Spondias which is widely used in traditional medicine for the treatment of many diseases. Spondias is a genus of flowering plants belonging to the cashew family (Anacardiaceae). This genus comprises 18 species distributed across tropical regions in the world. A variety of bioactive phytochemical constituents were isolated from different plants belonging to the genus Spondias. Diverse pharmacological activities were reported for the genus Spondias including cytotoxic, antioxidant, ulcer protective, hepatoprotective, anti-inflammatory, antiarthritic, and antidementia effects. These attributes indicate their potential to treat various degenerative diseases. The aim of this review is to draw attention to the unexplored potential of phytochemicals obtained from Spondias species, thereby contributing to the development of new therapeutic alternatives that may improve the health of people suffering from degenerative diseases and other health problems.It is believed that many degenerative diseases are due to oxidative stress. In view of the limited drugs available for treating degenerative diseases, natural products represent a promising therapeutic strategy in the search for new and effective candidates for treating degenerative diseases. This review focuses on the genus Degenerative disease results from a continuous process based on degenerative cell changes of tissues and organs, which increasingly deteriorate over time. This might happen due to normal bodily wear or lifestyle choices such as lack of exercise or eating habits. Oxidative stress is known to be implicated in the development of degenerative diseases. An imbalance between formation and neutralization of free radicals leads to oxidative stress. These reactive species seek stability through electron pairing with biological macromolecules such as proteins, lipids, and DNA in healthy cells leading to protein and DNA damage along with lipid peroxidation . These c Spondias belongs to family Anacardiaceae which comprises 70 genera and 600 species and is endogenous mostly in the tropics and subtropics worldwide but also extends into the temperate zone. Members of this family are used in traditional medicine in the treatment of many ailments -α-rhamnopyranosyl-(1→6)-β-glucopyranoside were isolated from the methanol extract of S. venulosa leaves [ S. purpurea fruit [ S. pinnata bark [Among the isolated phenolic compounds, geraniin and galloyl geraniin were isolated from the 80% ethanolic extract ofnd stems . Galloylpurpurea . Moreovea leaves . Gallic ea fruit . Furtherata bark .β-amyrin and oleanolic acid, were isolated from the methanolic extract of S. pinnata fruit [β-sitosterol, and β-sitosterol β-D-glucoside were isolated from the ethanolic extract of S. pinnata aerial parts [ S. mombin bark [ S. pinnata bark [ S. mombin and S. purpurea leaves [O-β-D-galactopyranosyl (1→5)-O-β-D-xylofuranoside were isolated from the ethanolic extract of S. mangifera roots [Triterpenoid compounds, including ta fruit . Sterolsal parts , while sbin bark . Ergosteata bark . In addia leaves . Some sara roots . Spondias species. Hydrodistillation of the leaves of S. mombin and S. purpurea led to the isolation and identification of α-pinene, β-pinene, caryophyllene, humulene, indene, and cadinene [Various volatile oil constituents were reported from differentcadinene . S. pinnata showed nutritional value and was found to be rich in several amino acids, namely, glycine, cysteine, serine, alanine, and leucine [O-methylglucose, 2,3,6-tri-O-methylglucose, 2,3-di-O-methylglucose, and 3-O-methylglucose were isolated from the aqueous extract of S. pinnata fruit [β-D-glucopyranosyl-(6′→1′′)-β-D-glucofuranoside (an acid glycoside) was obtained from the ethanolic extract of S. pinnata fruits [ Spondias are presented in Figures The fruit of leucine . Moreoveta fruit . Propan-a fruits . The chenosyl-6′→′′-β-D-gl Spondias are detailed below.Different reported pharmacological activities of the genus S. pinnata bark exhibited significant cytotoxicity on human lung adenocarcinoma (A549) and human breast adenocarcinoma (MCF-7) cell lines via inducing apoptosis. In vitro WST-1 cell proliferation assay was carried out; A549 cells were seeded in a 96-well culture plate at a density of 5∗104 cells/well whereas MCF-7 cells were seeded at 1∗104 cells/well and allowed to settle for 2 h. The cells were then treated with the methanolic extract of S. pinnata ranging from 0 to 200 μg/ml for 48 h. The 70% methanolic extract of S. pinnata inhibited the growth of both A549 and MCF-7 cells in a dose-dependent manner with an IC50 value of 147.84 and 149.34 μg/ml, respectively. Cell proliferation and viability were quantified by measuring the absorbance of the produced formazan at 460 nm using a microplate ELISA reader. The pathway of apoptosis induction may be due to an increase in Bax/Bcl-2 ratio in both cell types, which resulted in the activation of the caspase cascade, subsequently leading to cleavage of poly adeno ribose polymerase enzyme [Ghate et al. (2013) demonstrated that the methanolic extract ofe enzyme . S. pinnata for their cytotoxic activity against human glioblastoma cell line (U87). In vitro WST-1 cytotoxicity assay was carried out; 1 × 104 cells were treated with compounds isolated from the ethyl acetate fraction (1 to 30 μg/ml) for 48 h in a 96-well culture plate. Two isolated compounds  showed promising cytotoxic activities with IC50 of 59.28 and 8.44 μg/ml, respectively [ Moutan Cortex Radicis enhances the antitumor effects through modulation of the function of CD4+CD25+ Treg cells. In vitro, methyl gallate decreased CD4+CD25+ Treg cell migration and reduced the suppressive function of effector T-cells. In tumor-bearing animals, treatment with methyl gallate delayed tumor progression and prolonged survival through inhibition of the tumor infiltration of CD4+CD25+ Treg cells [Chaudhuri et al. (2015) tested the activity of compounds isolated from the ethyl acetate fraction obtained from the bark ofectively . Gallic ectively . Previoueg cells . S. mangifera bark is a potent source of antioxidants. Total antioxidant activity was assessed in vitro, depending on the ability of the 70% methanolic extract to scavenge ABTS radical cation, and compared to trolox standard, the total antioxidant activity of the 70% methanolic extract was calculated from the decolorization of ABTS cation which was measured spectrophotometrically at 734 nm; the trolox equivalent antioxidant value was of 0.78 [ S. mangifera methanolic fruit extract at concentration of 5 μg/ml showed 16% radical scavenging activity compared to the same concentration of vitamin C which showed only 5% radical scavenging activity [Hazra et al. (2008) proved that the 70% methanolic extract of of 0.78 . In addiactivity . S. mangifera fruits contains large amounts of phenolics, flavonoids, and acid glycosides, such as propan-1,2-dioicacid-3-carboxyl-β-D-glucopyranosyl-(6′→1′′)-β-D-glucofuranoside. In vitro and in vivo studies were conducted to test the effects of ethanolic extract and acid glycoside as antioxidants against anoxia-stress tolerance, swimming endurance, and cyclophosphamide—immune suppression. The antioxidant activity was compared to a standard drug Geriforte [Arif et al. (2016) showed that the ethanolic extract oferiforte .An in vitro study was carried against DPPH∙ and determined by a UV spectrophotometer at 517 nm. Aliquots of 0.05, 0.5, and 1 mg/ml of either the ethanolic extract of the acid glycoside were mixed in test tubes each containing 3 ml of methanol and 0.5 ml of 1 mM DPPH∙; ascorbic acid was used as a standard at the same concentrations, and the reaction mixture was incubated at 37°C for 30 min. The radical scavenging activity was calculated; IC50 was 0.32 and 0.15 mg/ml for the ethanolic extract and acid glycoside, respectively, while IC50 of ascorbic acid was 0.015 mg/ml. These results indicated that the ethanolic extract and the acid glycoside exhibited a significant antioxidant activity [activity . in vivo experiment was carried out on thirty Swiss Albino mice which were divided into five groups of six mice each; group 1 served as the control and received vehicle alone (2% gum acacia), groups 2 and 3 were treated with 100 and 200 mg/kg/day of the ethanolic extract, group 4 was treated with 10 mg/kg/day of the acid glycoside, and group 5 was treated with 50 mg/kg/day of the standard drug Geriforte; all the groups were treated for 3 weeks; every week after 1 h of drug administration, each animal was placed in an airtight glass container of 250 ml and the time taken for appearance of generalized clonic seizures was observed . Thereafter, the mice were removed for recovery; the time duration from the entry of the animal into the hermetic vessel to the appearance of the first convulsion was taken as the time of anoxia tolerance; the anoxia tolerance effect was increased with increasing dose and duration of treatment, indicating the significant stress relaxant activity [Furthermore, anactivity . in vivo study was carried out on 30 mice, divided into five groups of six mice each; group 1 served as the control and received vehicle alone (2% gum acacia), groups 2 and 3 were treated with 100 and 200 mg/kg/day of ethanolic extract, respectively, group 4 was treated with 10 mg/kg/day of acid glycoside, and group 5 was treated with 50 mg/kg/day of the standard drug Geriforte; all the drugs were given orally once a day for seven days; on the seventh day, 1 hour after drug administration, all the mice were made to swim in a water tank maintained at room temperature until they sank; the control group swam for 131.2 min; the ethanolic extract treated mice at doses of 100 and 200 mg/kg/day swam for 152.7 and 158.6 min, respectively, whereas the acid glycoside treated mice swam for 155.4 min. It was evident that the ethanolic extract and the acid glycoside treated mice exhibited a significant increase in physical swimming endurance time [Anothernce time . in vivo study was carried out on 24 mice, divided into four groups of six mice each. It was observed that the administration of cyclophosphamide alone (25 mg/kg/day) produced a significant decrease in the total RBCs and leukocytes counts, whereas cyclophosphamide given along with ethanolic extract (100 mg/kg/day) and acid glycoside (10 mg/kg) conferred a good protection by increasing the haematological parameters. It was suggested, based on this study, that the ethanolic extract and acid glycoside may be coadministered with chemotherapy for the treatment of patients with severely impaired or suppressed immune system [An extrae system , as the e system . in vivo study on Wistar rats to show the effects of combining conventional chemotherapy with S. pinnata bark extract to reduce chemotherapy's side effects. The rats were divided into four groups: group 1 , group 2  in a single dose of 60 mg/kg b.w.), group 3 (received etoposide followed by S. pinnata bark extract (100 mg/kg b.w.) orally once a day from 0 h to 72 h), and group 4 (received etoposide (i.p.) followed by S. pinnata bark extract in a dose of 200 mg/kg b.w. orally once a day from 0 h to 72 h). The results showed that animals which received chemotherapy in group 2 showed a significant decrease of GSH level in the liver and kidney tissues as compared to the control group, while treatment with S. pinnata bark extract after chemotherapy showed a significant increase in GSH level when compared to group 2. This study proved the protective action of the extract on the liver and kidney against chemotherapy-induced chemical stress [Shetty et al. (2016) conducted anl stress . S. mombin leaves showed a significant antioxidant activity, and in an in vitro DPPH∙ assay, the hydroethanolic extract was tested at 60, 125, 250, and 500 μg/ml and showed DPPH∙ radical scavenging activity ranging from 66% to 76% [Cabral et al. (2016) proved that the hydroethanolic extract of% to 76% . S. purpurea fruit showed a strong free radical scavenging activity and this was deduced by carrying out an in vitro study to evaluate the ability of the methanol extract of S. purpurea fruit to sequestrate the DPPH∙ radicals; the flavonoid rutin was used as a positive control and sequestrated 90.01% of the DPPH∙ radicals at concentration 250 μg/ml. while the methanol extract of S. purpurea fruit sequestrated 74.41% with EC50 of 27.11 μg/ml [ Spondias has been attributed mainly to their flavonoids and phenolic content [The methanol extract of11 μg/ml . The str in vivo study to evaluate the ulcer protective activity of S. mangifera methanolic bark extract. Gastric ulceration was achieved by administering different doses of indomethacin  to rats orally and 100 mg/kg was found to be the most effective for producing gastric ulceration in the rats. The rats were then divided into four groups, each comprising six animals. Food and water were withdrawn 24 h and 2 h, respectively, before drug administration. Rats in group 1 received 100 mg/kg indomethacin while those in group 2 were pretreated with 100 mg/kg cimetidine. The rats in groups 3 and 4 were pretreated with 100–200 mg/kg of bark extract 1 h prior to the administration of indomethacin (100 mg/kg). The drugs were administered intragastrically. After 4 h, the animals were killed by cervical dislocation and their stomachs were removed and opened along the greater curvature. The ulcer index (UI) of each group was calculated. The groups treated with bark extract showed a marked reduction of the ulcerogenic effect of indomethacin, reducing the ulcer index from 17.7 (ulcerated control) to 8.7 and 6.7 for the groups treated with bark extracts of 100 mg/kg and 200 mg/kg, respectively. The methanolic bark extract of S. mangifera was thus concluded to possess a marked inhibitory effect of indomethacin-induced ulceration [The pathophysiology of gastric ulceration involves an imbalance between offensive and protective factors , 45. Ariceration . S. mombin leaves. Ulceration was induced in Albino rats by oral administration of indomethacin which caused a significant increase in the degree of ulceration. Pretreatment with the extract of 200 mg/kg b.w. facilitated the ulcer healing process, which was associated with a decrease in pepsin activity and an elevation in mucin levels in the gastric mucosa. Moreover, S. mombin leaf extract ameliorated the oxidative stress and inhibitory action of indomethacin on prostaglandin synthesis [Sabiu et al. (2015) tested the gastroprotective and antioxidative potential of the aqueous extract ofynthesis . S. pinnata stem heartwood possess a marked in vivo hepatoprotective effect on CCl4 intoxicated rats. The ethyl acetate and methanolic extracts were administered at doses of 100, 200, and 400 mg/kg, p.o., and the results showed a protective activity in a dose-dependent manner as evidenced by the significant decreases in ALT and AST to their normal levels, which was comparable to silymarin. The hepatoprotective effect in this study was attributed to the presence of flavonoids. Histopathological examination was also carried out on CCl4 intoxicated rats and revealed that normal hepatic architecture was retained in rats treated with S. pinnata extracts [The ethyl acetate and methanolic extracts ofextracts . S. pinnata stem bark methanol extract on iron-induced liver injury in mice. Intraperitoneal administration of iron dextran induced an iron overload and led to liver damage along with a significant increase in serum hepatic markers . The administration of S. pinnata methanol extract in doses of 50, 100, and 200 mg/kg induced a marked increase in antioxidant enzymes, along with dose-dependent inhibition of lipid peroxidation, protein oxidation, and liver fibrosis. Meanwhile, the levels of serum enzyme markers and ferritin were also reduced, suggesting that the extract is potentially useful as an iron chelating agent for iron overload diseases [Hazra et al. (2013) evaluated the effect ofdiseases . S. pinnata bark against iron-induced liver fibrosis and hepatocellular damage. In an iron-overloaded liver, iron reacts with cellular hydrogen peroxide to generate hydroxyl radicals which in turn initiate the propagation of various free radicals; this situation leads to oxidative stress. Two compounds  were isolated from the ethyl acetate fraction of this extract; an in vivo study showed that methyl gallate exhibited better iron chelation properties than gallic acid. It was proved that methyl gallate overcomes hepatic fibrosis by ameliorating oxidative stress and sequestrating the stored iron in cells [ in vivo protective effect of gallic acid isolated from Peltiphyllum peltatum against sodium fluoride induced hepatotoxicity and oxidative stress. The results showed that gallic acid (10 and 20 mg/kg) prevented the sodium fluoride induced abnormalities in the hepatic biochemical markers; these effects were comparable to the reference drug silymarin (10 mg/kg) [Chaudhuri et al. (2016) evaluated the activity of the methanolic extract ofin cells . These r0 mg/kg) . S. purpurea fruit was assessed in vitro by the trans-resveratrol method, which indicated its marked photoprotective ability against UVA radiation [ in vitro UVB photoprotection effect of the ethanol extract of S. purpurea fruit by a spectrophotometric method. The photoprotective effect was attributed to phenolic compounds in S. purpurea fruit extract having the ability to absorb the solar radiation, to scavenge free radicals, and to decrease the harmful effects of the sun [Ultraviolet A and ultraviolet B are known to induce skin cancer. The free radicals generated from sunlight are responsible for the degradation of essential cellular components such as DNA and proteins . The UVAadiation . Silva e the sun . S. mombin leaves showed a significant anti-inflammatory activity in a carrageenan-induced peritonitis model in mice. Carrageenan induced neutrophil migration to the peritoneal cavity and typical signs of acute inflammation including vasodilation, edema, and leukocyte infiltration. It was evident from this study that S. mombin leaf extract  reduced the leukocyte influx to the peritoneal cavity of the treated animals [The hydroethanolic extract of animals . S. tuberosa leaves hydroethanolic extract. Furthermore, an in vivo study was conducted on Swiss Albino mice, where dexamethasone was used as a standard anti-inflammatory drug and carrageenan was used to induce hind paw edema. The extract  induced significant amelioration of the inflammatory response induced by carrageenan, a marked reduction in the number of leukocytes in the peritoneal cavity, and a significant decrease in myeloperoxidase activity [da Silva Siqueira et al. (2016) showed that phenolic compounds were responsible for the anti-inflammatory activity exhibited byactivity . S. pinnata inhibits nitrite formation in vitro by directly competing with oxygen in the reaction with nitric oxide. The results revealed that IC50 of the methanolic extract (tested at 200 μg/ml) was 716.32 μg/ml which was lower than that of the reference compound gallic acid (IC50 = 876.24 μg/ml). The scavenging percentages were 22.3 and 15.8% for S. pinnata and gallic acid, respectively. This study proved that the extract exhibited more potent peroxynitrite radical scavenging activity than the standard gallic acid [Nitric oxide plays an important role in various inflammatory processes. However, sustained levels of production of this radical are directly toxic to tissues and contribute to the vascular collapse associated with septic shock, whereas chronic expression of nitric oxide radical is associated with various degenerative diseases, including carcinomas and inflammatory conditions such as juvenile diabetes, multiple sclerosis, arthritis, and ulcerative colitis. The toxicity of NO increases greatly when it reacts with a superoxide radical, forming the highly reactive peroxynitrite anion (ONOO-). Hazra et al. (2008) proved that the methanolic extract of S. mombin leaves  enhanced the learning and memory capabilities of Wister rats due to structural changes observed in the cerebrum. Improved learning and memory have been also linked to structural changes of the limbic system [ in vivo protective effect of the hydroethanolic leaf extract of S. mombin  and proved the protective effect against scopolamine-induced cognitive dysfunction and memory deficit that could be attributed to the extract antioxidant properties [The ability to acquire knowledge and to retain this acquired knowledge can be defined as learning and memory. Several conditions such as aging and stress may lead to the impairment of learning. It has been shown that aging may lead to various neurodegenerative processes including memory loss, dementia, and Alzheimer's disease . Asuquo c system . The aquc system , 57. Ishoperties . S. pinnata bark. The analgesic activity was evaluated using acetic acid, formalin test, and hot plate model. The extract showed a dose-dependent analgesic effect  in the acetic acid test, comparable to the effect of acetyl salicylic acid. Terpenoids, flavonoids, and tannins were responsible for the analgesic activity [ S. pinnata bark ethanol extract . Pyrexia was induced in Albino rats by brewer's yeast. The extract showed a significant reduction in pyrexia, which continued for 5 hours after drug administration [Panda et al. (2009) tested the analgesic activity of the ethanolic extract ofactivity . Panda estration . S. pinnata fruit at the concentration of 10 mg/ml have a significant thrombolytic activity compared to streptokinase as a standard substance [ S. pinnata (1 mg/ml) leaves has a membrane stabilizing activity for human RBCs in hypotonic solution-induced hemolysis. In case of heat-induced hemolysis, S. pinnata extracts produced marked inhibition of hemolysis [ S. pinnata aerial parts and its different fractions. The ethyl acetate fraction exerted the highest thrombolytic activity and membrane stabilizing activity [Manik et al. (2013) showed that both ethyl acetate and aqueous extracts ofubstance . Kamal eemolysis . Uddin eactivity . Spondias. The leaves of S. mombin were tested in vitro by Fred-Jaiyesimi et al. (2009) for their hypoglycemic activity. A new compound, 3β-olean-12-en-3-yl (9Z)-hexadec-9-enoate, isolated from the diethyl ether fraction of the methanolic extract of S. mombin leaves, showed an α-amylase inhibitory activity similar to the activity of acarbose. The methanolic leaf extract and the isolated new compound decreased postprandial hyperglycemia. The methanolic extract (250 mg/ml) showed 39% inhibition of the α-amylase activity, while the diethyl ether fraction (70 mg/ml) showed 73% inhibition and the isolated compound (20 mg/ml) exhibited 57% α-amylase inhibition [The hypoglycemic activity was tested using different extracts of the genusi et al. 009 for t S. pinnata, which was comparable to glibenclamide. The test was carried out in vivo, and the methanolic extract was administered at a dose of 300 mg/kg to rats. After 30 min of treatment, rats were loaded orally with glucose . Blood samples were collected before and at 30, 90, and 150 min intervals after glucose administration, the methanol extract was found to reduce blood glucose level by 63.12%, and the results were found to be comparable to glibenclamide [Mondal and Dash (2009) showed a promising hypoglycemic effect of the methanolic bark extract ofnclamide . S. pinnata roots in vivo using oral glucose tolerance test and indicated a significant decrease in blood glucose levels after four hours of treatment as compared to glibenclamide [Acharyya et al. (2010) tested the hypoglycemic activity of both the methanolic and the aqueous extracts ofnclamide . S. mombin leaves on anterior pituitary, ovary, uterus, and serum sex hormones. The animals received the ethanolic extract at dose levels of 250, 350, and 500 mg/kg b.w. The results showed a significant decrease in the weight of pituitary, ovary, and uterus of the treated animals, along with a significant reduction in FSH, LH, estradiol, and progesterone levels. Therefore, this study concluded that the extract showed antifertility activity and can be used as a contraceptive [Asuquo et al. (2013) carried out a study on adult female Wister rats to determine the effect of the ethanolic extract ofaceptive . in vitro antihypertensive activity of the aqueous extract of S. pinnata fruit (20 μg/ml). The angiotensin-converting-enzyme inhibitory activity was assayed using ACE from rabbit lung and N-hippuryl-L-histidyl-L-leucine as a substrate. This showed 50% inhibition of ACE enzyme [Das and De (2013) tested theE enzyme . in vitro antibacterial activity of the methanolic and the aqueous extracts of S. pinnata bark by cup plate diffusion method at the concentrations of 50, 100, and 150 mg. The activity was tested against Escherichia coli, Salmonella Typhimurium, and Vibrio cholerae and compared with penicillin and streptomycin as standard drugs. The methanolic extract showed a good antibacterial activity against Gram +ve and Gram −ve bacteria, while the aqueous extract showed only a mild antibacterial activity. The resin of S. pinnata also showed an antibacterial activity against Bacillus subtilis [Arif et al. (2008) tested thesubtilis . S. pinnata fruits showed a strong antibacterial activity against both Gram +ve and Gram −ve bacteria. The antimicrobial activity was tested by disc diffusion method; standard discs of kanamycin (30 μg/disc) and blank discs were used as positive and negative controls, respectively [The 80% ethanolic extract ofectively . S. pinnata bark and tested their antipseudomonal activity by agar disc diffusion method against a moderately resistant strain of Pseudomonas aeruginosa MTCC 8158. The tested organism was completely resistant to ampicillin and tetracycline at concentrations of 10 and 30 μg/disc, respectively, while exhibiting an inhibition zone of 15 mm against streptomycin at 100 μg/disc concentration. SP-40 exhibited an inhibition zone of 20 mm, which was better than streptomycin at comparable concentrations. SP-60, however, did not show any antimicrobial activity against this organism up to a concentration of 200 μg/disc. The MIC values of SP-40, thus, were estimated to be between 25 and 12.5 μg/disc [Tapan et al. (2014) isolated two new ergosteryl triterpenes  from μg/disc . S. mombin stem bark. Both compounds exhibited a marked antimycobacterial activity with 93% inhibition against Mycobacterium tuberculosis by a fluorometric microplate Alamar Blue Assay [Olugbuyiro et al. (2013) isolated two new phytosterols: stigmasta-9-en-3,6,7-triol and 3-hydroxy-22-epoxystigmastane from the methanolic extract ofue Assay . S. purpurea showed a strong antimicrobial activity against E. coli and P. aeruginosa using the disc diffusion method [ S. dulcis fruit [Furthermore, the methanolic fruit extract ofn method . Islam eis fruit . S. pinnata bark were tested for anthelmintic activity. Florido and Cortiguerra (2003) and Kumar et al. (2012) proved that the ethanolic extract with a concentration range of 50 mg/ml and 100 mg/ml showed more potent activity than the acetone extract [ S. pinnata was shown to exhibit an anthelmintic activity against Indian earthworms due to different glycosides present in the bark. Mondal et al. (2010) tested the anthelmintic activity of the chloroform extract of the bark of S. pinnata  and showed promising effects [The ethanolic and acetone extracts of extract , 71. The effects . S. pinnata bark (300 mg/kg) to Wister Albino rats produced significant diuretic and laxative activities as compared to reference standards furosemide and agar [Mondal et al. (2009) showed that the administration of chloroform and the methanol extracts ofand agar . in vivo study using the methanolic and ethanolic extracts of S. mombin leaves and showed promising antiepileptic and antipsychotic effects. They also tested the effects of aqueous, methanolic, and ethanolic extracts of S. mombin on hexobarbital-induced sleep in mice. Animals given hexobarbitone (100 mg/kg i.p.) showed loss of writhing reflex within five minutes of administration. The administration of the aqueous extract (100 mg/kg) decreased the latency of sleep significantly and was more potent in increasing hexobarbitone-induced sleeping time in mice. The methanolic extract did not alter the latency of sleep, whereas it increased the latency time at doses of 12.5 and 50 mg/kg. The three extracts produced a dose-dependent prolongation of hexobarbitone-induced sleeping time in mice [Ayoka et al. (2006) conducted an in mice . S. mombin leaves (≤5 g/kg) did not produce any toxic effects in mice and rats. Intraperitoneal administration of the aqueous extract (≤200 mg/kg) also did not produce any toxic effects; however, the ethanolic and methanolic extracts (>100 mg/kg) produced toxic symptoms. Lethal effects were observed in mice and rats with the three extracts at the dose of 3.2 g/kg administered i.p. LD50 in mice for ethanolic extracts was 480 mg/kg while it was 1.1 g/kg for the methanol extract and 1.36 g/kg for the aqueous extract. Also, LD50 in rats for the ethanolic, methanolic, and aqueous extracts was 620 mg/kg, 1.08, and 1.42 g/kg, respectively. The LD50 determination of the extracts was carried out in a 48 h continuous observation [It was evident that oral administration of aqueous, methanolic, and ethanolic extracts ofervation . in vivo toxicity of chloroform, methanol, and aqueous extracts of S. pinnata bark. The animals were divided into different groups of six animals each. The control group received 1% Tween-80 in normal saline . The other groups received 100, 200, 300, 600, 800, 1000, 2000, and 3000 mg/kg of the tested extracts, respectively, in a similar manner. Immediately after dosing, the animals were observed continuously for the first 4 h for any behavioural changes. They were then kept under observation for up to 14 days after drug administration to find out the mortality rate if any. It was found that the chloroform and methanol extract induced sedation, diuresis, and purgation at all tested doses. However, there was no mortality in any of the extracts at the tested doses till the end of the observation period [Mondal and Dash (2009) tested the acuten period . in vitro and in vivo models which could validate its ethnopharmacological use. This evidence of use and the absence of toxicity can provide an important basis for the development of herbal medicines from the aqueous extract of different Spondias species.Based on these results, it can be concluded that the aqueous extract is the safest one among the tested extracts. Furthermore, the aqueous extract showed a variety of pharmacological activities using different Spondias were widely used in traditional medicine due to their beneficial therapeutic effects. This is attributed to their diverse bioactive phytoconstituents like phenolics and flavonoids which possess marked antioxidant activity and thus are capable of preventing many degenerative diseases. The present review provides a comprehensive understanding of the chemistry and pharmacology of Spondias species, which may help in the discovery of new candidates for the treatment of various degenerative diseases and health problems.Presently, there is an increased demand worldwide for the use of natural remedies. Herbal medicines could be used as a complementary or alternative medicine to synthetic drugs, and this requires more laboratory investigations on their pharmacological activities. Many degenerative diseases are associated with oxidative stress. There is an increased demand worldwide for nontoxic, easily accessible, and affordable antioxidants of natural origin. Plants belonging to the genus


Hypertriglyceridemia is one of the three most common causes of AP, which is associated with the AP prognosis that has not been clearly defined.In this retrospective study, 1539 AP patients, who had serum triglyceride (TG) levels measured within the first 72 h, were assessed. The study groups consisted of patients with normal, mild, moderate, and severe/very severe HTG levels based on the Endocrine Society Clinical Practice Guidelines. We collected baseline demographic information, laboratory values, complications, and clinical outcome data in different HTG severity groups to analyze the clinical significance of elevated TG levels in AP.Ptrend < 0.001). acute necrotic collection (ANC) and pancreatic necrosis developed in 32 and 39 of 112 patients (29% and 35%)  in the severe/very severe HTG group, respectively. The proportion of persistent organ failure (POF), multiple organ failure (MOF), and persistent Systemic Inflammatory Response Syndrome (SIRS) increased with higher grades of HTG . The ICU admission rate was higher in the severe/very severe HTG group . A logistic multivariate regression analysis showed a positive correlation between HTG and certain AP complications.Our study included 1539 AP patients; of these, 1078 (70%) had a normal TG levels, and 461 (30%) had elevated TG levels. The rates of severe AP increased in HTG groups of increasing severity (4% vs. 8% vs. 12%; In addition to other factors, an elevated TG level could be associated with the severity and prognosis of AP, including pancreatic necrosis, POF, MOF, persistent SIRS, ICU admission, and mortality. Acute pancreatitis (AP) is a complex inflammatory disease that locally involves the pancreas as well as systemic organs. The AP incidence is approximately 70 cases per 100,000 individuals worldwide [Hypertriglyceridemia (HTG), which is one of the three most common causes of AP, occurs in 14.3% of AP patients . CompareThis is a retrospective single-center observational study. The study period was 8 years, occurring between January 1, 2005 and December 31, 2013. The study was approved by the ethics committee of The First Affiliated Hospital of Nanchang University (No. 2011001).The data were collected from an electronic medical database. Patients who were admitted to our hospital with an AP diagnosis and had serum TG levels measured within 72 h of presentation were included in the study. The selection process for patients is shown in a flow chart Fig. . AP was The diagnosis and severity classification of HTG were based on the Endocrine Society Clinical Practice Guidelines . An HTG Baseline demographic data, including age, gender, medical history, admission number, and date, were collected for all patients in the study. We collected data on the patients’ vital signs at admission, hemogram, blood biochemical parameters, and blood gas analysis.2 and Fisher’s exact tests. Logistic multivariate regression analyses were used for risk factors with categorical variables. A value of P < 0.05 was considered statistically significant.Statistical analyses were performed using IBM SPSS software, version 20.0 . Numerical data are expressed as the means ± SD, and categorical variables are expressed as the means (ratios). For continuous variable comparisons, an independent sample Student’s t test was used for two groups, and Cuzick’s trend test was used for multiple groups. Categorical variables were evaluated using the χP < 0.001) and younger  compared to patients with normal TG levels. A higher proportion of acute biliary pancreatitis was observed in the normal TG group, and more cases of alcoholic and HTG pancreatitis were found in the HTG group. Histories of diabetes mellitus , smoking , and alcoholism  were more common in the HTG group. No significant difference was identified with regard to hypertension . The baseline pancreatitis characteristics and etiologies in both groups are shown in Table The study included 1539 AP patients; of these, 1078 (70%) had normal TG levels (0.80 ± 0.37 mmol/L), and 461 (30%) had elevated TG levels (8.42 ± 8.91 mmol/L). The 461 patients with HTG were more commonly male , male (Ptrend < 0.001), have diabetes mellitus (Ptrend < 0.001), be alcoholic (Ptrend < 0.001), and smoke (Ptrend < 0.001), as presented in Table The stratified analysis of the HTG group showed that patients with greater HTG severity were more likely to be younger (Ptrend < 0.001). Other parameters recorded are presented in Table .Laboratory parameters on admission were compared among patients with different HTG severity categories. Some parameters, such as HCT, lactate dehydrogenase (LDH), total cholesterol (TC), and blood glucose (GLU), increased with higher grades of HTG (The 1539 AP patients included 327 (21%) acute peripancreatic fluid collection (APFC) cases, 242 (16%) acute necrotic collection (ANC) cases, 336 (22%) pancreatic necrosis cases, 345 (22%) persistent organ failure (POF) cases, 101 (7%) multiple organ failure (MOF) cases, 373 (24%) persistent SIRS cases, 438 (28%) ICU admissions, 87 patients (6%) who succumbed to their illness, and 37 (2%) high volume hemofiltration (HVHF) cases. The median hospital length of stay was 9 days.Ptrend < 0.001). ANC and pancreatic necrosis developed in 32 and 39 of 112 patients (29% and 35%)  with severe/very severe HTG, respectively. The proportion of POF, MOF and persistent SIRS increased with higher grades of HTG . The need for ICU admission was higher in the severe/very severe HTG group, accounting for 57/112 patients . One patient with a normal TG level required HVHF (0.1%), and 29 patients with severe/very severe HTG required HVHF (26%) (Ptrend < 0.001). No significant difference was noted in the development of APFC, length of stay, and mortality among the groups as shown in Table The rates of severe AP increased as the HTG severity classification of patients increased . A higher proportion of AP patients succumbed to their illness in the SAP group than in the MSAP and MAP groups . Similarly, 25 cases (7%) of HVHF were observed in the SAP group, which was at a rate higher than the rates observed in the MSAP (2%) and MAP (0.2%) groups (Ptrend < 0.001) as shown in Table AP patients with HTG pancreatitis had a higher proportion of MSAP (44%) and SAP (34%). TG and GLU levels increased as the AP severity increased . Early reports have indicated the respiratory insufficiency that is seen in AP could be mediated by TG elevations affecting pulmonary gas exchange and mechanics [SIRS that persists for 48 h or more after symptom onset is indicative of a poor prognosis . In our echanics . In the echanics , 18, 19.Recently, several studies have reported an effect of HTG on the clinical course of AP. Charlesworth et al. reported that HTG-induced AP was present in 2.3% of patients presenting with AP . PederseThe determinant of the severity of AP during the early phase is primarily the presence and duration of organ failure . The revIn summary, a more severe form of pancreatitis was observed in the higher TG groups than that observed in the lower TG groups, which implies that an elevated TG level may be associated with a poor prognosis, as indicated by a discrete proportional trend in our AP patients.


To provide a comprehensive quantitative assessment of nutritional status, digestion and absorption, and quality of life (QoL) in patients with locally advanced pancreatic cancer (LAPC).  Sixteen patients with LAPC were prospectively assessed for weight loss (WL), body mass index (BMI), fat-free mass index (FFMI), handgrip strength (HGS), dietary macronutrient intake, serum vitamin levels, resting and total energy expenditure , intestinal absorption capacity and fecal losses , exocrine pancreatic function ), and gastrointestinal quality of life (GIQLI). P = 0.002) than predicted REE. TEE was significantly higher than daily energy intake (P = 0.047). Malabsorption (<85%) of energy, fat, protein, and carbohydrates was observed in, respectively, 9, 8, 12, and 10 patients. FE1 levels were low (<200 μg/g) in 13 patients. Total QoL scored 71% (ample satisfactory).  Two patients had a low BMI, 10 patients had WL > 10%/6 months, 8 patients had a FFMI < P10, and 8 patients had a HGS < P10. Measured REE was 33% higher (NCT01939665. Patients with LAPC have a severely impaired nutritional status, most likely as a result of an increased REE and malabsorption due to exocrine pancreatic insufficiency. The trial is registered with PANFIRE clinicaltrials.gov  Pancreatic cancer is the fourth most common cause of cancer death in Western countries with an overall 2-year survival rate of less than 10% . CurrentIt is known that pancreatic cancer may affect the nutritional status through anorexia, an elevated resting energy expenditure (REE) as well as through exocrine pancreatic insufficiency (EPI), leading to fecal losses of energy, macro-, and micronutrients –11. SubsAlthough pancreatic cancer has been associated with a poor nutritional status and QoL, little quantitative data of these aspects is available in patients with LAPC , 10, 13.Sixteen outpatients with LAPC were prospectively included in the period between April 2014 and January 2015 at the interventional radiology ward of the VU University Medical Center (VUmc), Amsterdam, the Netherlands. A registered dietitian (JW) performed a comprehensive assessment of nutritional status (NS), digestion and absorption, and quality of life (QoL) within two weeks before experimental treatment with irreversible electroporation (IRE) of the pancreas [The Medical Ethics Review Committee of the VUmc approved this study, and included patients gave written informed consent for participation before entering the study (PANFIRE—pilot study: irreversible electroporation (IRE) to treat locally advanced pancreatic carcinoma, registered at clinicaltrials.gov NCT01939665).Data of patient characteristics were obtained prospectively from medical charts, defined by sex, age at diagnosis, tumor location, previous tumor treatment, medication, use of pancreatic enzyme replacement therapy (PERT), and previous treatment.The following parameters of nutritional status were determined: height, weight, body mass index, history of weight loss, body composition, handgrip strength, energy expenditure, and dietary energy and protein intake. Serum fat-soluble vitamin status was obtained from fasting blood samples.2) was calculated from weight in kilograms divided by height in meters squared. Weight loss history was inquired. Body composition was measured using bioelectrical impedance analyses using Bodystat© 1500MDD (EuroMedix) in patients without edema or ascites, in a supine position, after toilet visiting, and in a fasted state. Fat-free mass index (FFMI in kg/m2) was derived from fat-free mass in kilograms divided by height in squared meters.Height (cm) and weight (kg) were measured and performed with light clothing and without shoes. Body mass index  and at least one of either reduced BMI (<20 or <22 kg/m2 in subjects younger and older than 70 years) or a low FFMI  [The diagnostic criteria for malnutrition of The European Society of Clinical Nutrition and Metabolism (ESPEN) were used to define a poor nutritional status. These criteria include a BM < 18.5 kg/mHandgrip strength (HGS) was measured with a hydraulic analogue hand dynamometer (Baseline®) to obtain an indication of peripheral muscle function. Patients were instructed to perform three consecutive contractions with the nondominant hand, and the average power (kg) of the three consecutive measurements was used for analyses. Patients with a HSG below the 10th percentile (P10) of reference values of Bohannon et al. were considered to have a reduced peripheral muscle function .2) and carbon dioxide exhaling (VCO2).Resting energy expenditure was predicted (REE) with the WHO'85 equation based on gender, age, and weight . RestingA ventilated hood system, metabolical monitor , and Weir's equation were used to calculate REE. The measurements were performed in a supine position for 30 minutes in rest, without falling asleep, and patients were instructed to be fasted for at least 12 hours.Patients with a measured REE (mREE) that deviated more than 10% from the predicted REE (pREE) by WHO'85 were considered to have a decreased or increased REE. Predicted total energy expenditure (TEE) was calculated by adding 30% activity factor to REE . To correct the REE for influence of the body composition, REE was divided by REE to calculate the REE in kcal/kg.Patients were instructed to fill out a detailed 4-day food diary, accurately weighing all foods and drinks, using scales. Afterwards, all patients were interviewed by a registered dietitian (JW) to ensure all data was adequately documented. Average intake was calculated using a computerized food calculation program . Total energy intake was determined by adding up the gross energetic value of fat , protein , and carbohydrates  . Total eFasting samples of blood serum were taken at the medical laboratory of the VUmc.μmol/L, vitamin D < 50 nmol/L, and vitamin E < 20 μmol/L. Reference values of international normalized ratio (INR) and activated partial thromboplastin time (APPT) are, respectively, 0.80–1.20 and 25–40 s. Patients with values above these references were suspected for a deficiency of vitamin K.The following vitamin levels were determined: vitamin A (retinol), D (25-OH), E (tocoferol), and the derivatives of vitamin K: international normalized ratio (INR) and activated partial thromboplastin time (APPT). Deficiencies were defined as the following: serum values of vitamin A < 1.2 Assessment of digestion and absorption included analysis of fecal energy and nutrient losses and the fecal elastase-1 (FE1) test.Patients were instructed to collect all 72-hour fecal specimens in a preweighted 5-liter bucket during days 2 to 4 of food intake recording. Patients using pancreatic enzyme replacement therapy (PERT) were instructed to stop using these enzymes at least two days before and during feces collection. All feces collected in 72 hours were weighed, homogenized, and stored at <4°C until analysis. Feces was analyzed for energy, fat, and nitrogen content.A sample of feces was freeze dried until the process of bomb calorimetry, during which the sample was burnt, and the energy produced by the feces was measured as heat of combustion and calculated as fecal energy loss  , 21. TheFecal fat content was determined using the method of van de Kamer et al. . Fecal nThe carbohydrate content was calculated using the following equation /4.10. Finally, the intestinal absorption of energy was calculated as percentage of energy intake by  × 100. Intestinal absorption coefficients of fat, protein, and carbohydrates were calculated similarly , 25. TheExocrine pancreatic function was determined with the fecal elastase-1 (FE1) test.μg/g was defined as abnormal [FE1 was determined from a single feces sample. A FE1 below 200 abnormal .QoL was objectified by the Gastrointestinal Quality of Life Index (GIQLI) questionnaire. This validated questionnaire assesses health-related quality of life of patients with gastrointestinal diseases .Items of the GIQLI questionnaire were scored on a 5-point scale, in which the least desirable and most desirable answer scored, respectively, 0 and 4. The maximum total score was 144. The results were divided into four subscales: physical well-being, mental/physiological well-being, digestion, and defecation .Scores of <55 were considered “unsatisfactory,” ≥55–70 “satisfactory,” ≥70–80 “ample satisfactory,” and ≥80 “very satisfactory.”t-tests and Wilcoxon (Mann–Whitney U) tests were performed to test for statistical differences between variables. A P value < 0.05 was considered to indicate statistical significance. Statistical analyses were performed in Statistical Package for Social Sciences (SPSS) version 22. The results of statistics that capture the performance of a diagnostic test are described as sensitivity (Se), specificity (Sp), and positive predictive value (PPV).Variables were described as mean and standard deviation (SD) if normally distributed and as median and interquartile ranges (IQR) if not normally distributed. Skewness and Kurtosis were used as indicators to test for normality of continuous variables. Independent Information about patient characteristics is shown in The results of the assessment on nutritional status are shown in Based on the new ESPEN criteria for malnutrition, six patients were defined malnourished.Fifty percent (8 patients) had a reduced peripheral muscle function based on a HGS below the 10th percentile.P = 0.002) higher  compared to predicted REE and TEE. Median REE per kg FFM was 34.7 (33.3–49.9) kcal/kg. mTEE was significantly higher  than mean daily energy intake (P = 0.047), but pTEE was not. Mean protein intake was 1.0 g/kg, which is lower than the predefined requirement of ≥1.2 g protein/kg/d, not yet corrected for fecal loss of proteins.Measured REE and TEE were significantly  deficiency in 9/16 patients; none of the patients had used vitamin A, D, or E supplements. None of the patients had an INR or APPT above the reference values.μg/g, whereas thirteen patients showed a FE1 < 200 μg/g.Thirteen out of 15 patients had malabsorption for one or more macronutrients. Out of the five patients that had been prescribed PERT, three had a completely normal fat absorption (>85%) and normal fecal fat excretion (<20 g/d). Besides, 5/15 patients who demonstrated fat malabsorption and increased fecal fat excretion had not been prescribed PERT (data not shown). Median (IQR) FE1 was 61 (21.5–147.5) FE1 had a PPV of 0.54, a Se of 1.0, and a Sp of 0.22 for fecal fat excretion/steatorrhea (>20 g/d). Seven out of 13 patients with a decreased FE1 had no steatorrhea and a normal fat absorption capacity > 85% (6/13).Twelve patients completed the GIQLI . “Very satisfactory” was the most observed score (83%) of QoL. QoL was never assessed as unsatisfactory, once as satisfactory, and once as ample satisfactory. Scores of the total assessment of QoL and scores for each subscale are shown in This study shows that patients with LAPC generally have a severely impaired nutritional status, most likely as a result of an increased REE and malabsorption  due to exocrine pancreatic insufficiency.These results provide an in-depth picture of problems of nutritional status, digestion and absorption capacity, and QoL in patients with LAPC.Although most of the patients had lost a considerable amount of preillness weight in the preceding 6 months, our patients presented mostly with a normal BMI. In contrast, FFMI and peripheral muscle function were low in half (50%) of the patients.In this study, measured REE was significantly higher (33%) than predicted requirements (by WHO'85 equation). This is in line with a previous study which demonstrated an increased REE (33%) in patients with pancreatic cancer compared with healthy controls, suggesting hypermetabolism in patients with advanced pancreatic cancer . HoweverAccording to the estimated energy requirements, energy intake was apparently sufficient. However, according to measured energy requirements, the energy intake was deficient by approximately 423 kcal/d. The median daily protein intake of 1.0 g/kg was generally below the predefined requirements of 1.2 g/kg . HoweverN = 9) of the patients in this small study group suffered from a vitamin D 25-OH deficiency , which is slightly higher than the prevalence of 50% in the “independently living elderly” in The Netherlands [The prevalence of vitamin A and E deficiency was 13% (2/16), most likely due to fat malabsorption in at least half of the patients. However, since the physical storage of fat-soluble vitamins is enough to prevent deficiencies for months to years, sufficient levels of fat-soluble vitamins may be expected . Fifty-sherlands . Since pMalabsorption of energy and macronutrients is an important problem in patients with LAPC. This study showed substantial individual differences in intestinal (energy) absorption capacity, which could not be explained by location of the tumor.Most likely, decreased intestinal absorption capacity was the result of an exocrine pancreatic dysfunction since 13/15 patients had a decreased level of FE1. FE1 is a specific human protease synthesized by the acinar cells of the pancreas. It is stable during transit, and its detection in the stool is uncomplicated . HoweverInterestingly, 5 patients had been prescribed PERT. This proved to be incorrect in 4 (80%), who displayed a normal fat excretion (<20 g/d) and intestinal absorption capacity for fat (>85%). In contrast, 6 patients had not been prescribed PERT, while demonstrating steatorrhea. Furthermore, FE1 as a marker for exocrine pancreatic insufficiency failed to detect steatorrhea as it appeared false positive in 7 out of 15 patients. This suggests that there is room for improvement of prescription of PERT. Based on our study results, PERT should be better prescribed on individual quantitative fecal fat measurements, optionally in combination with FE1, rather than on either clinical presentation, FE1 level, or as a standard procedure.Of the four subscales for QoL, physical and mental well-being scored lower than digestion and absorption. This could be related to a poor nutritional status, low HGS, and low FFMI which may affect functional status and thereby independence and self-reliance of patients. Still, overall quality of life was rated satisfactory or higher in almost all patients. This suggests that patients adapt to their clinical situation and adjust their QoL expectancies. The higher scores for digestion and defecation in this study could be explained by frequently contact with the dietitian with attention for EPI and fecal losses, leading to decrease of clinical symptoms of malabsorption.To our knowledge, this is the first study which extensively assessed nutritional status, digestion, intestinal absorption capacity, and quality of life, combining a broad scala of nutritional assessment techniques in one group of patients with LAPC. A limitation is the relatively small sample size (which was directly linked to the maximum eligible number of patients in the IRE study) and the cross-sectional design. Therefore, the results should be interpreted with caution and no causal links can be drawn.Future research is recommended to assess the nutritional status, digestion and absorption, and QoL prospectively after experimental treatment with IRE.In conclusion, patients with LAPC are at high risk for developing a poor nutritional status and poor physical functioning, most likely due to a combination of increased energy expenditure and malabsorption. Optimizing pancreatic enzyme supplementation in combination with hyperalimentation to correct energy and protein intake for fecal losses may be a strategy to counteract these symptoms as long as patients are actively treated. An individual approach seems necessary given the large differences between patients.


Gynostemma pentaphyllum (GP) in chronic liver disease. One aim of this study was to explore the potential protective effects and mechanisms of action of GP extract on NAFLD. The in vivo results showed that GP extract could alleviate fatty degeneration and haptic fibrosis in NAFLD mice. For exploring the hepatoprotective mechanisms of GP, we used network pharmacology to predict the potential active components of GP and their intracellular targets in NAFLD. Based on the network pharmacology results, we further utilized biomedical assays to validate this in silico prediction. The results showed that Gypenoside XL could upregulate the protein level of PPARα in NAFLD; the transcription level of several PPARα downstream target genes such as acyl-CoA oxidase (ACO) and carnitine palmitoyltransferase-1 (CPT-1) also increased after Gypenoside XL treatment. The overexpression of ACO and CPT-1 may involve the hepatoprotective effects of GP and Gypenoside XL on NAFLD by regulating mitochondrial fatty acid β-oxidation. Nonalcoholic fatty liver disease (NAFLD) is the most common type of liver disease in developed countries. Oxidative stress plays a critical role in the progression of NAFLD. Modern pharmacological study and clinical trials have demonstrated the remarkable antioxidant activity of Oxidd others , 5. Oxidd others –8.Gynostemma pentaphyllum (Thunb.) Makino. (GP), a traditional Chinese medicine, has been reported to have hepatoprotective effect via numerous bioactivities, such as antifibrotic and antioxidative activity [ Jiao Gu Lan in China, the herb mainly contains polysaccharides, flavones, saponins, and trace elements. Recent research has demonstrated that GP extracts and its active components may protect alcoholic fatty liver disease in rats through modulating lipid metabolism and reducing oxidative stress. [activity , 10. GP  stress. , 12. Fuz stress. , 14. One stress. . The mec stress. –17. NAFL stress. –21. Prev stress. , 23. How stress. , 25. In HPLC-MS Analysis of Saponins and Flavonoids Fractions in GP. A total of 10 saponins were separated and identified in GP  and Gypenoside XXIII (339.7 μg/mL) were the most abundant ingredients in total saponins of GP. Likewise, a total of 6 flavonoids were separated and identified  and quercetin-rhamnose (49.3 μg/mL) were the most abundant ingredients in total flavonoids of GP.ed in GP , the totentified ; the totp < 0.01)  . As the μL/kg b.w., twice/week) significantly induced fatty degeneration and hepatic fibrosis in mice liver. The H&E staining results demonstrated that 0.2 and 0.4 g/kg/day GP extracts treatment for 6 weeks significantly attenuated the inflammation and macrovacuolar degeneration in NAFLD  (0.4 in NAFLD . Oil Redin NAFLD . In addiin NAFLD .α) had the largest number (3) of compound connections , followed by BRD4 (Gypenoside XL and quercetin) and SOX9 (Gypenoside XL and Rhamnazin). The other targets (26) showed interactions with just one chemical. Those high-degree nodes with multiple connections in the network, such as Gypenoside XL and PPARα, BRD4, and SOX9, may develop multiple biological effects and play a more pivotal role in treating NAFLD. The data of 29 NAFLD related targets of GP can be obtained from Supplemental The potential active components of GP and their active mechanisms were predicted by in silico method. The drug-target network plotting showed the relationship between GP-derived chemicals and NAFLD related target genes or proteins . The resα protein expression was observed in NAFLD mice (4 exposure can increase their protein expression. Gypenoside XL 10 and 20 mg/kg/day treatment led to a significant increase in PPARα protein level compared with model group (p < 0.05). However, the expression of SOX9 and BRD4 was not affected by Gypenoside XL treatment. CDAA diet and CCl4 exposure can lead to downregulation of ACO and CPT-1 mRNA level as well as protein level in mice liver. Gypenoside XL 10 and 20 mg/kg/day treatment can upregulate mRNA and protein levels of ACO and CPT-1 , a key node in our network pharmacology prediction, can modulate lipid peroxidation in fatty liver disease. PPARα is a kind of nuclear receptor proteins; it is an important transcription factor for regulating the key enzymes in beta-oxidation pathway. Previous studies showed that PPARα expression was downregulated in ALD and NAFLD in human liver [α can inhibit fatty liver disease by activating the periostin-dependent JNK signaling pathway and modulating fatty acid oxidation [α in liver can also prevent acetaminophen-induced liver damage by upregulating mitochondrial glutathione and downregulating fatty acyl-carnitines concentration in blood [α also protect liver tissue by activating fatty acid beta-oxidation related pathway [α might be an important target for preventing NAFLD. Previous studies showed that several ingredients in GP such as Gypenosides may target PPARα and regulate the expression of downstream genes in human endothelial cells and monocytic cells [α in NAFLD liver tissue. Although PPARα has no direct effects to haptic lipid metabolism, it can up-regulate the expression of some key genes in mitochondrial and fatty acid β-oxidation [α expression can induce the transcription of β-oxidation related enzymes, the transcription level of several PPARα downstream target genes such as acyl-CoA oxidase (ACO) and carnitine palmitoyltransferase-1 (CPT-1) was analyzed by RT-PCR and Western blot. Previous studies have shown that the expressions of ACO and CPT-1 were suppressed in NAFLD liver tissue, which is consistent with our results [α expression may relate to the hepatoprotective effects of GP and Gypenoside XL on NAFLD.Peroxisome proliferator-activated receptor alpha , and has been authenticated by Dr. Zhe Cai . A voucher sample of the whole herb (SCM 127) was deposited at the School of Chinese Medicine, Faculty of Medicine, HKU, and the whole herb of Gynostemma pentaphyllum was cleaned and sliced into small pieces, after soaking for 3 hour and boiling for 1.5 hours under a high-heat fire. The filtered herbal residue was boiled again by the same method in above step. Then, all filtrate was collected, combined, and then concentrated at 60°C double distilled water to a final concentration of 0.5 g/mL. Gypenoside XL (PubChem CID: 94705-70-1) powder was purchased from Sigma-Aldrich Company in USA. Gypenoside XL was dissolved in Distilled Deionized Water and diluted in different concentration.Choline-deficiency amino acid-defined diet (CDAA) and control chow diet were obtained from Research Diets ; carbon tetrachloride (CCl4) was purchased from Sigma-Aldrich ; detection kits for alanine transaminase (ALT), aspartate transaminase (AST), malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GPx) are purchased from Jianchen Company ; direct red 80 and picric acid were purchased from Sigma-Aldrich .A method as described by Kao et al. was modified to prepare saponins and flavonoids from GP by open-column chromatography . The varμL/kg b.w., twice/week) was used as promoter of hepatic fibrosis [For the establishment of NAFLD model, mice were fed with control chow or CDAA chow for 6 weeks. A low dose of carbon tetrachloride (CCl4) . Fresh liver tissue was sliced into small piece and homogenized in lysis buffer, centrifuged at 12,000 ×g for 5 min at 4°C, and the supernatants were collected and stored at −80°C until biochemical analysis. The activity of GSH-Px and SOD and the production of MDA were measured according to the manufacturers' instructions .For hematoxylin and eosin staining, fresh mice liver tissue was sliced into 1.5 × 1.5 cm; then the tissues were fixed by formalin. The slides containing paraffin coated tissues were placed in a glass holder; then the slides were deparaffinized and rehydrated by different concentration of xylene and ethanol; then the slides were stained by hematoxylin for 5 minutes; after tab water washing and eosin staining for 30 seconds, the slides were dehydrated by different concentration of xylene and ethanol. All slides were covered by xylene based permount and stored in cool and dry environment. The histological results were analyzed by microscopy. The liver damage score was analyzed by two individual examiners with the following criteria: 0, no visible damage; 1–3, mild damage; 4–6, intermediate damage; 7–9, severe damage; and 10, destruction of hepatic structure. For assessing liver fibrosis, we have conducted Picrosirius red staining in hepatic tissue biopsy. The fresh liver tissue was fixed in formaldehyde solution for 48 hours. After paraffin embedding, the paraffin sections were dewaxed and hydrated by ethanol. Then, nucleus was stained with hematoxylin for 5 minutes, and then we used running tap water to wash the slides for 5 minutes. We stained the slides in Picrosirius red for 60 minutes and washed the slides in acidified water. Then, we shook the slides and dehydrated them in ethanol. Finally, the biopsy was mounted in a resinous medium and analyzed by microscopy. Hepatic fibrosis scoring on the stained sections was made by two individual examiners with the following criteria: 0, no signs of observed fibrosis; 1–3, no extension of portal area fibrosis; 4–6, fibrosis occurring in the portal area with an intact lobule structure; 7–9, fibrosis associated with a broken lobule structure, but no signs of cirrhosis; and 10, fibrosis and the formation of cirrhosis. The liver steatosis scores were evaluated by Oil Red O staining. Briefly, cryosectioned livers were washed with running tap water for 10 minutes, rinsed with 60% isopropanol, and stained with Oil Red O  mixed with 60% isopropanol for 15 minutes. The sections were then rinsed with 60% isopropanol, and nuclei were stained with Mayer's Hematoxylin, rinsed in tap water, and coverslipped with aqueous mounting medium. The liver steatosis score was analyzed by two individual examiners with the following criteria: 0, no visible steatosis; 1–3, mild steatosis; 4–6, intermediate steatosis; and 7–10, severe steatosis.http://tcm.cmu.edu.tw/) and Traditional Chinese Medicine Systems Pharmacology Database (TCMSP) (website: http://lsp.nwu.edu.cn/tcmsp.php). The inconsistent data in these two databases were further verified and confirmed by related literatures review.The chemical ingredients of GP were collected from following phytochemical databases: TCM Database@Taiwan (website: http://lsp.nwu.edu.cn/tcmsp.php). At last, ingredients meet the case that Caco2 ≥ 0.4 cm/s and OB ≥ 33% were chosen as candidate compounds for function analysis in the next step.A computer-based integrative model, ADME , was applied in this step to screen the chemicals with eligible pharmacokinetics properties in GP. PreOB  and PreCaco-2 (prediction of Caco-2 permeability) were major pharmacokinetics properties indicator in our ADME screening model. Oral bioavailability (OB) refers to the efficiency of the drug delivery to the blood circulation, which is an important property for oral medications. For calculating the OB value of GP-derived single compounds, OBioavail 1.1, a robust in silico model designed by Northwest A&F University, was applied in this step. This model was designed based on more than 800 structurally diverse drugs and drug-like agents. Support vector machine, multiple linear regression, and partial least square methods were used for building this model; the detailed algorithms of this model can be found in previous studies by Wang et al. . Beside http://crdd.osdd.net/raghava/npact/) and Search Tool for Interactions of Chemicals and Proteins (STITCH) database (website: http://stitch.embl.de/); the information of ingredients and intracellular targets interactions retrieved from these databases was integrated and saved for further study. Then, for providing a more focused view on compound-NAFLD target interaction, we eliminated the unrelated target protein associations by mapping these targets to the Therapeutic Target Database (TTD) (website: http://bidd.nus.edu.sg/group/cjttd/). The above databases provide detailed information of the known or explored chemical agents and the corresponding intracellular targets and related human disease and pathologic changes. The protein targets related to NAFLD were further analyzed and confirmed by PharmGKB (website: http://www.pharmgkb.org) through using the following search terms: oxidative stress, fatty liver, hepatic steatosis, nonalcoholic liver disease, liver fibrosis, oxidative damage, and lipid metabolism.The biological activity of each single compound in GP was searched and analyzed using the software of Herbal Ingredients' Targets (HIT) Database (website: http://www.cytoscape.org/) was used to construct the compound-target network plotting. In this visible network plot, a node stands for a chemical compound, gene or protein; the edge represents the compound-target interaction. The information of different nodes and their connections in the network were further analyzed by Network Analysis plugin to determine the key nodes for further experimental validate. The key nodes present the potential pivotal compounds or target proteins which may play important role in preventing NAFLD. To provide a visible network between the GP-derived ingredients and NAFLD related protein targets, the most update Cytoscape 3.4.0  at 4°C overnight, then, after cleaning the membrane with TBST for 15 minutes for 3 times, the membrane was further incubated with corresponding secondary antibodies at room temperature for 3 hours. The immunoreactivities were tested by using Electrochemiluminescence Advanced kit and the results were analyzed with Bio-Rad chemi-luminescence imaging system. The protein expression levels of PPARμM primers  in LightCycler 480 real-time PCR machine. The mRNA expression of β-actin  was used as endogenous control. The relative expression of CPT-1, ACO mRNA for each sample was normalized with β-actin and the relative value was compared with normal control mice.Total RNA from liver tissue was extracted and purified using RNeasy Mini Kit following the manufacturer's instruction. QuantiTect Reverse Transcription Kit was used for reverse transcription of total RNA. The quantitative real-time PCR was conducted by using LightCycler 480 SYBR Green I master with 1 t-test. Value of p < 0.05 was considered to be statistically significant.All the data obtained are presented as the mean ± SD and analyzed by one-way analysis of variance (ANOVA) with Statistical Product and Service Solutions (SPSS) version 19 . The significance of two groups was examined using a Student's


Hypertension and the triglyceride and glucose index both have been associated with insulin resistance; however, the longitudinal association remains unclear. This study was designed to investigate the longitudinal association between the triglyceride and glucose index and incident hypertension among the Chinese population.We studied 4686 subjects  and followed up for 9 years. The subjects were divided into four groups based on the triglyceride and glucose index. Univariate and multivariate Cox regression models were used to analyse the risk factors of hypertension.p for trend < 0.001). Cox regression analyses indicated that higher triglyceride and glucose index was associated with an increased risk of subsequent incident hypertension.After 9 years of follow-up, 2047 subjects developed hypertension. The overall 9-year cumulative incidence of hypertension was 43.7%, ranging from 28.5% in quartile 1 to 36.9% in quartile 2, 49.2% in quartile 3 and 59.8% in quartile 4 (The triglyceride and glucose index can predict the incident hypertension among the Chinese population. With the population aging, hypertension (HTN) has become the most important risk factor for cardiovascular disease (CVD) morbidity and mortality –4. AccorRecently, the triglyceride and glucose (TyG) index has been proposed as a simple surrogate of insulin resistance (IR) , 9, and The study participants, aged 20–80 years, were enrolled in the annual physical health examinations at Zhenhai Lianhua Hospital in Ningbo, China. This longitudinal population-based cohort study was conducted to investigate the longitudinal association between the TyG index and the incident HTN beginning from 2006 to 2015. The subjects were excluded according to the following criteria: (1). Subjects who had a history of HTN or overt cardiovascular diseases. (2). Subjects who were taking medicines that may affect blood pressure or lipids. (3). Subjects who were drinking alcohol and smoking at study entry. In total, 4686 subjects including 3177 males and 1509 females were evaluated for the study. A questionnaire was completed for the initial health examinations in 2006 at the health check-up center of Zhenhai Lianhua Hospital through face-to-face interviews by a well-trained senior physician. The questionnaire included the subjects’ demographic characteristics, smoking and drinking status, medical history, and family background. Anthropometric test: Standing height and body weight was measured while he/she was in light indoor clothing without shoes. Body mass index (BMI) was calculated as weight in kilograms divided by height in meters squared. Waist circumference (WC) was measured at the level midway between the lower rib margin and the iliac crest . Sitting Laboratory examinations: Venous blood samples were obtained from the subjects in the morning after at least 12 h prior to the examination. Routine biochemical data including blood urea nitrogen (BUN), creatinine (Cr), triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein (LDL-C), serum uric acid (SUA), fasting plasma glucose (FPG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyltransferase (γ-GGT), Apo-A1and Apo-B were estimated using an Olympus AU640 automated analyser . All of the laboratories involved resoundingly completed the standardization. HTN was defined as systolic blood pressure (SBP) ≥ 140 mmHg, diastolic blood pressure (DBP) ≥ 90 mmHg or current drug use for HTN in accordance with the criteria of the WHO [ The TyG index was calculated with established formulas according to the previous studies , 16: TyG The estimated glomerular filtration rate (eGFR) was calculated using the improved Chinese population Modification of Diet in Renal Disease (MDRD) study formula .U test, Kruskal-Wallis H test or one-way ANOVA depending on the normality of the data. Categorical variables between groups were compared using Chi-square text. For a statistical inference, all p values are bilateral, and a p value of less than 0.05 was considered statistically significant.The fundamental characteristics of the samples were summarized by descriptive statistics. All statistical analyses were performed using SPSS software . Continuous variables were expressed as median (IQR) and categorical variables were presented as percentages (%). Continuous variables were compared using the student’s t text, Mann-Whitney The subjects were classified into four groups (quartile 1 to quartile 4) based on the TyG index at baseline. The classifications of the TyG index in these groups were as follows: quartile 1(Q1) (≤ 8.12), quartile 2(Q2) (8.13–8.46), quartile 3(Q3) (8.47–8.83), and quartile 4(Q4) (≥ 8.84) for male; and quartile 1(Q1) (≤ 7.85), quartile 2(Q2) (7.86–8.16), quartile 3(Q3) (8.17–8.52), and quartile 4(Q4) (≥ 8.53) for female. The basic characteristics of the subjects in each group were compared. The cumulative incidence of HTN was calculated by dividing the number of cases by the numbers of subjects followed up for each TyG group. Cox proportional hazards regression models were used to analyze the risk of incident HTN for each baseline TyG group.p < 0.001). In addition, age, BMI, SBP, DBP, WC, FPG, UA, AST, ALT, γ-GGT, TC, TG, LDL-C and Apo-B all tended to increase in subjects with a higher TyG index, whereas HDL-C and eGFR were significantly lower in subjects with a higher TyG index (p < 0.001).The basic demographic and clinical characteristics of the subjects are presented in Table p for trend < 0.001; Fig. To investigate the predictive value of the TyG index for incident HTN, our longitudinal study was conducted for 9 years. After a 9-year follow-up, 2047 subjects  developed HTN, corresponding to 48.5 and 33.5% cumulative incidence of HTN in males and females, respectively. Figure p for trend < 0.001). The same relationship between the TyG index and incident HTN was also revealed even after adjusting for age and gender (Mode 1); or age, gender, and BMI (Mode 2); or age, gender, and all clinical variables (Mode3) in Table We used univariate and multivariate Cox proportional hazard models to analyse the hazard ratio for incident HTN in each TyG group in our study Tables  and 4. CThis population-based study demonstrates that there is a longitudinal relationship between the TyG index and the risk of incident HTN during a 9-year period among the Chinese population. We also found that the TyG index is an independent predictor for incident HTN. Cox regression analysis suggests that subjects with a higher baseline TyG index are significantly associated with a higher risk of incident HTN after adjusting for the confounders. This study also confirms that the findings of relevant cross-sectional studies, which observe an independent positive relationship between the TyG index and incident HTN .2O and Na+ indirectly, cause water-sodium retention and increase the vascular activity with noradrenaline and AT-II, leading to the HTN eventually [IR has been the significant risk factor for the development of HTN , 18 and entually , 22. Slaentually  also fouentually , 25, andPrevious epidemiological studies have shown that the TyG index was a well-known predictor for the development of diabetes mellitus . Our resThe major strengths of this study were the 9-year longitudinal population-based study and the large number of the subjects. This longitudinal study also expanded the observation to establish the temporal sequence between the TyG index and the later risk of HTN in China. Moreover, the selection bias was less likely to appear in the present study as annual health check-ups in state-owned companies are mandatory in China. However, certain limitations also exist in this study. First, fasting insulin was not obtained due to the lack of relevant devices. Second, we did not record the information on nutritional habits or energy intake in this study. Although we did not adjust for this possible confounding factor, we used other additional covariates indirectly related to nutritional habits, such as BMI or cholesterol. Third, taking lipid-lowering medications or nutraceuticals might have influenced the lipids or the TyG index values. However, the percentage of subjects on these agents was towards the null, we did not think that it would influence the results. Fourth, physical activity may affect the blood pressure or lipids. However, the duration and intensity of our subjects’ physical activity in this study was almost similar, we did not think it would affect the results either. Finally, all subjects in the present study were enrolled from one hospital, so the conclusions may differ from the general population. Given these limitations, further studies should be carried out to clarify the above factors.In conclusion, our study showed that the TyG index independently predicted the incident HTN. These findings suggested that the TyG index should be introduced to the routine check-ups, which could benefit for the prevention of HTN.


B. longum 1714 strain as a putative psychobiotic with an impact on stress-related behaviors, physiology and cognitive performance. Whether such preclinical effects could be translated to healthy human volunteers remains unknown. We tested whether psychobiotic consumption could affect the stress response, cognition and brain activity patterns. In a within-participants design, healthy volunteers (N=22) completed cognitive assessments, resting electroencephalography and were exposed to a socially evaluated cold pressor test at baseline, post-placebo and post-psychobiotic. Increases in cortisol output and subjective anxiety in response to the socially evaluated cold pressor test were attenuated. Furthermore, daily reported stress was reduced by psychobiotic consumption. We also observed subtle improvements in hippocampus-dependent visuospatial memory performance, as well as enhanced frontal midline electroencephalographic mobility following psychobiotic consumption. These subtle but clear benefits are in line with the predicted impact from preclinical screening platforms. Our results indicate that consumption of B. longum 1714 is associated with reduced stress and improved memory. Further studies are warranted to evaluate the benefits of this putative psychobiotic in relevant stress-related conditions and to unravel the mechanisms underlying such effects.The emerging concept of psychobiotics—live microorganisms with a potential mental health benefit—represents a novel approach for the management of stress-related conditions. The majority of studies have focused on animal models. Recent preclinical studies have identified the  A promising strategy in this field is that of psychobiotics—live microorganisms that convey a benefit upon the host's mental health when consumed in adequate quantities.9 a stress-related brain–gut axis disorder associated with high rates of psychopathology10 as well as altered hypothalamic–pituitary–adrenal axis activity11 and cognition.13 A number of encouraging proof-of-principle studies in healthy human volunteers have now demonstrated that multistrain probiotics, fermented drinks containing probiotics or prebiotics can alter resting brain activity, cognitive performance, baseline physiological stress outputs and self-reported psychological variables.18 It is, however, unclear whether the use of single putative psychobiotic strains is also a viable approach in humans and whether a discovery pipeline can be developed to inform superior candidate strain selection as well as evaluation across a broader range of relevant parameters.Although most of the evidence in this area comes from animal studies, specific probiotic strains have shown potential for symptom alleviation in irritable bowel syndrome (IBS),1 The rational selection of candidate strains identified in well-validated preclinical screening platforms is a more logical approach to guide the challenging move toward clinically useful psychobiotics. Accordingly, in preclinical studies, we have identified the Bifidobacterium longum 1714 strain, which selectively improves stress-related behaviours, physiology and cognitive performance.20 The current study investigated whether these preclinical effects could be translated to healthy human volunteers.We have recently proposed a precision strategy in the use of psychobiotics for targeting stress-related central nervous system disorders including anxiety and depression.22 We also assessed cognitive performance; in addition to our previous preclinical findings indicating improved memory, there is existing evidence that consumption of probiotics can affect performance on sustained attention performance,18 as well as in brain activity during social cognition.15 We thus assessed cognitive performance on tests assessing memory, sustained attention, social cognition and emotional processing. We also assessed brain activity in frontal, parietal and central regions using electroencephalography (EEG) following 4-week supplementation with the 1714 strain in comparison with placebo, as these regions have been associated with memory and sustained attention24 and are sensitive to anxiolytics.25We examined the effects of the 1714 strain compared with placebo on daily reported stress and the psychobiological response to an acute, controlled stressor.The research described received approval from the Clinical Research Ethics Committee of the Cork Teaching Hospitals (Protocol Number: APC044). Informed consent was obtained from all participants, who were free to withdraw from the study at any time.26), and demographic and baseline psychological information was collected. Following screening, participants completed neurocognitive visits and acute stress visits. Participants were then administered placebo for four weeks followed by the 1714 strain for four weeks, with the second and third set of visits following the placebo and probiotic phases. Participants also completed a 2-week post-probiotic follow-up. Participants filled in daily online questionnaires from the end of the first set of visits to the end of follow-up.A repeated measures, placebo-controlled design was employed. A repeated measures design was employed in order to exclude the effects of individual differences across variables. Participants were screened at an initial visit for psychiatric disorder using the MINI International Neuropsychiatric Interview  were presented on a touch-screen monitor, Sahara i440D Slate Tablet PC  running CANTABeclipse software . The researcher provided verbal instructions to participants from a standardised script. Tests were presented in different orders for different participants, using a Latin square design, to avoid effects of fatigue for tests completed later in the session. The test battery lasted ~45 min in total. The Paired Associate Learning (PAL) test was used to assess conditional learning of pattern–location associations. Paired associate learning performance has shown sensitivity to functional changes in the hippocampus31 and frontal lobes.23 The parallel mode (which presents different shapes at each visit) was used in order to avoid practice effects.Tests from the Cambridge Neuropsychological Test Automated Battery  and frequency of use.36In addition to tests from the CANTAB battery, we assessed emotional processing using an Emotional Stroop.25 See We used an auditory oddball task to study the P300 event-related potential, which has been shown to be sensitive to anxiolytic effects at the cerebral electrode locations assessed in the current study.Prior to EEG testing participants were asked to refrain from caffeine on the morning of their experimental session, as well as ensuring they got a good night's sleep, to remove any piercings and avoid wearing hair gel. All EEG measurements were made using a Neuroscan, SynAmps 2 Amplifier and Neuroscan 4.3.1 acquisition software . EEG data were recorded at a sampling rate of 1000 Hz. Scalp electrodes were attached at midline positions Fz, Pz and Cz, according to the international 10/20 system, as well as mastoid electrodes and a reference electrode on the nose. Vertical eye movements were detected using electrodes attached above and below the orbit of the left eye and horizontal eye movements were monitored by electrodes at the right and left outer canthi. Electrodes were applied to the scalp using soft paste and secured using tape and an elasticated hat.Following a resting EEG recording, the cognitive tasks were completed (see Cognitive tasks). EEG was recorded continuously during performance of the auditory oddball task, but was not measured during the other cognitive tasks to avoid artefacts in the EEG readings.37). Participants were requested to relax and sit still with their eyes closed while resting EEG was recorded. Resting EEG has been shown to be consistent across repeated visits,38 and so should not be vulnerable to carryover effects.EEG measures of absolute power in the delta (1.5–3.5 Hz), theta (4–7.5 Hz), alpha1 (8–9.5 Hz), alpha2 (10–12.5 Hz), beta1 (13–17.5 Hz) and beta2 (18–25.5 Hz) frequency bands were taken for an initial five minute period with eyes closed Resting EEG, frequency domain Data were examined for eyeblink artefacts. After correction for eyeblinks, data were epoched, with an epoch duration of 2 s. A linear detrend based on the entire sweep was applied to the epoched file, which was then baseline corrected for the pre-stimulus interval. A band pass filter  was applied. Following a further visual inspection for artefacts and baseline correction the epochs were averaged. Multiplication of data by a 25% cosine window was applied, as this has been used previously in examining anxiolytic effects.41 EEG measures of absolute power were extracted in the delta (1.5–3.5 Hz), theta (4–7.5 Hz), alpha1 (8–9.5 Hz), alpha2 (10–12.5 Hz), beta1 (13–17.5 Hz) and beta2 (18–25.5 Hz) frequency bands.Resting EEG, time domain The EEG signal was downsampled from 1000 Hz to 256 Hz with an antialiasing filter set at 128 Hz. The filtered EEG signal was segmented into 1 s window without overlap. Curve length, number of maxima and minima, root mean squared amplitude, Hjorth parameters 42 , zero crossings , autoregressive modelling error (model order 1–9), nonlinear energy, variance  were calculated using MATLAB. Mobility is an estimate of root-mean-square frequency. Mobility=std(y(t)dy/dt)/(y(t)), where the y(t) is the EEG amplitude at time, t.Event-related potential Data were divided into epochs of 300 ms duration . Following a linear detrend and a baseline correction, data were visually examined for any artefacts and these were removed. Oddball epochs were averaged, and a band pass filter and further baseline correction were applied. The peak frequency was identified between 200 ms and 500 ms post-oddball for the P300.−1. Inter and intra-assay coefficients of variability were 11.24% and 8.2%, respectively.Salivary cortisol was analysed using Enzo Life Sciences  enzyme-linked immunosorbent assay (ELISA) kits  according to manufacturer's instructions. Lower limit of detection=0.16 nmol lt-tests (two-sided) were used to examine differences between conditions, and non-parametric equivalents  were used where parametric assumptions were violated. Areas under the curve with respect to ground (AUCg) were also calculated,43 and analysed in the same manner. EEG data for three participants were removed (two had excessive levels of artefacts and one participant suffered a tension headache when the EEG equipment was attached). Daily questionnaire data for three participants were removed due to lack of response to online questionnaires.Data were analysed using SPSS 21 . Repeated measures analysis of variance and T=5, P<0.001, r=0.8, post-placebo: T=4.67, P<0.001, r=0.78, post-1714: T=2, P<0.001, r=0.94), but there was not an interaction between probiotic condition and stressor time point, F=1.81, P=0.18 =8.67, P<0.05, although area under the curve with respect to increase (AUCi) was not affected, χ2(2)=3.71, P>0.05  compared with immediately pre-stress (time 0) for all three conditions =6.39, , r=0.12 . The meat(18)=1.95, P=0.07, Cohen's d=0.44. Further, stress levels returned to a higher level during the 2-week follow-up period (t(18)=2.32, P=0.03, Cohen's d=0.53, and AUCi was lower in the 1714 condition, although this was not statistically significant, t(18)=1.44, P>0.05, Cohen's d=0.33 =10.46, P<0.01. Compared with the first visit, participants made fewer errors on the paired associate learning test both at post-placebo,  and post-1714, , a subtle effect at post-1714 that was greater than the placebo response =13.37, P=0.01 =10.31, P<0.01. Cz theta power was significantly lower post-1714 compared with post-placebo, T=8, P<0.05, r=0.57, although theta power at post-1714 did not differ from visit one  mobility using EEG. Activity at Fz is representative of prefrontal cortex activity,44 and the prefrontal cortex is associated with paired associate learning.23 As prefrontal activity appears to be specifically associated with top–down processing during associative memory,23 the current EEG findings thus offer a potential insight into the timing of cognitive processes underlying the observed changes in paired associate performance, by suggesting that the impact of the psychobiotic on cognitive performance may be due to an effect on top–down processing, rather than simply affecting retention. Mobility has previously been shown to be higher in healthy controls compared with patients with Alzheimer's disease,45 a neurodegenerative disorder with pronounced and progressive memory deficits.46 Although Alzheimer's disease is a severe clinical condition, it is worth noting that the observed effects in Alzheimer's patients were stronger frontocentrally, which is where the change in mobility was observed post-psychobiotic. The reduction in theta power post-psychobiotic compared with post-placebo is also consistent with improved memory performance.47The subtle improvement in visuospatial memory following probiotic administration is consistent with preclinical findings of enhanced learning and memory in response to this strain.48 which has a high proportion of glucocorticoid receptors,49 and PAL test performance is poorer in subjects with stress-related disorders with associated hypothalamic–pituitary–adrenal axis dysfunction, such as IBS.13B. longum NCC3001 enhanced hippocampal brain-derived neurotrophic factor messenger RNA and reduced anxiety-like behavior in mice infected with the non-invasive parasite T Muris50 and B. breve 6330 increased brain-derived neurotrophic factor total variants in healthy rats.51 It is thus of interest whether brain-derived neurotrophic factor levels and neurogenesis are heightened in humans following psychobiotic ingestion, thus having a key role in the impact of psychobiotic consumption upon the central nervous system. Furthermore, given previous evidence that psychobiotic effects in rodents were dependent upon the vagus nerve,7 it is possible that vagal activity may mediate such psychobiotic effects. However, strain-specific mechanisms are likely and need to be investigated on a case by case basis.Further research is warranted to further elucidate mechanisms by which the 1714 strain could impact upon stress and neurocognition. It is possible that the reduction in daily stress and acute stress response may account for the observed effect on visuospatial memory. PAL test performance is dependent upon the hippocampus,13 and we have shown PAL performance to be subtly improved by the 1714 strain. Furthermore, the observed psychological effects of psychobiotics warrant research in other stress-related disorders including depression and anxiety.These results provide the impetus for further evaluation of this psychobiotic in pathological conditions. This could initially target stress-related brain–gut axis disorders with a cognitive component such as IBS, which has been found to be associated with a deficit in visuospatial memory performance, evident in PAL test performance,In line with the predicted impact from preclinical research in this specific strain, the 1714 strain is a promising candidate psychobiotic strain associated with attenuated responses to psychological and physiological stress and a modest improvement over placebo in cognitive performance, as well as with altered resting EEG output in healthy volunteers. Further studies are warranted to evaluate the benefits of this putative psychobiotic in relevant stress-related conditions and to unravel the mechanisms underlying such effects.