Zeaxanthin Isolated from Dunaliella salina Microalgae Ameliorates Age Associated Cardiac Dysfunction in Rats through Stimulation of Retinoid Receptors.

Retinoids are essential during early cardiovascular morphogenesis. However, recent studies showed their important role in cardiac remodeling in rats with hypertension and following myocardial infarction. The present study aimed to investigate the effect of zeaxanthin heneicosylate (ZH); a carotenoid ester isolated from Dunaliella salina microalgae, on cardiac dysfunction ensuing d-galactose injection in rats. Rats injected with d-GAL (200 mg/kg; I.P) for 8 weeks were orally treated with ZH (250 µg/kg) for 28 consecutive days. Results showed that d-GAL injection caused dramatic electrocardiographic changes as well as marked elevation in serum levels of homocysteine, creatinine kinase isoenzyme and lactate dehydrogenase. A reduction in the cardiac contents of glucose transporter-4 and superoxide dismutase along with the elevation of inducible nitric oxide synthetase and interleukin-6 was also noticed. Oral administration of ZH significantly improved the above mentioned cardiac aging manifestations; this was further emphasized through histopathological examinations. The effect of ZH is mediated through the interaction with retinoid receptor alpha (RAR-α) as evidenced through a significant elevation of RAR-α expression in cardiac tissue following the lead of an in silico molecular docking study. In conclusion, zeaxanthin heneicosylate isolated from D. salina ameliorated age-associated cardiac dysfunction in rats through the activation of retinoid receptors.

Cytotoxic activity of carotenoid rich fractions from Haematococcus pluvialis and Dunaliella salina microalgae and the identification of the phytoconstituents using LC-DAD/ESI-MS.

Microalgae represent a rich source that satisfies the growing need for novel ingredients of nutriceuticals, pharmaceuticals, and food supplements. Haematococcus pluvialis and Dunaliella salina microalgae are isolated from the Egyptian hydro-flora and are reported for their potent antioxidant activities. The cytotoxic activity of different fractions of both microalgae was investigated on 4 cell lines HePG2, MCF7, HCT116, and A549. The carotenoid rich fraction of H. pluvialis showed potent cytotoxic activity against colon cancer cell line and moderate activity against both liver and breast cancer cell lines. On the other hand, the carotenoid rich fraction of D. salina showed mild cytotoxic activity on breast and liver cancer cell lines. The carotenoid rich fraction of H. pluvialis was analysed using LC-DAD/ESI-MS and the major carotenoids were identified either free as well as bounded to fatty acids.

Dietary supplementation with Dunaliella salina microalga promotes quail growth by altering lipid profile and immunity.

The goals of the current research are to ascertain the impacts of Dunaliella salina (DS) on quail growth, carcass criteria, liver and kidney functions, lipid profile, and immune response. Two hundred and forty 7-day-old quail chicks were divided equally into 4 separate groups with 6 replicates with 10 birds each. The groups were as follows: 1) control diet (the basal feed without DS), 2) control diet enriched with 0.25 g DS/kg, 3) control diet enriched with 0.50 g DS/kg, and 4) control diet enriched with 1.00 g DS/kg. Results elucidated that the birds which consumed 0.5 and 1 g DS/kg diet performed better than other birds in terms of live body weight (LBW), body weight gain (BWG), and feed conversion ratio (FCR). There were no significant changes in feed intake (FI) and carcass characteristics due to different dietary DS levels. Compared to the control group, DS-treated groups had better lipid profile (low total cholesterol and LDL values and high HDL values) and immune response (complement 3 values). The quails consumed feeds with different levels of DS had greater (P < 0.038) C3 compared to control. Adding 0.5 and 1 g DS/kg lowered blood concentrations of triglycerides and total protein (TP) values. The high level of DS (1 g/kg) had higher albumin values and lower AST values than other groups (P < 0.05). The creatinine values were at the lowest levels in the group consumed 0.50 g DS/kg feed. No changes (P > 0.05) were demonstrated among experimental groups in the ALT, urea, and lysozyme values. In conclusion, adding D. salina to growing quail diets enhanced growth, immune system, blood lipid profile, and kidney and liver function.

Natural ß-carotene prevents acute lung injury induced by cyclophosphamide in mice.

IL-17 is associated with varied inflammatory and immune-related diseases. However, the biological function of IL-17 and its expression in acute lung damage are not entirely known. Thanks to the powerful antioxidant properties of ß-carotene, we presumed that it would show a potent protecting effect against cyclophosphamide (CP) -induced acute lung injury (ALI) in mice. We studied the mechanisms underlying the effect of ß-carotene supplementation against CP-induced ALI in mice. We isolated the ß-carotene from Scenedesmus obliquus microalgae n-hexane extract and identified it by HPLC and 1H-NMR analysis. Within the experiments, 40 mice were assigned into five groups randomly: Group 1 (Control): Mice received saline. Group 2 (ß-carotene control): Mice were administered ß-carotene (40 mg/kg; orally) once daily for 10 sequent days without CP injection. Group 3 (CP): One i.p injection of 200 (mg/kg) of CP was given to mice. Group 4 and 5 (CP + ß-carotene): Mice were administered ß-carotene (20 and 40 mg/kg; orally) once a day for ten days following the CP injection. Lung samples were collected for lab analysis, after scarifying the animals at the experiment end. Administration of ß-carotene orally reduced CP-induced ALI and inflammation. ß-carotene significantly decreased wet-to-dry weight ratios (W/D), down-regulated IL-17, NF-κB, and IKBKB, decreased the contents of TNF-α, COX-2, and PKC, and increased the contents of SIRT1 and PPARγ in the lung tissues. ß-carotene ameliorated the histopathological changes induced by CP and reduced the scoring number of inflammatory cell infiltration and emphysema when compared to CP. Consequently, we conclude natural ß-carotene is a promising anti-inflammatory mediator for different inflammatory-related complications.

Lutein isolated from Scenedesmus obliquus microalga boosts immunity against cyclophosphamide-induced brain injury in rats.

Lutein is a naturally potent antioxidant carotenoid synthesized in green microalgae with a potent ability to prevent different human chronic conditions. To date, there are no reports of the immune-stimulating effect of pure lutein isolated from Scenedesmus obliquus. Thus, we isolated the natural lutein from S. obliquus and evaluated its effectiveness as an immunostimulant against cyclophosphamide-induced brain injury. We purified all-E-(3R, 3'R, 6'R)-Lutein from S. obliquus using prep-HPLC and characterized it by 1H- and 13C-NMR spectroscopy. We assigned rats randomly to four experimental groups: the Control group got a vehicle for lutein dimethyl sulfoxide for ten successive days. The Cyclophosphamide group received a single i.p injection of Cyclophosphamide (200 mg/kg). Lutein groups received 50 and 100 (mg/kg) of lutein one time per day for ten successive days after the cyclophosphamide dose. Lutein administration reduced brain contents of Macrophage inflammatory protein2 (MIP2), cytokine-induced- neutrophil chemoattractant (CINC), and Matrix metalloproteinase 1 (MMP1). Besides, it lowered the contents of interleukin 1 beta (IL-1ß) and interleukin 18 (IL-18), associated with low content of NLR pyrin domain protein 3 (NLRP3) and consequently caspase-1 compared to the cyclophosphamide group. In the histomorphometric analysis, lutein groups (50 and 100 mg/Kg) showed mild histopathological alterations as they significantly reduced nuclear pyknosis numbers by 65% and 69% respectively, compared to the cyclophosphamide group. This is the first study that showed the immunomodulatory roles of lutein against cyclophosphamide-induced brain injury via decreasing neuroinflammation, chemokines recruitment, and neuron degeneration with the modulation of immune markers. Hence, lutein can be an effective immunomodulator against inflammation-related immune disorders.

Amelioration of Hepatic Encephalopathy Using Dunaliella salina Microalgae in Rats: Modulation of Hyperammonemia/TLR4.

Hepatic encephalopathy (HE) is a neuropsychiatric disease that is developed as a complication of both acute and chronic liver failure affecting psychomotor dysfunction, memory, and concentration. This study is aimed at evaluating the therapeutic effects of Dunaliella salina (D. salina) microalgae in thioacetamide- (TAA-) induced HE in rats. HE was induced by TAA (200 mg/kg; i.p.) for three successive days. Forty male Wister albino rats were divided into 4 groups; the first group was served as a normal, and the second group was injected with TAA and served as TAA control. The third and fourth groups were administered D. salina (100 and 200 mg/kg; p.o.), respectively, after TAA injection for 7 days. The behavioral and biochemical markers as well as histological aspects of HE were estimated. This study revealed that TAA caused behavioral changes, oxidative stress, neuroinflammation, nuclear pyknosis, and neurons degeneration. D. salina improved liver function and decreased oxidative stress and inflammatory mediator as TLR4 protein expression. Also, D. salina elevated HSP-25 and IGF-1 as well as improved brain histopathological alterations. In conclusion, D. salina exerted a therapeutic potential against HE via its antioxidant, antiinflammatory and cytoprotective effects.

Haematococcus pluvialis Carotenoids Enrich Fractions Ameliorate Liver Fibrosis Induced by Thioacetamide in Rats: Modulation of Metalloproteinase and Its Inhibitor.

Hepatic fibrosis is a consequence of chronic liver diseases. Metalloproteinase and its inhibitor have crucial roles in the resolution of liver fibrosis. The current relevant study is aimed to evaluate the therapeutic effect of Haematococcus pluvialis (H. pluvialis) extract, astaxanthin-rich fraction, astaxanthin ester-rich fraction, and ß-carotene-rich fraction as well as their mechanisms of action in curing hepatic fibrosis induced by thioacetamide (TAA). Liver fibrosis was induced using TAA (intraperitoneal injection, two times a week for 6 weeks), in a rat model and H. pluvialis extract (200 mg/kg), and other fractions (30 mg/kg) were orally administered daily for 4 weeks after the last TAA injection. Based on HPLC analysis, H. pluvialis extract contains ß-carotene (12.95 mg/g, extract) and free astaxanthin (10.85 mg/g, extract), while HPLC/ESI-MS analysis revealed that H. pluvialis extract contains 28 carotenoid compounds including three isomers of free astaxanthin, α or ß-carotene, lutein, 14 astaxanthin mono-esters, 5 astaxanthin di-esters, and other carotenoids. H. pluvialis and its fractions reduced liver enzymes, nitric oxide, collagen 1, alpha-smooth muscle actin, and transforming growth factor-beta as well as elevated catalase antioxidant activity compared to the TAA group. Also, H. pluvialis extract and its fractions exceedingly controlled the balance between metalloproteinase and its inhibitor, activated Kupffer cells proliferation, and suppressed liver apoptosis, necrobiosis, and fibrosis. These findings conclude that H. pluvialis extract and its fractions have an antifibrotic effect against TAA-induced liver fibrosis by regulating the oxidative stress and proinflammatory mediators, suppressing multiple profibrogenic factors, and modulating the metalloproteinase and its inhibitor pathway, recommending H. pluvialis extract and its fractions for the development of new effective medicine for treating hepatic fibrosis disorders.

Dunalialla salina microalgea and its isolated zeaxanthin mitigate age-related dementia in rats: Modulation of neurotransmission and amyloid-ß protein.

Age-related deterioration of sensorimotor and cognitive abilities suggests that the brain undergoes regressive alterations with aging that compromise its function. Thus, the present study was designed to assess the efficacy of Dunaliella salina in counteracting D-galactose (D-gal)-induced dementia brain aging and its modulatory role in attenuating amyloid ß (Aß) protein and neurotransmitters. Aging associated dementia was generated by injection of D-gal (200 mg/kg; i.p) of rats for 8 weeks. D. salina biomass (250 mg/kg), polar (30 mg/kg), its carotenoid (30 mg/kg) fractions as well as the isolated zeaxanthin (250 µg/kg) were given orally simultaneously with D-gal for additional two weeks. Twenty-four hours after the last treatment dose; behavioral, biochemical and histopathological assessment were performed. Results showed that oral treatment of motor deficit rats with D. salina biomass and its isolated polar and carotenoid fractions showed amelioration in the motor coordination assessed by the rotarod test and in the memory and learning capabilities evaluated by Morris water maze test. D. salina also showed a reduction in brain levels of inflammatory indicators viz. interlekin-1ß and inducible nitric oxide synthetase as well as brain contents of Aß protein and myelin base protein. Likewise, oral treatment with D. salina biomass and its isolated polar and carotenoid fractions exhibited an increase in the rats' brain neurotransmitters and their metabolites. Furthermore, histopathological investigations have confirmed all of these results. Our findings suggest that D. salina overcomes brain aging and thereby repairs age-related dementia, both for its modulating function in attenuating the Aß protein and neurotransmitters.

Enhancing antioxidant availability in wheat grains from plants grown under seawater stress in response to microalgae extract treatments.

BACKGROUND: The present study was designed to investigate the enhanced antioxidant capacity of whole grains from wheat plants grown under seawater stress in response to microalgae extract treatment. RESULTS: The total carotenoid (TCAR), tocopherol (TOC), phenolic (TPC), and protein (PC) contents in whole grains of wheat plants irrigated with 10% and 20% (v/v) seawater (SW) in response to water extracts of microalgae Spirulina maxima (SME) and Chlorella ellipsoida (CEE) as well as exogenous plant growth enhancers of ascorbic acid and benzyladenine treatments were measured. The results showed that the levels of enhanced TCAR (range 0.08-0.14 g kg(-1)), TOC (range 0.05-0.12 g kg(-1)), TPC (range 0.80-2.96 g kg(-1)) and PC (range 93.4-137.9 g kg(-1)) in wheat grains of plants irrigated with 10% and 20% SW were significantly increased in response to SME and CEE treatments. Evaluation of antioxidant activity of ethanolic extracts of grains of SW-stressed plants indicated that DPPH and TBAS radical scavenging activity was significantly increased in response to SME and CEE treatments and coincided with the increase in levels of antioxidant compounds present in each extract. The electrophoretic profiles of the grains of proteins of treated samples exhibited quite different patterns from those in control samples. CONCLUSION: The results suggest that the application of algal extracts to wheat plants irrigated with SW is useful for improvement of salinity tolerance. This effect can be triggered by the stimulation of antioxidant components and protein content.

Dunaliella salina Attenuates Diabetic Neuropathy Induced by STZ in Rats: Involvement of Thioredoxin.

Diabetic neuropathy (DN) is a widespread disabling disorder including peripheral nerves' damage. The aim of the current study was to estimate the potential ameliorative effect of Dunaliella salina (D. salina) on DN and the involvement of the thioredoxin. Diabetes was induced by streptozotocin (STZ; 50 mg/kg; i.p). Glimepiride (0.5 mg/kg) or D. salina powder (100 or 200 mg/kg) were given orally, after 2 days of STZ injection for 4 weeks. Glucose, total antioxidant capacity (TAC), superoxide dismutase (SOD), and catalase (CAT) serum levels as well as brain contents of thioredoxin (Trx), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6) were measured with the histopathological study. STZ-induced DN resulted in a significant (P < 0.05) rise in glucose blood level and brain contents of TNF-α and IL-6 and produced a reduction in serum TAC, SOD, CAT, and brain Trx levels with irregular islets of Langerhans cells and loss of brain Purkinje cells. Treatment with glimepiride or both doses of D. salina alleviated these biochemical and histological parameters as compared to the STZ group. D. salina has a neurotherapeutic effect against DN via its inhibitory effect on inflammatory mediators and oxidative stress molecules with its upregulation of Trx activity.

Dunaliella salina microalgae oppose thioacetamide-induced hepatic fibrosis in rats.

Several hepatic pathological conditions are correlated with the stimulation of hepatic stellate cells. This induces a cascade of events producing accretion of extracellular matrix components triggering fibrosis. Dunaliella salina, rich in carotenoids, was investigated for its potential antagonizing activity; functionally and structurally against thioacetamide (TAA) - induced hepatic fibrosis in rats. Adult male albino Wistar rats were treated with three dose levels of D. salina powder or extract (daily, p.o.); for 6 weeks, concomitantly with TAA injection. Serum levels of aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), bilirubin and albumin were determined. Reduced glutathione (GSH), malondialdehyde (MDA), smooth muscle actin alpha (α-SMA) and collagen I hepatic contents were also estimated. Treatment with D. salina powder or extract caused a significant decline in serum levels of AST, ALT, ALP, bilirubin, MDA and hepatic contents of α-SMA and collagen I. Additionally, serum albumin and GSH hepatic content were highly elevated. Liver histopathological examination also indicated that D. salina reduced fibrosis, centrilobular necrosis, and inflammatory cell infiltration evoked by TAA. The results implied that D. salina exerts protective action against TAA-induced hepatic fibrosis in rats. The phytochemical investigation revealed high total carotenoid content prominently ß-carotene (15.2 % of the algal extract) as well as unsaturated fatty acids as alpha-linolenic acid which accounts for the hepatoprotective activity.

Attenuation of Age-Related Hepatic Steatosis by Dunaliella salina Microalgae in Senescence Rats through the Regulation of Redox Status, Inflammatory Indices, and Apoptotic Biomarkers.

BACKGROUND: Hepatic steatosis is the most common type of chronic liver disease and is considered an established risk factor of major chronic diseases. PURPOSE: The present study aimed to investigate the effect of Dunaliella salina, a microalga and its isolated zeaxanthin on age-related hepatic steatosis as well as their underling mechanism. Study Design. Age-related hepatic steatosis was induced in rats by intraperitoneal injection of D-galactose (200 mg/kg/day) for eight consecutive weeks. D. salina biomass (BDS; 450 mg/kg), its polar fraction (PDS; 30 mg/kg), carotenoid fraction (CDS; 30 mg/kg), and isolated zeaxanthin heneicosylate (ZH; 250 µg/kg) were orally administered to D-galactose treated rats for two weeks. METHODS: Blood samples were collected 24 hours after the last dose of D. salina treatments, animals were sacrificed, and liver tissues were isolated. Sera as well as hepatic tissue homogenates were used for further investigations. Liver tissues were also used for histopathological and immunohistochemical examinations. A computed virtual docking study for the biologically active candidates was performed to confirm the proposed mechanism of action. RESULTS: Oral treatment of D-galactose-injected rats with BDS, PDS, CDS, or ZH ameliorated the serum hepatic function parameters as well as serum levels of adiponectin, apolipoprotein B 100, and insulin. Furthermore, D. salina decreased the hepatic lipid contents, redox status biomarkers, inflammatory cytokine, and showing antiapoptotic properties. Molecular docking of ß-carotene and zeaxanthin on various receptors involved in the pathophysiological cascade of steatosis highlighted the possible mechanism underlying the observed therapeutic effect. CONCLUSION: D. salina carotenoids have beneficial effect on age-related hepatic steatosis in senescence rats through the regulation of redox status, inflammatory indices, and apoptotic biomarkers.

The ameliorating effect of carotenoid rich fraction extracted from Dunaliella salina microalga against inflammation- associated cardiac dysfunction in obese rats.

The carotenoid rich fraction of microalgae Dunaliella salina (crf-DS) have been receiving great attention, due to they abilities to protect and improve various disorders. The objective of this study is to explore the therapeutic efficiency of crf-DS on obesity-assciated cardiac dysfunction in the high-fat diet (HFD) treated rats. These rats were orally administered with crf-DS (150 mg /kg body weight), for six consecutive weeks in comparison with reference drug(orlistat). Specific cardiac biomarkers were examined including; adiponectin, plasminogen activator inhibitor (PAI-1), glucagon, troponin-I (cTnI). The cell adhesion molecules (VCAM and ICAM), C-reactive protein (CRP), collagen type II (Col II), collagen alpha-1 (III) chain (Col3A1), lipoxygenase activity (LOX), as well as histopathological examination of cardiac tissue were investigated. Results indicated a significant reduction(P ≤ 0.05) in adiponectin and glucagon levels in serum of obese rats. However, cTnI, PAI-1, cell adhesion molecules, CRP, Col II, and Col3A1 and LOX levels declared marked increase. Histopathological examination of cardiac tissue showed fibrosis with severe congestion in the myocardial blood vessels. On the other hand, rats medicated with a crf-DS demonstrated noticeable ameliorating effect in all the measured parameters. Beside, myocardial tissue of obese rats showed no alteration. Hence, It could be concluded that, oral supplementation with crf-DS is able to attenuate cardiac dysfunction in obese rats. Further extended work is needed to exploit, the possible application of D. salina as nutraceuticals and food additives.

Toxicity assessment of the green Dunaliella salina microalgae.

The chronic toxicity of the Dunaliella salina microalgae was examined to evaluate its toxicity by the exposure of laboratory animals to high doses of Dunaliella salina and to estimate the possibility of using it as a safe supplement. Different hematological and biochemical analysis including complete blood picture (CBC), liver function enzyme activities; aminotransferases (ALT and AST), alkaline phosphatase (ALP), total bilirubin, kidney function tests; urea, creatinine, and albumin, as well as blood glucose level, were measured. The histopathological investigation was also carried out on hepatic, renal and cardiac architectures to examine its safety. Treatment with the dose 100 mg /kg body weight of D. salina powder daily for three consecutive months did not show any signs of toxicity in both genders and in mice and rats (no mortality, no hair loss, no diarrhea, no patches of yellow color appearance, etc…..). Moreover, abnormalities on behavior, food and water intakes and health status among the treated animals were not observed. CBC profile revealed a significant increase in hemoglobin (Hb) level in treating male and female mice and rats compared to their related control levels. The biochemical analysis clearly showed an insignificant change in liver enzyme activities, blood glucose level at a dose of 100 mg/kg. Also, an insignificant reduction in total urea and creatinine levels in both genders of mice and rats were noticed. Histopathological investigation showed normal architectures of all organs. Hence we can conclude that Dunaliella salina has been proven a safe profile up to 100 mg/kg body weight, however, it succeeded to stimulate the Hb synthesis compared to control groups, showing its benifits to be used safely as food additives or protective and curative agent in different diseases in future.

Intestinal injury can be effectively prevented by Dunaliella salina in gamma irradiated rats.

Dunaliella salina (D. salina) is one of the most common microalgae that is used as human food. It is isolated from the salty lakes in El-Fayoum and Lake of Bardawil-Sinai in Egypt and can withstand very high concentrations of salt: The potentiality of D. salina, a unicellular biflagellate green alga to protect against intestinal injury induced after radiation exposure was studied. D. salina was given orally in doses of 100 and 200 mg/kg to male Wistar rats for 5 days before exposure to 6 Gray (Gy) gamma radiation and continued for a further two days. Rats were sacrificed 24 h later and intestinal segments were dissected out. One segment was examined histologically and another was used to prepare homogenates to assess relevant biochemical parameters reflecting intestinal injury. Radiation exposure led to a rise in the histological damage score, an increase in tissue tumor necrosis factor (TNF-α), interleukin (IL-1ß) and thiobarbituric acid reactive substances (TBARS) but a reduction in tissue reduced glutathione (GSH) and in serum citrulline. Pretreatment with either dose of D. salina effectively reduced the severity of intestinal mucositis induced by gamma radiation.

Heamatococcus pluvialis ameliorates bone loss in experimentally-induced osteoporosis in rats via the regulation of OPG/RANKL pathway.

BACKGROUNDS: Osteoporosis prevailing in elderly involves a marked increase in bone resorption showing an initial fall in bone mineral density leading to a significant reduction in bone formation. AIM: The present study aimed to investigate the effect of Heamatococcus pluvialis microalgae on osteoporosis in D-galactose-treated rats. The underlying mechanism was tracked targeting the osteoprotegerin (OPG)/ nuclear factor-κß ligand (RANKL) pathway using micro-computed tomography scanning. METHODS: Osteoporosis was induced in rats by intraperitoneal injection of D-galactose (200 mg/kg/day) for eight consecutive weeks. Osteoporotic rats were orally treated with H. pluvialis biomass (BHP; 450 mg/kg), its polar (PHP; 30 mg/kg) and carotenoid (CHP; 30 mg/kg) fractions for the last 2 weeks of D-Gal injection. Twenty four hours after the last dose of the treatments, tibia bones of the rats were scanned using micro-computed tomography scanning for bone mineral density (BMD), bone volume fraction (BV/TV), trabecular thickness/separation/number (Tb.Th, Tb.Sp, Tb.N) evaluation, blood samples were withdrawn and sera were used for biochemical assessment. Moreover, femur bones were examined histopathologically using several stains. RESULTS: Induction of osteoporosis was associated with a marked reduction in BMD, BV/TV, Tb.Th, Tb.Sp, Tb.N and in serum levels of phosphorus and catalase. On the other hand, a significant elevation in serum levels of calcium, bone alkaline phosphatase (BALP) and interleukin-6 was observed. Moreover, up-regulation of OPG was detected in osteoporotic rats. Oral treatment with BHP, and PHP incremented tibia BMD and serum phosphorus level along with the decrease in serum levels of calcium, BALP, interleukin-6, OPG and RANKL. However, treatment with CHP almost restored all the fore mentioned parameters to normal values. Furthermore, the histopathological evaluation emphasized the biochemical outcomes. CONCLUSION: H. pluvialis fractions rich in astaxanthin ameliorated bone loss in experimentally-induced osteoporosis in rats probably through the down-regulation of serum OPG in concurrence with up-regulation of serum RANKL.

Astaxanthin-Rich Haematococcus pluvialis Algal Hepatic Modulation in D-Galactose-Induced Aging in Rats: Role of Nrf2.

Purpose: Aging is associated with hepatic morphological and physiological deterioration due to the accumulation of endogenous and exogenous free radicals and the resultant oxidative stress. The present study aims to investigate the effect of Haematococcus pluvialis microalgae on hepatic changes associated with D-galactose (D-Gal)-induced aging in rats. Methods: Aging was induced in rats by daily intraperitoneal injection of D-Gal (200 mg/kg/day) for eight consecutive weeks. D-Gal-injected rats were treated by astaxanthin (ATX)-rich H. pluvialis biomass, its carotenoid and polar fractions for two weeks. Twenty four hours after the last dose, blood samples were collected and the liver tissues were isolated for further biochemical and histopathological examinations. Results: D-Gal induced aging was associated with an elevation in serum liver function parameters, hepatic oxidative stress biomarkers viz., catalase (CAT), glutathione transferase (GST) and myeloperoxidase (MPO), as well as decreased expression of nuclear factor like-2 (Nrf2). Moreover, induction of aging exhibited an elevation of hepatic inflammatory cytokine; interleukin-6 (IL-6) and its modulator; nuclear factor Kappa B (NF-KB). However, treatment of D-Gal injected rats with ATX-rich H. pluvialis restored the serum liver function parameters as well as hepatic CAT, GST and MPO levels with an elevated expression of Nrf2. Treatment with ATX-rich H. pluvialis was also accompanied with a decrease in hepatic levels of NF-KB and IL-6. Histopathological examination emphasized all the previous results. Similarly, all trans-astaxanthin showed high affinity towards Nrf2 with -7.93 kcal/mol estimated free energy of binding as well as moderate affinities towards IL-6 and NF-KB through a docking study. Conclusion: ATX-rich H. pluvialis showed beneficial effects by ameliorating the hepatic changes associated with D-Gal induced aging in rats due to its modulatory role of the Nrf2/Keap pathway.

Scenedesmus obliquus: Antioxidant and antiviral activity of proteins hydrolyzed by three enzymes.

PURPOSE: To obtain protein hydrolysates from fresh water green algae Scenedesmus obliquus by three different enzymes and evaluate its antioxidant and antiviral activity. METHODS: Enzymatic hydrolysates of green algae Scenedesmus obliquus protein were prepared by treatment with: 1.2% solution of pepsin, trypsin or papain. Protein was extracted from S. obliquus by three different extraction methods. Protein extracts and hydrolysates were assessed from stained gels following SDS-PAGE of samples. Antioxidant activity of protein hydrolysates was investigated. RESULTS: S. obliquus cells and protein extracts were rich in Arg, Lys, Asp, Ala, and His. Protein hydrolyzed by papain (Sd1pa) and protein hydrolyzed by trypsin (Sd2Try) induced highest antioxidant activity based on 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical-scavenging (41.41% and 40.62%) respectively, and on 2,2'-azinobis 3-ethyl-benzothiazoline-6-sulphonate (ABTS) radical (87.03% and 45.12%) respectively, at 150 µg/ml. The inhibitory effect and mode of action of protein hydrolysates were evaluated against Coxsackie B3 virus (CVB3). Protein hydrolyzed by papain (Sd2pa) and protein hydrolyzed by pepsin (Sd1pep) at 100 µg/ml exhibited antiviral activity (66.2% and 57.6%, respectively), against (CVB3) from all protein hydrolysates. CONCLUSION: S. obliquus protein hydrolysates have a potential as antioxidative neutraceutical ingredients and a potential therapeutic agent against CVB3.

Chemical Profile and Biological Activity of Casimiroa Edulis Non-Edible Fruit`s Parts.

Purpose: the non-edible fruit parts of Casimiroa edulis Llave et were evaluated for their active constituents and their potential as antioxidants, anti-inflammatory and antitumor activity. Methods: Fruits peel (FP) and seeds kernel (SK) of Casimiroa edulis Llave et Lex. were extracted successively with hexane and then methanol. Fatty acids were prepared from hexane extracts and identified by GC. Total flavonoid, phenolic acids and tannins contents in methanol extracts were determined by UV spectrophotometer and identified by HPLC. Antioxidant, in-vitro anti-inflammatory activity and antitumor effect against Caco-2 cell line were determined. Results: GC analysis of hexane extracts showed that oleic acid (47.00%) was the major unsaturated fatty acids in both extracts while lignoceric acid (15.49%) is the most abundant saturated fatty acid in (FP). Total phenolic, flavonoid and tannin contents in (FP) & (SK) methanol extracts were; 37.5±1.5, 10.79±0.66 and 22.28±0.23 for (FP); 53.5±1.5mg/g, 14.44±0.32 mg/g; and 53.73±3.58 mg/g for (SK) respectively. HPLC analysis of methanol extract revealed that; the major phenolic compound was pyrogallol in (FP) and p-hydroxybenzoic acid in (SK), the major flavonoid was luteolin 6-arabinose-8-glucose in (FP) and acacetin in (SK). Conclusion: This study showed that non-edible parts of C. edulis fruit is a rich source of different phenolic compounds and fatty acids which has great antioxidant, anti-inflammatory and antitumor activities; that could be used as a natural source in pharmaceutical industry.

Regional view of a Trans-African Drainage System.

Despite the arid to hyperarid climate of the Great Sahara of North Africa, pluvial climates dominated the region. Radar data shed some light on the postulated Trans-African Drainage System and its relationship to active and inactive tributaries of the Nile basin. Interpretations of recent elevation data confirm a source of the river water from the Red Sea highlands did not connect the Atlantic Ocean across Tushka basin, highlands of Uwinate and Darfur, and Chad basin, but northward to the ancestral Nile Delta. Elements of topography and climate were considered. They show that the former segments of the Nile closely mirror present-day tributaries of the Nile basin in drainage geometry, landscape, and climate. A rainfall data interpolation scenario revealed that this basin received concurrent runoff from both flanks such as Gabgaba-Allaqi to the east and Tushka basin to the west, similar to present-day Sobat and White Nile tributaries, respectively. Overall the western tributaries such as those of Tushka basin and Howar lead to the Nile, which was (and still is) the biggest river system in Africa.