COMPARATIVE PHYTOCHEMICAL AND BIOLOGICAL INVESTIGATION OF FIVE GLYCINE MAX (L.) MERRILL GENOTYPES

Objective: Soybean (Glycine max L. Merrill) is the world’s most important consumed seed legume. The objectives of the present study were to determine the variability in phytochemical composition and biological activities between five genotypes of G. max. Methods: Lipoidal matters were determined using glucose (GLC). Amino acids were detected by the amino acid analyzer. The phytoconstituents present within each ethanol extract was investigated by gas chromatography-mass spectrometry. The amount of total phenolics, flavonoids, and tannins was analyzed using a spectrophotometric technique, based on Folin–Ciocalteu reagent, aluminum chloride colorimetric assay, and the modified vanillin hydrochloric acid method, respectively. Quercetin, catechin, and gallic acid were used as standard compounds, respectively. Isoflavones content were detected by high-performance liquid chromatography (HPLC)/photodiode array (PDA). The radical scavenging and antioxidant capacity of the genotypes using different in vitro analytical assays such as 2,2-diphenyl,1-picryl hydrazyl, 2,2’-Azino-bis(3-ethylbenzothiazoline-6-sulphonic acid, reducing power, metal chelating, and ferric reducing anti-oxidant power. Butyl hydroxyl toluene and trolox were used as the reference antioxidant radical scavenger compounds. Antitumor activity was evaluated by detecting the viability of Ehrlich ascites carcinoma cells on four different concentrations (1–5 mg/mL). Results: GLC analysis showed the high value of total unsaturated fatty acids and 16 amino acids including glutamic acid with the highest concentration. The variation between genotypes according to their chemical composition of the aldehydes, esters, ketones, alcoholics, and carboxylic content were reported. HPLC/PDA referred to the presence of daidzein, genistein, and in all genotypes. Conclusion: The results confirm the higher value of phytoconstituents of the genotype Giza 35 and Giza 21 as well as their better bioactivity

PREPARATION AND EVALUATION OF CHEMICALLY INACTIVATED SALMONELLA ENTERITIDIS VACCINE IN CHICKENS

  Objective: Salmonella enteritidis ghosts (SEGs) is a non-living empty bacterial cell envelopes which were generated using a different concentration of sodium hydroxide (NaOH) 6.4 mg/mL and evaluated as a vaccine candidate in specific pathogen-free (SPF) chicken. SEGs have been produced by chemical-mediated lysis and evaluated the potential efficacy of chemically induced SEG vaccine and its ability to induce protective immune responses against virulent S. enteritidis challenge in SPF chickens.Methods: SPF chickens were divided into three groups: Group A (non-vaccinated control), Group B (vaccinated with prepared vaccine), and Group C (vaccinated with commercial vaccine).Results: Vaccination of SPF chicken with SEGs induced higher immune responses before and after virulent challenge. SPF chicken vaccinated with SEGs showed increasing in serum enzyme-linked immunosorbent assay (ELISA) antibodies. During the vaccination period, Groups B and C showed higher serum antibody titer compared to Group A. The minimal inhibitory concentration (MIC) of NaOH was capable of inducing non-living SEGs, and it has successfully generated non-living SEGs by MIC of NaOH.Conclusion: It is a one-step process which means easy manufacturing and low production cost compared to protein E-mediated lysis method. Chemically induced SEG vaccine is a highly effective method for inducing protective immunity. This study strongly suggests that SEGs will be a permissive vaccine, as the method of inhibition of S. enteritidis was safe and cheaper than other methods, and it gave a good protection

NEURORESTORATIVE MULBERRIES POTENTIAL OF ALZHEIMER'S DISEASE IN ANIMAL MODEL

Objective: Alzheimer's disease (AD) is one of the most common neurodegenerative disorders. The present study aimed to identify the potential neurorestorative of Morus alba and Morus rubra fruits extract on aluminum chloride (AlCl3)-induced AD rats.Methods: Each Morus species extract M. alba and M. rubra at a dose 300 mg/kg body weight (b.wt.) was orally administered, daily for 6 weeks post AlCl3 induction (100 mg/kg b.wt./day/6 weeks, orally). The polyphenols content was determined according to the colorimetric method of Folin–Ciocalteu using gallic acid as a standard. In addition, brain neurotransmitters, total protein content (TPC), and antioxidant biomarkers were determined. Histopathological assessment was performed on the brains of rats.Results: Administration of AlCl3 neurotoxicity induced a significant increment of brain acetylcholinesterase, and nitric oxide, while significant decrease in four monoamines (norepinephrine, epinephrine, 5-hydroxytryptamine serotonin, and dopamin), along with brain tissue ATPase, and TPC were determined. AlCl3-induced changes in peroxidative products, such as protein carbonyl, catalase, and superoxide dismutase activity. AlCl3 induction resulted in significant decrease in antioxidant enzymes, serum level of total antioxidant capacity, and caused histological alteration for brain tissue. Treatment of AD-rats with M. alba and M. rubra (300 mg/kg b.wt./day) causes modulation of most biochemical markers brain function and confirmed by brain histopathological study.Conclusion: Collectively, the present results indicated that M. rubra and M. alba exerts neuroprotection against AlCl3-induced brain pathology. To the best of our knowledge, none of the literature studies have demonstrated the effect of these Morus species against Al-neurotoxicity at biochemical and electron microscopic levels

CHEMICAL COMPOSITION AND BIOLOGICAL ACTIVITIES OF TWO SOLANUM TUBEROSUM CULTIVARS GROWN IN EGYPT

Objective: This paper reports a comparative study based on the chemical composition, antioxidant and therapeutic effect of two Egyptian cultivars of Solanum tuberosum; Baraka (STB) and Alpha (STA) on hepatic functions and oxidative stress in alloxan (ALX)-treated rats.Methods: The potential of cultivars as a source of natural antioxidants were explored using five in vitro assays, and the results were compared with butylated hydroxyl toluene and Trolox. The modulator role of cultivars was assessed by determining its effect on oxidative stress measured by lipid peroxide (LPO) levels, serum aminotransferases and on antioxidative enzymatic activities of glutathione transferases, malondialdehyde, nitric oxide, catalase, superoxide dismutase, and glutathione peroxidase. In addition, histological examinations of liver, kidney, pancreas and spleen were carried out to confirm the biochemical changes of the diabetic group of rats non-treated and treated with STB, STA and glibenclamide as standard drug.Results: Our findings demonstrated variations in antioxidant activities of each cultivar. An increase in LPO, aminotransferases levels was observed in the ALX-induced diabetic rats. Administration of each cultivar for four w caused a significant reduction in LPO, serum aminotransferases levels in the ALX-induced diabetic rats. Furthermore, a significant improvement in the activities of all the investigated antioxidant enzymes was marked. In addition, the total content of phenolics, flavonoids, anthocyanins, tannins, and alkaloids were varied in each cultivar.Conclusion: STB evidenced remarkable bioactivity compared to that exhibited by STA. Moreover, three known compounds, previously not isolated from Solanum genus, were obtained from the methylene chloride fraction of STB.Â

PHYTOCHEMICAL ANALYSIS, ASSESSMENT OF ANTIPROLIFERATIVE AND FREE RADICAL SCAVENGING ACTIVITY OF MORUS ALBA AND MORUS RUBRA FRUITS

Objective: Mulberry is a nontoxic commonly eaten plant, belongs to the Morus and used in folk medicine in the remedy of dysentery, antiphlogistic, diuretic, expectorant, and antidiabetic. The purpose of this study is to evaluate the antiproliferative and radical scavenging activity of the total alcoholic and successive fractions thereof of Morus alba and Morus rubra fruits. In addition, the chemical composition of the bioactive fractions of each species was investigated.Methods: The antiproliferative potential of 8 extracts on 4 human cancer cell lines, hepatocellular carcinoma (HepG2), Caucasian breast adenocarcinoma (MCF7), prostate (PC3), and colon carcinoma (HCT116) in addition to one normal cell line namely human normal immortalized skin fibroblast cells (BJ1) were carried out. Cell viability was determined using MTT assay. The potency was compared with the reference drug doxorubicin. These extracts were also assayed for 1,1-diphenyl-2-hydrazyl free radical scavenging activities. After saponification of the n-hexane fraction, unsaponifiable matter and fatty acid methyl esters were analyzed by gas liquid chromatography (GLC). The chemical composition of the bioactive fractions was investigated using gas chromatography/mass spectrometry (GC/MS) analysis.Results: All the extracts showed significant free radical scavenging activity dose-dependently. The n-hexane and dichloromethane (DCM) fractions of M. rubra exhibited potent cytotoxic activity on almost cancer cell lines. In the same pattern, ethyl acetate (EtOAc) of M. rubra has moderate cytotoxic activity against all cell lines except HepG2. DCM fraction of M. alba possessed both radical scavenging and high potential antiproliferated activities against HCT116 and MCF7 with inhibitory concentration of 43.9 and 32.3 μg/ml, respectively, while it showed no cytotoxic effect on BJ1. GLC analysis showed the major hydrocarbons in M. alba and M. rubra were heptacosane and docosane, respectively. Sterols were similar in both species but with different ratios and cholesterol was the major one. Palmitic and margaric were the major saturated fatty acid while arachidonic was the major unsaturated fatty acid in both species. GC/MS analysis showed the main compound in DCM fraction of each Morus species was palmitic acid. Furthermore, 1,11-bis-(methoxycarbonyl-ethenyl)-10,2-dihydroxy-cycloeicosane and linolelaidic acid, methyl ester were the main compounds in the EtOAc fraction of each Morus species. Whereas, the main compounds in alcoholic extract of M. alba and M. rubra were methyl-14-methyl-pentadecanoate and 1,2-O-isopropylyidene-4-nonene-1,2,3-triol, respectively.Conclusions: The results observed remarkable biological activity of the successive fractions of M. rubra more than those of M. alba and confirmed its importance as a natural bioactive source. Morus species are good candidates to be promising as possible sources for future antitumor and antioxidants in food and pharmaceutical formulations. The strong activity partly explains the potential effects of Morus species for the treatment of cancer and degenerative diseases caused by free radicals

BIOINFORMATICS: INFLAMMATORY CYTOKINES AND ATTENUATION OF DIABETES HYPERCHOLESTEROLEMIA-INDUCED RENAL INJURY USING MORNING GLORY AND NECKLACE POD EXTRACTS

  Objective: The present research in bioinformatics focuses on pharmacological effects of morning glory and necklace pod ethanolic extracts (MGE and NPE) on some biochemical parameters in high fat diet-induced hypercholesterolemia and streptozotocin-induced hyperglycemia in rats.Methods: Compared to atorvastatin; an anti-hypercholesterolemic (HC) and glibenclamide; an antidiabetic drug. Endothelium activation markers of soluble vascular cell adhesion molecule (VCAM)-1 and intercellular adhesion molecule (ICAM)-1 were determined using enzyme-linked immunosorbent assay. Creatinine, urea, and inflammatory biomarkers; C-reactive protein (CRP) and pro-inflammatory cytokines including tumor necrosis factor alpha (TNF-α) and interleukin (IL)-10 levels were also measured in serum of different therapeutic groups.Results: Significant decrease in ICAM-1 level with MGE and NPE supplemented to normal rats as compared to untreated control with percentages decrease 17.80 and 12.00% was observed. Insignificant change was detected in VCAM-1 level. Profound amelioration in CRP, total urea and creatinine levels by NPE treatment. Creatinine, urea, CRP, and TNF-α level were significantly increased in hyperglycemic (HG)-HC rats. However, IL-10 level showed a significant decrease. Meanwhile, histopathological investigation of the kidney and heart was carried out. Image recognition system for kidney and heart images was developed to diagnose their diseases. Tested extract attenuated creatinine, urea, CRP, and TNF-α level. Hyperglycemia and hypercholesterolemia linked kidney disorders were relieved.Conclusion: In vivo oral administration with each extract declared suppression of cytokines mediated inflammation, vascular function leading to infiltration reduction of renal macrophage together with lowering in kidney indices and ameliorate renal tissues architectures in HG-HC rats

PHYTOPHENOLICS COMPOSITION, HYPOLIPIDEMIC, HYPOGLYCEMIC AND ANTIOXIDATIVE EFFECTS OF THE LEAVES OF FORTUNELLA JAPONICA (THUNB.) SWINGLE

Objective: Fortunella japonica (Thunb.) Swingle is an evergreen shrub, its whole fruit, including the peel, is eaten. There have been few detailed phytophenolics composition reports on this genus and the hypoglycemic and hypolipidemic effects of the plant were not evaluated. Methods: Structures of the isolated compounds were elucidated by spectral analysis. Serum glucose level, activities of liver enzymes, total protein content, serum lipid profiles, antioxidant parameters and some glycolytic and gluconeogenic enzymes in streptozotocin (STZ)-induced diabetic rats were determined. The evaluation also carried out through determination of liver disorder biomarkers and histopathological examination of liver, kidney and pancreas. Results: Six phytophenolics were isolated, for the first time from the genus Fortunella as well as a sterol compound. Treatment with the ethanolic extract of F. japonica leaves effectively meliorated antioxidant markers and glycolytic enzymes. The histopathological analyzes also confirmed the experimental findings.Conclusion: The results show that the ethanolic extract has hypoglycemic, hypotriglyceridemic and antioxidant effects in STZ-induced diabetic rats, suggesting that this extract supplementation can be useful in preventing diabetic complications associated with hyperlipidemia and oxidative stress.Â

STATISTICAL DATA ANALYSIS WHICH RESULT FROM THE BIO-DIAGNOSIS AND BIO-TREATMENT OF INJURED RATS WITH THE HYPERLIPIDEMIA AND HYPERGLYCEMIA DISEASES

ABSTRACTObjective: This study in bioinformatics aims to investigate the potential effect of Ipomoea tricolor and Sophora tomentosa on liver function enzymesactivity, serum lipid profile, oxidative stress biomarkers, and on blood glucose in high fat diet-induced hypercholesterolemia (HC) and STZ-inducedhyperglycemia (HG) in rats.Methods: Blood glucose level, liver function enzymes, alanine aminotransferases and aspartate aminotransferases, alkaline phosphatase, and lactatedehydrogenase (LDH) were determined. Besides, lipid profile including total cholesterol (TC), triacylglycerol (TG), total lipid, and high-densitylipoprotein-cholesterol was investigated. Moreover, oxidative stress biomarkers, lipid peroxide, and nitric oxide as well as non-enzymatic antioxidant,glutathione (GSH) were also examined in different therapeutic groups.Results: A significant increase in blood glucose level, liver function enzyme activities, LDH, lipid profile and oxidative stress markers, while significantdecrease in LDH-C and GSH level in HC-HG induced rats compared to control one. A marked amelioration in all biochemical parameters underinvestigation on treatment of HC-HG rats with I. tricolor and S. tomentosa with different fluctuating percentages of improvement. Histopathologicalexamination of liver and pancreas was also performed and declared HC-HG showed congestion in portal vessels and sinusoids with mild centrilobularhepatocyte degeneration, marked hepatocyte ballooning and hydropic degeneration, while HC-HG treated rats with I. tricolor and S. tomentosa showednormal lobular hepatic architecture with mild sinusoidal dilatation and congestion. On the other hand, a histological organization of pancreas of HC-HGrats showing disarrangement changes in pancreatic blood vessels and interlobular duct as well as disordered in acini. The treatment of HC-HG rats withI. tricolor and S. tomentosa showed enhancement in Langerhans cells and restore of most pancreatic tissue in comparison with standard drugs.Conclusion: The statistical results showed that each extract ameliorated high blood glucose level liver injury, HC and oxidative stress indicatingrelieving of oxidative damage associated with the complexity of HG and HC. These results demonstrated that these two plants extracts may be acandidate intelligent antioxidant, hypolipidemic, hypoglycemic, and hepatoprotective nutraceuticals which need further clinical investigation to beapplied effectively to reduce perturbation in HC associated diabetes.Keywords: Ipomoea tricolor, Sophora tomentosa, Lipid profile and liver function enzymes, Endothelial dysfunction markers, Statistics and imagerecognition, Histopathological analysis

SYNTHESIS AND ANTIVIRAL ACTIVITY OF NOVEL ETHYL 2-(3-HETEROCYCLE-1H-INDOL-1-YL) ACETATE DERIVATIVES

Objective: Marek's disease (MD) is a widespread, herpesvirus-induced neoplastic disease in the domestic chicken that is caused by Marek's disease virus (MDV). Marek’s disease virus (MDV) belongs to the alphaherpesvirus family such as Herpes simplex viruses 1 and 2 (HSV-1 and HSV-2). Recently Bag and co-workers 2014 reported that, 7-methoxy-1-methyl-4, 9-dihydro-3H-pyrido [3, 4-b]indole (Harmaline) showed potent anti-HSV-1 activity against both wild type and clinical isolates of HSV-1. The present work aimed to synthesize some new heterocyclic systems incorporated to indole moiety starting from ethyl 2-(3-acetyl-1H-indol-1-yl)acetate (1) in order to evaluate their antiviral activity in a trail to explore potential antiviral agents against MDV to limit the disease course and losses. Methods: Reaction of ethyl 2-(3-acetyl-1H-indol-1-yl) acetate (1) with semicarbazide hydrochloride yielded semicarbazone derivative 2. The oxidative cyclization of 2 using thionyl chloride and selenium dioxide afforded 1, 2, 3-thia and 1, 2, 3-selenadiazole derivatives 3 and 4, respectively. On the other hand, reaction of 1 with 4-chloro and 4-nitrobenzaldehydes under Claisen-Schmidt conditions gave α, β-unsaturated keto derivatives 5a, b. Cyclization of 5a, b using hydrazine hydrate, phenyl hydrazine, urea, thiourea or guanidine led to the formation of pyrazoles 6a, b, 7a, b, and pyrimidines derivatives 8a, b-10a, b; respectively. Condensation of 1 with phenyl hydrazine followed by Vilsmeier Haack formylation gave pyrazole-4-carboxaldehyde derivative 12. Reaction of aldehydic function group of 12 with different reagents led to the formation of pyrazol-5-ones 14-16, thiazolidinone 18, aziditine 19, 1, 6-diaminopyridine 21, triazolo(1, 5-a)pyridine 22 and pyrano(2, 3-c) pyrazole derivatives 23. The in vitro antiviral activity of the selected compounds 6a, b 7a, b 8a, b 9a, b and 10a, b was studied against Marek's disease virus (MDV). Results: Chicken embryo experiment showed that compounds 7b, 8b, 9b and 10a possessed significant antiviral activity with IC50 ranged between 5 and 6 µg/ml and substantial therapeutic indices (TI) of 80 and 83 were recorded. Cytotoxicity assay indicated that CC50 of 7b, 8b, 9b and 10 were greater than 400 and 500 mg/ml. Conclusion: Compounds 7b, 8b, 9b and 10a showed promising effect as anti-MDV infectivity application

BERRIES SUPPLEMENTATION MODULATES BODY WEIGHT AND METABOLIC DETERIORATIONS IN OBESE RATS

 Objective: The aim of the present work is to investigate the potential of purple (Morus rubra [MR]) and white (Morus alba [MA]) berry supplementation to modulate body weight (BW) and metabolic deteriorations in obese Wistar male rats.Methods: Seventy rats weighing 150.00±10 g (mean±standard deviation) were used; the rats were randomly divided into seven groups of 10 rats each. Obesity was induced by feeding animals high-fat diet (HFD), for 6 consecutive weeks then treated with the purple (MR) and white (MA) berry ethanol extracts for 6 weeks in a dose 300 mg/Kg BW. BW gain, food intake fecal fat content, food consumptions were determined. The fats and organs such as liver, kidney, and heart were also weighed. Lipid profiles included triglyceride, total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol, as well as liverenzymes; alanine aminotransferase, alkaline phosphatase, aspartate aminotransferase, γ-glutamyltransferase (γ-GGT), and leptin were also determined.Results: Treatment of obese rats with white and purple berries showed significant reduction in BW reached to 23.68% with MR and 21.19% with MA berries compared to Orlistat (31.16%). Liver weight, visceral fat, and liver enzyme activities were also markedly ameliorated. In addition, lipid profile biomarkers were improved on treated obese rats with both Morus species. In addition, increase in fecal fat (250.73% and 256.25%, for MR and MA, respectively) was noticed while blood glucose, insulin, leptin levels, and pancreatic lipase activity was improved. In addition, supplementing obese rats with both Morus species extract significantly increase food intake reached 33.33% and 36.66%, for MR and MA, respectively.Conclusion: The ameliorating effect of both Morus extracts may be attributed to polyphenolic rich compounds which have antioxidant properties, suppressed lipid synthesis in hepatic tissue and inhibited pancreatic lipase activity which consequently recommended using as a promising anti-obesity agents