Clinical efficacy of the co-administration of Turmeric and Black seeds (Kalongi) in metabolic syndrome - a double blind randomized controlled trial - TAK-MetS trial.

OBJECTIVE: To compare the clinical efficacy of Black seeds and Turmeric alone and its co-administration in lower doses among patients with metabolic syndrome (MetS). DESIGN: Double-blind-randomized-controlled trial. SETTING: Hijrat colony, Karachi, Pakistan. INTERVENTION: Apparently healthy males (n=250), who screened positive for MetS, were randomized to either Black seeds (1.5g/day), Turmeric (2.4g/day), its combination (900mg Black seeds and 1.5g Turmeric/day) or placebo for 8 weeks. MAIN OUTCOME MEASURES: body-mass-index (BMI), body-fat-percent (BF%), waist-circumference (WC), hip-circumference (HC), blood pressure (BP), lipid-profile (cholesterol, HDL-cholesterol, LDL-cholesterol and TG), fasting blood glucose (FBG) and c-reactive protein (CRP). RESULTS: At 4 weeks, compared to baseline, Black seed and Turmeric alone showed improvement in BMI, WC and BF%. Combination improved all parameters except HDL-cholesterol with lower FBG and LDL-cholesterol as compared to placebo. At 8 weeks, compared to placebo, Black seeds reduced lipids and FBG, while Turmeric reduced LDL-cholesterol and CRP. Interestingly, combination group with 60% dose of the individual herbs showed an improvement in all parameters from baseline. When compared to placebo, it reduced BF%, FBG, cholesterol, TG, LDL-cholesterol, CRP and raised HDL-cholesterol. CONCLUSION: Turmeric and Black seeds showed improvement in all parameters of metabolic syndrome, when co-administered at 60% of doses of individual herbs with enhanced efficacy and negligible adverse-effects. The combination of Black seeds and Turmeric can therefore, be recommended with lifestyle modification as a starting point for patients with MetS to halt its future complications and progression.

Gut modulator effects of Conyza bonariensis explain its traditional use in constipation and diarrhea.

BACKGROUND AND OBJECTIVES: The current investigation was carried out to explore the pharmacological basis of the crude extract of Conyza bonariensis (Cb.Cr) for its use in constipation and diarrhea. MATERIALS AND METHODS: The plant extract of Conyza bonariensis (C. bonariensis) was prepared, isolated guinea-pig ileum and rabbit jejunum preparations were used to evaluate its gut modulator effects. RESULTS: The Cb.Cr (0.3-10 mg/mL) exhibited spasmogenic effect in isolated guinea-pig ileum preparation, which was about 19-84% of the acetylcholine maximum. Pretreatment of the tissues with atropine (0.1 µM) abolished the contractile effect, similar to acetylcholine. Among the fractions, only the butanol fraction exhibited atropine sensitive contractile effect. In isolated rabbit jejunum preparations, Cb.Cr produced appreciable atropine-sensitive spasmogenic effect at lower concentrations (0.03-0.3 mg/mL) followed by spasmolytic effect at next higher concentration (1.0 and 3.0 mg/mL). Cb.Cr caused an inhibition of the high K+ induced contraction in isolated rabbit jejunum preparation with EC50 value of 0.62 mg/mL. Similarly, verapamil, a standard calcium blocker, inhibited high K+ induced contraction in isolated rabbit jejunum preparations. Cb.Cr caused a right ward shift in the Ca++ concentration response curve, similar to verapamil. Among various fractions of C. bonariensis, only hexane and ethylacetate fractions showed spasmolytic effects. CONCLUSIONS: The crude extract of C. bonariensis contains spasmogenic and spasmolytic constituents, which explains its medicinal use in constipation and diarrhea.

The analgesic and anticonvulsant effects of piperine in mice.

Piperine, is the major active principal of black pepper. In traditional medicine, black pepper has been used as an analgesic, anti-inflammatory agent and in the treatment of epilepsy. This study was conducted to evaluate the in vivo analgesic and anticonvulsant effects of piperine in mice. The analgesic and anticonvulsant effects of piperine were studied in mice using acetic acid-induced writhing, tail flick assay, pentylenetetrazole (PTZ)- and picrotoxin (PIC)-induced seizures models. The intraperitoneal (i.p.) administration of piperine (30, 50 and 70 mg/kg) significantly inhibited (P<0.01) the acetic acid-induced writhing in mice, similar to the effect of indomethacin (20 mg/kg i.p.). In the tail flick assay, piperine (30 and 50 mg/kg, i.p.) and morphine (5 mg/kg, i.p.) caused a significant increase (P<0.01) in the reaction time of mice. Pre-treatment of animals with naloxone (5 mg/kg i.p.), reversed the analgesic effects of both piperine and morphine in the tail flick assay. Piperine (30, 50 and 70 mg/kg, i.p.) and standard drugs, valproic acid (200 mg/kg, i.p.), carbamazepine (30 mg/kg, i.p.) and diazepam (1 mg/kg, i.p.) significantly (P<0.01) delayed the onset of PTZ-and PIC-induced seizures in mice. These findings indicate that piperine exhibits analgesic and anticonvulsant effects possibly mediated via opioid and GABA-ergic pathways respectively. Moreover, piperine being the main constituent of black pepper, may be contributing factor in the medicinal uses of black pepper in pain and epilepsy.

Curcuminoids enhance memory in an amyloid-infused rat model of Alzheimer's disease.

Alzheimer's disease (AD) is a neurodegenerative disease. There are a limited number of therapeutic options available for the treatment of AD. Curcuminoids (a mixture of bisdemethoxycurcumin, demethoxycurcumin and curcumin) is the main chemical constituent found in turmeric, a well known curry spice, having potential in the treatment of AD. The objective of this study was to investigate the effects of curcuminoid mixture and individual constituents on spatial learning and memory in an amyloid-beta (Abeta) peptide-infused rat model of AD and on the expression of PSD-95, synaptophysin and camkIV. Curcuminoid mixture showed a memory-enhancing effect in rats displaying AD-like neuronal loss only at 30 mg/kg, whereas individual components were effective at 3-30 mg/kg. A shorter duration treatment with test compounds showed that the curcuminoid mixture and bisdemethoxycurcumin increased PSD-95 expression in the hippocampus at 3-30 mg/kg, with maximum effect at a lower dose (3 mg/kg) with respective values of 470.5 and 587.9%. However, after a longer duration treatment, two other compounds (demethoxycurcumin and curcumin) also increased PSD-95 to 331.7 and 226.2% respectively at 30 mg/kg. When studied for their effect on synaptophysin in the hippocampus after the longer duration treatment, the curcuminoid mixture and all three individual constituents increased synaptophysin expression. Of these, demethoxycurcumin was the most effective showing a 350.1% increase (P<0.01) at 30 mg/kg compared to the neurotoxin group. When studied for their effect on camkIV expression after longer treatment in the hippocampus, only demethoxycurcumin at 30 mg/kg increased levels to 421.2%. These compounds salvaged PSD-95, synaptophysin and camkIV expression levels in the hippocampus in the rat AD model, which suggests multiple target sites with the potential of curcuminoids in spatial memory enhancing and disease modifying in AD.

In vivo anthelmintic activity of Azadirachta indica A. Juss seeds against gastrointestinal nematodes of sheep.

This paper describes the in vivo anthelmintic activity of Azadirachta indica seeds to justify their use in South-Asia by traditional animal healers. Seeds of A. indica were administered as crude powder (CP), crude aqueous (CAE) and crude methanolic extracts (CME) at the doses of 1 and 3g/kg of body weight to sheep naturally infected with mixed species of gastrointestinal nematodes. The study design also included untreated as well as treated controls. Faecal egg count reduction and larval counts from coprocultures were performed pre- and post-treatments to assess the anthelmintic activity. Crude powder and CME did not show significant activity (P>0.05) at the lower dose used but were found effective at 3g/kg and the maximum anthelmintic effect was observed at the 15 days post-treatment with both crude powder and CME (P<0.01) with a maximum reduction of 29.3% and 40.2%, respectively in eggs per gram of faeces. Haemonchus contortus and Trichostrongylus species were found susceptible (P<0.01) to higher doses of CP and CME of A. indica. However, CAE did not exhibit any considerable reduction in EPG as well as larval counts. Levamisole (7.5mg/kg), a standard anthelmintic agent, exhibited 99.2% reduction in EPG (P<0.001). Though of low efficacy compared with levamisole, the use of A. indica seeds against gastrointestinal nematodes may be justified in some situations, depending on the nature and intensity of the helminth infections.

Cardiovascular inhibitory effects of Hyoscyamus niger.

This study describes the hypotensive, cardiosuppressant and vasodilator activities of Hyoscyamus niger crude extract (Hn.Cr). Hn.Cr, which tested positive for alkaloids, coumarins, flavonoids, sterols, tannins and terpenes, caused a dose-dependent (10-100 mg/kg) fall in the arterial blood pressure (BP) of rats under anesthesia. In guinea-pig atria, Hn.Cr exhibited a cardiodepressant effect on the rate and force of spontaneous atrial contractions. In isolated rabbit aorta, Hn.Cr (0.01-1.0 mg/ml) relaxed the phenylephrine (PE, 1 microM) and K(+) (80 mM)-induced contractions and suppressed PE (1 microM) control peaks obtained in Ca(++)-free medium similar to that caused by verapamil. The vasodilator effect of Hn.Cr was endothelium-independent as it was not opposed by N (omega)-nitro-L-arginine methyl ester in endothelium-intact rat aortic preparations and also occurred at a similar concentration in endothelium-denuded tissues. These data indicate that Hyoscyamus niger lowers BP through a Ca(++)-antagonist mechanism.

Pharmacological studies on hypotensive, diuretic and vasodilator activities of chrysin glucoside from Calycotome villosa in rats.

The present study was undertaken in normotensive anaesthetized male rats that received a continuous perfusion of a chrysin glucoside isolated from the flowers and leaves of Calycotome villosa subsp intermedia at a dose of 2.5 mg/kg, or furosemide (control diuretic) at a dose of 0.5 mg/kg. Compared with the control rats receiving NaCl (0.9%), the urine flow, glomerular filtration and electrolyte excretion (Na+, K+) increased significantly in rats treated with chrysin glucoside (p < 0.001). A similar effect was observed in the rats perfused with furosemide. Intravenous injections of bolus doses (1-3 mg/kg) of the chrysin glucoside to anaesthetized rats elicited an immediate and dose-dependent decrease in mean arterial blood pressure (MABP). Pretreatment of the rats with the nitric oxide synthase inhibitor, l-NOArg (10 mg/kg), reduced partially, but significantly (p < 0.01), the maximal decrease in MABP elicited by chrysin glucoside. In the rat isolated aorta preparation, chrysin glucoside (10-100 microm) inhibited in a concentration-dependent manner the noradrenaline (1 microm) induced contractions (IC(50) = 52 microm). This relaxant activity of chrysin glucoside was significantly reduced by incubation of the endothelium-intact rings with l-NOArg (100 microm), (80 +/- 4.7% vs 48 +/- 5.06% in the absence of L-NOArg). In conclusion, these results demonstrate a diuretic and hypotensive action of a chrysin glucoside from Calycotome villosa in anaesthetized rats and indicating an action on renal function, and an active vascular relaxation mediated partially through nitric oxide release.

Antinociceptive and antipyretic activities of the Zizyphus oxyphylla Edgew. leaves.

A methanol extract of Zizyphus oxyphylla Edgew leaves has been investigated for its analgesic and antipyretic activities in Adult Wistar and Swiss albino mice of either sex at 50, 100 and 200 mg/kg orally. The extract demonstrated marked antipyretic activity against Brewer's yeast-induced pyrexia in rats. The extract demonstrated significant peripheral analgesic effect in the acetic acid-induced writhing test in mice. The phytochemical tests revealed that the extract contained alkaloids, anthraquinones, flavonoids, glycosides, phenols, resins, saponins and tannins using standard procedures. In conclusion, the present study suggests that the methanol extract of Zizyphus oxyphylla Edgew leaves possesses potent antipyretic and antinociceptive activities and thus validates its use in the treatment of pain and fever.

Cardio-selective inhibitory effect of the betel nut extract: possible explanation.

Chewing of betel nut, the seed of Areca catechu, is associated with a host of physical and psychological effects while it is also traditionally used in constipation and hypertension. In this study, we report the cardio-selective cholinomimetic activity of the betel nut crude extract (Ac.Cr). Ac.Cr, that tested positive for saponins, tannins, phenols, alkaloids and terpenes, exhibited dose-dependent atropine-sensitive inhibition of isolated guinea-pig atrial contractility with an EC50 value of 0.93 microg/ml (0.57-1.51, 95% CI). In rabbit jejunum, Ac.Cr showed atropine-sensitive spasmogenicity with an EC50 of 7.31 microg/ml (5.41-9.88, 95% CI) showing that it is around 8 times more potent in the cardiac than the intestinal preparation. Both carbachol and physostigmine exhibited acetylcholine-like stimulant activity in jejunum with the latter being more potent in jejunum than in atrial tissues. Activity-directed fractionation of Ac.Cr yielded fractions with similar cholinergic activity in atria and jejunum except the aqueous fraction being 6 times more potent in the atria. Arecoline, the known betel nut compound with cholinergic activity showed similar potency in both tissues while catechin and tannic acid exhibited intestinal spasmolytic effect but were inactive in atria. The results show the cardio-selective inhibitory effect of Ac.Cr which might possibly be due to selective gut-spasmolytic behaviour of catechin and tannic acid thus reducing the cholinomimetic activity of Ac.Cr in the gut though the preferential binding of the constituents of betel nut extract at muscarinic receptor subtypes in heart cannot be ignored.

Mechanisms of the anti-hypertensive effect of Hibiscus sabdariffa L. calyces.

Previous studies have demonstrated the anti-hypertensive effects of Hibiscus sabdariffa L. (HS) in both humans and experimental animals. To explore the mechanisms of the anti-hypertensive effect of the HS, we examined the effects of a crude methanolic extract of the calyces of HS (HSE) on vascular reactivity in isolated aortas from spontaneously hypertensive rats. HSE relaxed, concentration-dependently, KCl (high K(+), 80 mM)- and phenylephrine (PE, 1 microM)-pre-contracted aortic rings, with a greater potency against the alpha(1)-adrenergic receptor agonist. The relaxant effect of HSE was partly dependent on the presence of a functional endothelium as the action was significantly reduced in endothelium-denuded aortic rings. Pretreatment with atropine (1 microM), L-NAME (10 microM) or methylene blue (10 microM), but not indomethacin (10 microM), significantly blocked the relaxant effects of HSE. Endothelium-dependent and -independent relaxations induced by acetylcholine and sodium nitroprusside, respectively, were significantly enhanced in aortic rings pretreated with HSE when compared to those observed in control aortic rings. The present results demonstrated that HSE has a vasodilator effect in the isolated aortic rings of hypertensive rats. These effects are probably mediated through the endothelium-derived nitric oxide-cGMP-relaxant pathway and inhibition of calcium (Ca(2+))-influx into vascular smooth muscle cells. The present data further supports previous in vivo findings and the traditional use of HS as an anti-hypertensive agent.

Antagonism of antinociceptive effect of hydro-ethanolic extract of Hypericum perforatum Linn. by a non selective opioid receptor antagonist, naloxone.

Hydro-ethanolic crude extract of Hypericum perforatum Linn. family hypericaceae (St. John's Wort) aerial parts (Hp. Cr) was studied for its possible antinociceptive effect against acetic acid-induced abdominal constriction assay in mice. Hp. Cr (10-20 mg kg(-1)), opium (10-30 mg kg(-1)), morphine (0.75-3.0 mg kg(-1)) and aspirin (50-100 mg kg(-1)) showed dose-dependent antinociceptive effect. In animals treated with naloxone (0.5 mg kg(-1)), the antinociceptive effect of Hp. Cr was significantly reduced similar to that of opium, while effect of aspirin remained unchanged. These results suggest that the antinociceptive effect of Hypericum perforatum may be mediated through activation of opioid receptors.

Presence of calcium antagonist activity explains the use of Syzygium samarangense in diarrhoea.

Syzygium samarangense (Family; Myrtaceae) or 'makopa', as it is commonly known, is native to Malaysia, some islands of Indonesia and to Far East in general. This study was undertaken to rationalize the use of this plant in hypermotility states of the gut. The hexane extract of S. samarangense (Ss.Hex) was found to dose-dependently (10-3000 microg/mL) relax the spontaneously contracting isolated rabbit jejunum. When tested for a possible calcium channel blocking (CCB) activity, the extract (10-1000 microg/mL) relaxed the high K+-induced contractions and also decreased the Ca++ dose-response curves in a dose-dependent manner (30-100 microg/mL), confirming the CCB activity. Four flavonoids isolated from the hexane extract were tested for a possible spasmolytic activity. All flavonoids, identified as: 2'-hydroxy-4',6'-dimethoxy-3'-methylchalcone (SS1), 2',4'-dihydroxy-6'-methoxy-3',5'-dimethylchalcone (SS2), 2',4'-dihydroxy-6'-methoxy-3'-methylchalcone (SS3) and 7-hydroxy-5-methoxy-6,8-dimethylflavanone (SS4), showed dose-dependent (10-1000 microg/mL) spasmolytic activity with SS2 being the most potent. These results indicate that the presence of compounds with spasmolytic and calcium antagonist activity may be responsible for the medicinal use of the plant in diarrhoea.

Studies on the antihypertensive, antispasmodic, bronchodilator and hepatoprotective activities of the Carum copticum seed extract.

This study describes the antihypertensive, antispasmodic, bronchodilator and hepatoprotective activities of the aqueous-methanolic extract of Carum copticum Benth. seeds (CSE) to rationalize some of its traditional uses. CSE (3-100 mg/kg) caused a dose-dependent fall in arterial blood pressure in anaesthetized rats. In isolated rabbit aorta and jejunum preparations, CSE (0.1-3.0 mg/ml) caused an inhibitory effect on the K+-induced contractions. The calcium channel blocking (CCB) effect was confirmed when CSE shifted the Ca2+ dose-response curves (DRCs) to right similar to verapamil. In isolated guinea-pig tracheal preparations, it caused inhibition of carbachol and K+-induced bronchoconstriction at 0.1-1.0 mg/ml as well as shifted the dose-response curves (DRCs) of carbachol and histamine to the right with suppression of maximum response suggestive of non-specific bronchodilator effect mediated possibly through CCB. Pretreatment of rats with CSE (500 mg/kg orally for 2 days at 12 h intervals) prevented paracetamol (640 mg/kg) and CCl4 (150 ml/kg)-induced rise in serum alkaline phosphatase (ALP) and aminotransferases (AST and ALT). The same dose of CSE was able to prevent the CCl4-induced prolongation in pentobarbital-induced sleeping time in mice confirming its hepatoprotectivity. These results indicate the presence of calcium antagonist(s) in Carum copticum seeds and thus provides sound mechanistic basis for some of their folkloric uses.

Studies on the protective effects of caffeic acid and quercetin on chemical-induced hepatotoxicity in rodents.

Caffeic acid and quercetin, the well-known phenolic compounds widely present in the plant kingdom, were investigated for their possible protective effects against paracetamol and CCl4-induced hepatic damage. Paracetamol at the oral dose of 1 g/kg produced 100% mortality in mice while pretreatment of separate groups of animals with caffeic acid (6 mg/kg) and quercetin (10 mg/kg) reduced the death rate to 20% and 30%, respectively. Oral administration of sub-lethal dose of paracetamol (640 mg/kg) produced liver damage in rats as manifested by the significant (P<0.01) rise in serum levels of aminotransferases (aspartate transaminase (AST) and alanine transaminase (ALT)) compared to respective control values. The serum enzyme values were significantly (P<0.01) lowered on pretreatment of rats with either caffeic acid (6 mg/kg) or quercetin (10 mg/kg). Similarly, the hepatotoxic dose of CCl4 (1.5 ml/kg; orally) also raised significantly (P<0.05) the serum AST and ALT levels as compared to control values. The same dose of the caffeic acid and quercetin was able to prevent CCl4-induced rise in serum enzymes. Caffeic acid and quercetin also prevented the CCl4-induced prolongation in pentobarbital sleeping time confirming their hepatoprotectivity. These results indicate that caffeic acid and quercetin exhibited hepatoprotective activity possibly through multiple mechanisms.

Cholinesterase inhibitory and spasmolytic potential of steroidal alkaloids.

A new steroidal alkaloid, isosarcodine (1) along with four known bases, sarcorine (2), sarcodine (3), sarcocine (4) and alkaloid-C (5) were isolated from the MeOH extract of Sarcococca saligna. The structures of these alkaloids were identified by spectral data interpretation. These compounds were subjected to acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibition studies, and were found to be noncompetitive inhibitors of AChE (Ki = 21.8, 90.3, 32.2, 16.0 and 50.0 microM, respectively) and uncompetitive or noncompetitive inhibitors of BChE (Ki = 8.3, 7.5, 15.6, 5.0 and 12.0 microM, respectively). The compounds (2-5) also showed dose-dependent spasmolytic activity in the rabbit jejunum intestinal preparations and also relaxed the high K+ (80 mM)-induced contraction, indicative of a calcium channel-blocking mechanism. Structure-activity relationship suggested that the nitrogen substituents at C-3 and/or C-20 of steroidal skeleton and the hydrophobic properties of the pregnane skeleton are the key structural features contributed to the inhibitory potency of these steroidal alkaloids against AChE and BChE.

The in vitro effect of aqueous extract of Nigella sativa seeds on nitric oxide production.

The in vitro effect of aqueous extract of Nigella sativa seeds on nitric oxide (NO) production by murine macrophages was studied. Murine peritoneal macrophages were pre-incubated with the extract and then activated with Escherichia coli lipopolysaccharride. NO production was measured after 24 hours by spectrophotometry. The plant extract caused a dose-dependent decrease in NO production. Dialyzed preparation of the extract did not affect NO production. However, the boiled fraction of the extract resulted in a dose-dependent inhibition of NO apparently comparable to that of the whole extract. These results indicate that the aqueous extract of N. sativa seeds exhibits an inhibitory effect on nitric oxide production by murine macrophages and the active component(s) is/are non-protein in nature. In view of the fact that nitric oxide is a pro-inflammatory mediator, this study validates the traditional use of the Nigella sativa seeds for the treatment of rheumatism.

Nutrient digestibility of broiler feeds containing different levels of variously processed rice bran stored for different periods.

Nutrient digestibility of broiler feeds containing different levels of variously processed rice bran stored for varying periods was determined. A total of 444 Hubbard male chicks were used to conduct four trials. Each trial was carried out on 111 chicks to determine digestibility of 36 different feeds. Chicks of 5 wk age were fed feeds containing raw, roasted, and extruded rice bran treated with antioxidant, Bianox Dry (0, 125, 250 g/ton), stored for a periods of 0, 4, 8, and 12 mo and used at levels of 0, 10, 20, and 30% in feeds. Digestibility coefficients for fat and fiber of feeds were determined. Increasing storage periods of rice bran significantly reduced the fat digestibility of feed, whereas no difference in fiber digestibility was observed. Processing of rice bran by extrusion cooking significantly increased digestibility of fat even used at higher levels in broiler feeds. Interaction of storage, processing, and levels was significant for fat digestibility. Treatments of rice bran by different levels of antioxidant had no effect on digestibility of fat and fiber when incorporated in broiler feed.