normal changes seen with increasing gestational age. A number of classifications for aEEG interpretation exist; some purely for term neonates born, and others encompassing both preterm and term neonates. This article reviews the basics of aEEG, its indications and limitations. We also discuss its role in prognostication in term and preterm infants