Prolonged Oral Administration Research Articles

Novel quinazoline and acetamide derivatives as safe anti-ulcerogenic agent and anti-ulcerative colitis activity

Two novel quinazoline derivatives named as; 3-[(4-hydroxy-3-methoxy-benzylidene)-amino]-2-p-tolyl-3H-quinazolin-4-one (5) and 2-p-Tolyl-3-[3,4,5-trimethoxy-benzylidene-amino]-3H-quinazolin-4-one (6) in addition to one acetamide derivative named as 2-(2-Hydroxycarbonylphenylamino)-N-(4-aminosulphonylphenyl) 11 were synthesized, and evaluated for their anti-ulcerogenic & Anti-Ulcerative colitis activities.All of the three compounds showed curative activity against acetic acid induced ulcer model at a dose of 50mg/kg, they produced 65%, 85% & 57.74% curative ratio for compounds 5, 6 &11 respectively. The effect of the tested compounds 5, 6 &11 at dose 50 mg/kg were significantly (P<0.01) more effective than dexamesathone (0.1 mg/kg) in reducing all parameters.Compounds showed curative activity of for peptic ulcer (induced by absolute alcohol (at a dose of 50 mg/kg, it produced Curative of control ulcer 56.00%, 61.70% & 87.1% for compounds 5, 6 &11 respectively at dose 50 mg/kg, while the standard drug (Omeprazole 20 mg/kg) produced 33.3%. In both tests, the activity of our target compounds were higher than the standard drugs used for treatment of peptic ulcer and ulcerative colitis. No side effects were reported on liver and kidney functions upon prolonged oral administration of this compounds.

Oral administration of essential oils and main components: Study on honey bee survival and Nosema ceranae development

Diverse parasites and pathogens affect productivity and survival of honey bees. Plant secondary metabolites are potential alternative treatments, however, their effect has been little studied on microsporidian diseases. Furthermore, there is poor information about the toxicity resulting from prolonged oral administration of these substances to bees. In this research, we evaluated in vivo effects of different essential oils and main components (MCs) on bee survival and Nosema ceranae development under an ad libitum non-choice regimen. Substances administered on sucrose syrup to newly emerged bees at different concentrations were avidly consumed and caused different survival performances. Nevertheless, sublethal doses of substances did not control the parasite.

Acute and Sub-chronic Oral Toxicity Evaluation of Eucalyptus globulus Essential Oil-Water Emulsion in Mice

Background: plants are composed of bioactive chemicals some of which may be toxic. Therefore, scientists advocate for toxicological studies to be carried out in order to ensure the safety of drugs. Accordingly, this study was aimed to assess the possible toxic effects of Eucalyptus glubulus essential oil- water emulsion after oral administration in mice. Methods: The essential oil used in this study was obtained by extraction of the fresh leaves of E. glubulus through hydro-distillation technique. In the acute toxicity study, the mice placed in the seven treatment groups were administered with randomly selected 0.5 ml/kg initial dose up to 3.5 ml/kg body weight spaced by 0.5 ml/kg. The control animals received vehicle. All animals were gavaged the designated dose once and observed for the next two weeks. In the sub-chronic study, a control and two treatment groups, each containing twelve animals were used. Based on the findings of acute toxicity study, the two treatment groups were received 1.5 and 2.0 ml/kg body weight test doses respectively for three months in 24 hours interval. At the end study period, blood samples were taken by cardiac puncture and then, all animals were sacrificed and the vital organs were collected and processed for histopathological examination. Results: In the acute toxicity study, some toxicity signs and deaths of mice were recorded only at doses higher than 2.0 ml/kg body weight. 1 out of 6 mice in a group treated with 3.0 ml/kg and, 4 out of 6 animals in a group treated with 3.5 ml/kg body weight test doses of the emulsion were died following the administration. Therefore, the oral route LD50 of the emulsion falls between 3 ml/kg and 3.5 ml/kg. In the sub-chronic treatment, no death was recorded, and there was no statistically significant (p>0.05) change in the body weight, evaluated hematological and biochemical parameters of blood. However, minor pathological changes like; pyknosis and vacuolations of hepatocytes in some liver sections, and perivascular lymphocytic infiltrations and hyaline casts in renal tubules of some kidney sections, were observed in mice treated with 2.0 ml/kg body weight test dose. Conclusions and recommendations: The emulsion at relatively lower doses does not produce obvious toxic effects after acute and prolonged oral administration in mice. However, further investigation is needed to confirm this.

Macrophage-Mediated Clofazimine Sequestration Is Accompanied by a Shift in Host Energy Metabolism

Prolonged (8 weeks) oral administration of clofazimine results in a profound pharmacodynamic response—bioaccumulation in macrophages (including Kupffer cells) as intracellular crystal-like drug inclusions (CLDIs) with an associated increase in interleukin-1 receptor antagonist production. Notably, CLDI formation in Kupffer cells concomitantly occurs with the formation of macrophage-centric granulomas. Accordingly, we sought to understand the impact of these events on host metabolism using 1H-nuclear magnetic resonance metabolomics. Mice received a clofazimine or vehicle-enriched (sham) diet for at least 8 weeks. At 2 weeks, the antimicrobial activity of clofazimine was evident by changes in urine metabolites. From 2 to 8 weeks, there was a striking change in metabolite levels indicative of a reorientation of host energy metabolism paralleling the onset of CLDI and granuloma formation. This was evidenced by a progressive reduction in urine levels of metabolites involved in one-carbon metabolism with corresponding increases in whole blood, and changes in metabolites associated with lipid, nucleotide and amino acid metabolism, and glycolysis. Although clofazimine-fed mice ate more, they gained less weight than control mice. Together, these results indicate that macrophage sequestration of clofazimine as CLDIs and granuloma formation is accompanied by a profound metabolic disruption in energy homeostasis and one-carbon metabolism.

Alibertia edulis (L.C. Rich.) A.C. Rich – A potent diuretic arising from Brazilian indigenous species

Ethnopharmacological relevanceAlthough Alibertia edulis (L.C. Rich.) A.C. Rich decoction is used in Brazilian folk medicine due to its possible antihypertensive effect, this species has never been critically investigated as a hypotensive drug. So, the aim of this study was to evaluate the possible hypotensive and antihypertensive effects of the oral administration of Alibertia edulis aqueous extract (AEAE) in normotensive and hypertensive rats, and evaluate its inter-relation with a possible diuretic activity. Material and methodsDifferent doses of AEAE (20, 65 and 200mg/kg) were tested on the mean arterial pressure (MAP) of normotensive Wistar rats and after induction of renovascular hypertension (two-kidney, one-clip Goldblatt model). In addition, the diuretic effects of AEAE were compared with hydrochlorothiazide (HCTZ) in an acute and repeated-dose treatment for 7 days. Volume, sodium, potassium, chloride, calcium contents, pH and density were estimated in urine samples collected after 8 or 24h. Plasma sodium, potassium, total protein, urea, creatinine, AST and ALT concentrations were measured in samples collected at the end of the experimental period (seventh day). Finally, the antioxidant activity of the AEAE was assessed using the DPPH radical scavenging and ferric ions reducing power assay. ResultsThe intraduodenal administration of the HCTZ and AEAE significantly reduced, in a dose-dependent manner, the MAP in both normotensive and hypertensive rats. Otherwise, the heart rate was not affected by any treatment. Acute and prolonged oral administration of AEAE (200mg/kg) and HCTZ caused a significant increase in volume and urinary concentrations of sodium, potassium and chloride. Moreover, urinary calcium concentration was significantly increased after administration of AEAE (200mg/kg). Finally, AEAE was able to present important in vitro antioxidant properties. ConclusionThe results obtained have shown that AEAE presents potent diuretic activity and significant hypotensive and antihypertensive effect. In addition, this study may confirm part of the pharmacological activity popularly attributed to this species and opens perspective for the future use in various renal and cardiovascular diseases.

The morphological changes in the internal organs of laboratory animals after prolonged oral administration of gold nanoparticles

Recently gold nanoparticles (GNPs) have been actively studied as photothermal converters, drug carriers, and imaging agents in a wide range of applications in cancer diagnosis and therapy. The prolonged peroral administration of GNPs in a range of sizes was performed to investigate the morphological changes and their reversibility in the internal organs of laboratory animals. In this study, GNPs with average diameters of 2[Formula: see text]nm, 15[Formula: see text]nm and 50[Formula: see text]nm were administered during 30 days, and the reversibility of morphological changes was investigated 14 days after administration. After the prolonged administration of GNPs, the severity of morphological changes in the liver, kidney, spleen and lymph nodes depended on the nanoparticle size. Specifically, 50[Formula: see text]nm nanoparticles caused the most pronounced dystrophic and necrobiotic effects, whereas the smallest 2[Formula: see text]nm nanoparticles caused proliferative changes. Most importantly, the development of pathological processes was reversible, as evidenced by the gradual restoration of the organ structure at 14 days after the end of GNPs administration.

Injectable long acting chitosan/tripolyphosphate microspheres for the intra-articular delivery of lornoxicam: Optimization and in vivo evaluation

Chitosan microspheres were formulated for the intra-articular delivery of lornoxicam in knee osteoarthritis, to minimize associated side-effects after prolonged oral administration. Ionotropic-gelation technique was employed using tripolyphosphate as anionic cross-linker. Full-factorial design experiment was conducted to optimize lornoxicam entrapment-efficiency%. Formulations were assessed for their particle size, in-vitro drug release, Scanning electron microscopy, Differential-scanning-calorimetry and Fourier transform infra-red spectroscopy studies. Changing independent variables, chitosan pH, TPP pH and lornoxicam concentration resulted in different values of entrapment-efficiency% ranging from 13.5%±0.35 to 59.5%±2.2. Particle size ranged from 3.57μm±0.02 to 6.12μm±0.00 and lornoxicam%release was prolonged for up to 8days. SEM results showed spherical shape of the microspheres. FTIR and DSC studies confirmed the crosslinking of chitosan with tripolyphosphate. In-vivo therapeutic effect of lornoxicam microspheres was investigated using Monosodiumiodoacetate (MIA) induced osteoarthritis model in rats. Optimized formula showed long-term in-vivo anti-inflammatory effect relative to lornoxicam solution injected intra-articularly with significant reduction of histological, inflammatory and biochemical parameters of osteoarthritis.

Prolonged Oral Administration of a Pan-Retinoic Acid Receptor Antagonist Inhibits Spermatogenesis in Mice With a Rapid Recovery and Changes in the Expression of Influx and Efflux Transporters.

We have previously shown that oral administration of a pan-retinoic acid receptor antagonist in mice daily at 2.5 mg/kg for 4 weeks reversibly inhibited spermatogenesis, with no detectable side effects. To elucidate the lowest dose and the longest dosing regimen that inhibits spermatogenesis but results in complete restoration of fertility upon cessation of administration of the drug, we examined the effects of daily doses as low as 1.0 mg/kg with dosing periods of 4, 8, and 16 weeks. We observed 100% sterility in all regimens, with restoration of fertility upon cessation of the drug treatment even for as long as 16 weeks. There was no change in testosterone levels in these males and the progeny examined from 2 of the recovered males were healthy and fertile, with normal testicular weight and testicular histology. Strikingly, a more rapid recovery, as assessed by mating studies, was observed at the lower dose and longer dosing periods. Insight into possible mechanisms underlying this rapid recovery was obtained at 2 levels. First, histological examination revealed that spermatogenesis was not as severely disrupted at the lower dose and with the longer treatment regimens. Second, gene expression analysis revealed that the more rapid recovery may involve the interplay of ATP-binding cassette efflux and solute carrier influx transporters in the testes.

Adverse effects of long-term administration of fluvoxamine on haematology, blood biochemistry and fertility in male albino rats: a possible effect of cessation.

Fluvoxamine is recommended as first-line treatment for a number of obsessive-compulsive disorders, anxiety disorders, social phobia, and post-traumatic stress disorder and panic disorder. The adverse effects of prolonged oral administration of fluvoxamine on haematology, biochemical parameters and fertility in male rats were evaluated in this study. Sixty adult male rats were allocated into 5 equal groups and orally treated with fluvoxamine 9mgkg-1 b.wt. (low therapeutic dose, LTD) and 27mgkg-1 b.wt. (high therapeutic dose, HTD), while the control rats received 0.5ml distilled water for a period of 8weeks. The 4th and 5th groups were gavaged with LTD and HTD of fluvoxamine for 8weeks and then left untreated for another 8weeks (recovery groups). HTD of fluvoxamine induced leukocytosis, lymphocytosis and monocytosis. LTD and HTD of fluvoxamine evoked hepatic, renal and cardiac dysfunction. Moreover, fluvoxamine treatment might lead to the risk of male infertility, which is indicated by its deleterious impacts on spermiogram and steroidogenesis hormones. They also induced oxidative stress, apoptosis in testicular tissue. Fortunately, the previous alterations were mostly reversed during the recovery period.

CC-486 (Oral Azacitidine) Monotherapy in Patients with Acute Myeloid Leukemia (AML)

Background: When administered subcutaneously (SC), the epigenetic modifier, azacitidine (AZA) is shown to improve overall survival in patients (pts) with AML compared with conventional care regimens (Fenaux, JCO, 2010; Dombret, Blood, 2015). CC-486, the oral formulation of AZA, is in clinical development for use in hematologic malignancies. An early dose-finding study of CC-486 administered on a 7-day schedule showed it to be safe and clinically active in pts with AML (Garcia-Manero, JCO, 2011; Gore, Blood, 2011). Extending CC-486 dosing for more than 7 days may increase AZA exposure to leukemic cells over the treatment (Tx) cycle to improve response.Objective: Evaluate the safety and efficacy of CC-486 administered in extended dosing schedules (14 days or 21 days per 28-day cycle) in pts with AML.Methods: Pts with AML from two phase I/II studies were sequentially assigned to receive CC-486 in 1 of 4 extended dosing regimens: 300mg QD x14 days (d)/28d (14-QD), 300mg QD x21d/28d (21-QD), 200mg BID x14d/28d (14-BID), or 200mg BID x21d/28d (21-BID). Hematologic responses (complete remission [CR], partial remission [PR], CR with incomplete hematologic recovery [CRi], marrow CR [mCR]), were assessed using IWG criteria for AML (Cheson, J Clin Oncol, 2003), and hematologic improvement (HI) and transfusion independence (TI) were assessed using IWG criteria for MDS (Cheson, Blood, 2006). Tx-emergent adverse events (TEAEs) were graded by NCI-CTCAE v3.1 or v4.0.Results: In all, 23 pts with AML participated in the studies: 14-QD n=4, 21-QD n=12, 14-BID n=4, 21-BID n=3. Median age was 68 years (range 44 - 93), 48% were male, 22% and 78% had ECOG performance status scores of 0-1 or 2, respectively, median % bone marrow blasts was 25%, and median baseline hematology counts were: Hgb 9.1 g/dL (7.4 - 11.7), platelets 33 x 109/L (3 - 435), and ANC 0.3 x 109/L (0 - 6.4). Median time from diagnosis was 1.1 month (range -0.2 - 82.1). Eleven pts (52%) had prior MDS and 13 pts (57%) had received prior Tx for MDS (n=5) or AML (n=8), including 5 pts (38%) who had received hypomethylating agents (HMAs). The median number of CC-486 Tx cycles for all pts was 4 (1 - 9) (Figure). Overall response (CR, PR, CRi, mCR, HI or TI) rate was 48% (11/23), with generally comparable responses and response rates across the 4 dosing regimens (Table 1). Responses were observed in 4 pts (50%) who were relapsed/refractory to prior AML Tx. Two pts (40%) who had received prior HMA Tx responded, 1 of whom attained CR. HI was achieved by 32% (7/22) of pts, and 20% (3/15) of RBC transfusion-dependent pts attained RBC TI. Six pts with prior MDS (55%) responded during Tx. One pt proceeded to transplant. Febrile neutropenia (30%) and anemia (22%) were the most common grade 3-4 TEAEs (Table 2). Incidences of grade 3-4 TEAEs were generally similar across the 4 CC-486 dosing regimens.Conclusions: CC-486 administered in extended 14- or 21-day per cycle schedules was well tolerated. One-half of these older pts with AML had a hematologic response, including pts who had prior MDS, or AML that failed or lost response to prior Tx. Interestingly, pts who previously received injectable HMA Tx had hematologic responses (including CR) with CC-486, likely due to the alternative methylation pattern obtained with prolonged oral administration (Laille, Plos One, in press). Ease of administration of CC-486 can facilitate long-term Tx for sustained disease modification, provide an additional option for failure of IV/SQ HMA therapy, and may improve outcomes in combination Tx regimens, which warrant investigation. [Display omitted] [Display omitted] [Display omitted] DisclosuresSavona:Karyopharm: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Celgene: Membership on an entity's Board of Directors or advisory committees; Gilead: Membership on an entity's Board of Directors or advisory committees; Incyte: Membership on an entity's Board of Directors or advisory committees, Research Funding. Gore:Celgene: Consultancy, Honoraria, Research Funding. Kolibaba:Janssen: Research Funding; Acerta: Research Funding; Amgen: Research Funding; Celgene: Research Funding; Gilead: Consultancy, Research Funding; Pharmacyclics: Research Funding; TG Therapeutics: Research Funding; Cell therapeutics: Research Funding; GSK: Research Funding; Novartis: Research Funding; Genentech, Inc.: Research Funding; Seattle Genetics: Research Funding. Ukrainskyj:Celgene Corporation: Employment, Equity Ownership. Kumar:Celgene Corporation: Employment, Equity Ownership. Dong:Celgene Corporation: Employment, Equity Ownership. Hetzer:Celgene Corporation: Employment, Equity Ownership. Skikne:Celgene Corporation: Employment, Equity Ownership.

Longitudinal study of effects of oral dosage of Bifidobacterium bifidum G9-1 on Japanese cedar pollen-induced allergic nasal symptoms in guinea pigs.

Previous studies using experimental animal models have reported the beneficial effects of probiotics on allergic responses; however, their long-term effects on allergic nasal symptoms in clinical settings have not yet been elucidated in detail. In the present study, a guinea pig allergic rhinitis model involving repeated inhalation challenges with a natural allergen, Japanese cedar pollen, was used to examine the longitudinal effects of Bifidobacterium bifidum G9-1 (BBG9-1) on allergic nasal symptoms. BBG9-1 was administered orally once a day. Amelioration of nasal blockage was consistently observed throughout the experimental period in the BBG9-1-treated group. Although challenge-induced sneezing was not significantly inhibited in the BBG9-1-treated group, prolonged treatment with BBG9-1 slightly reduced the frequency of sneezing. Antigen-specific IgE antibody production was also not inhibited in the BBG9-1-treated group. Increases in the numbers of eosinophils and neutrophils in nasal cavity lavage fluid collected after pollen challenge were almost completely suppressed by BBG9-1 treatment, whereas those in mast cell mediators, histamine and cysteinyl leukotrienes were not. In contrast, increases in the levels of nitric oxide metabolites were potently suppressed. Furthermore, prolonged BBG9-1 treatment markedly suppressed exogenous leukotriene D4 -induced nasal blockage. Thus, prolonged oral administration of BBG9-1 suppresses Japanese cedar pollen-induced allergic nasal symptoms. The inhibitory mechanisms responsible may involve reductions in the responsiveness of target organs, such as endothelial cells in nasal mucosal blood vessels, to chemical mediators.

Abstract 2599: Discovery and pharmacological characterization of the second generation of Btk inhibitors with improved target selectivity and enhanced in vivo efficacy

Abstract Bruton's tyrosine kinase (Btk) is an essential component of the B-cell receptor (BCR) signaling pathways regulating survival, activation, proliferation, and differentiation of B lymphocytes. The first Btk inhibitor, Ibrutinib, has demonstrated a significant clinical efficacy in a variety of B-cell malignancies. Through structure-guided approach, we have discovered two series of novel and potent Btk inhibitors, represented by EBI-1266 and EBI-1367, respectively. Both of these compounds showed high potency in inhibiting Btk kinase activity and growth of a number of aggressive lymphoma cell lines driven by aberrant BCR signaling, with IC50 value within low nanomolar range. Mechanistic studies examining BCR signaling pathway in related B cell lymphoma cell lines revealed that both EBI-1266 and EBI-1367 potently inhibited Btk phosphorylation and downstream Erk phosphorylation, similar to Ibrutinib. Furthermore, each compound showed unique and significant superiority over Ibrutinib: EBI-1266 had a cleaner selectivity profile against a panel of kinases with a cysteine residue in the conserved catalytic domain, and EBI-1367 exhibited &amp;gt;10-fold higher in vivo exposure in multiple preclinical species. In a tumor xenograft mouse model where tumor growth is dependent on Btk activity, both EBI-1266 and EBI-1367 showed significant oral anti-tumor activities, with EBI-1367 being more efficacious than Ibrutinib, consistent with a higher exposure of EBI-1367 in both blood and tumor tissues. No evidence of overt toxicities was observed in rodents with prolonged oral administration for two weeks with doses at least 3-fold above a highly efficacious dose. In summary, the pharmacological profiles of EBI-1266 and EBI-1367 indicate that these compounds have a great potential to become the next generation of Btk inhibitors, offering advantages in pharmaceutical development, enhanced in vivo efficacy and reduced toxicities. We are currently performing IND-enabling studies aiming to initiate phase I clinical trials for these Btk inhibitors. Citation Format: Jiayin Zhang, Dong Liu, Ru Shen, Yinfa Yan, Liuqing Yang, Minsheng Zhang, Jun Feng, Beibei Fu, Jerry Hu, Biao Lu, Hong Wan, Lei Zhang, Weikang Tao, Lianshan Zhang, Jingsong Cao. Discovery and pharmacological characterization of the second generation of Btk inhibitors with improved target selectivity and enhanced in vivo efficacy. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2599. doi:10.1158/1538-7445.AM2015-2599

Damage and regeneration of small intestinal enterocytes under the influence of carrageenan induces chronic enteritis

The food additive carrageenan (E407) is a sulfated polysaccharide extracted from red seaweeds which has been widely used in the food industry as a thickener and gelling agent. Several varying models of carrageenan-induced acute inflammation have been developed to allow study of the anti-inflammatory activity of drugs and to evaluate the role of inflammatory mediators (Nantel et al. 1999). Carrageenan has been used to induce both acute and chronic inflammation in laboratory animals, including peritonitis (de Carvalho et al. 2013), pleurisy (Renard et al. 2014) and arthritis (Arun et al. 2013). Oral consumption of carrageenan has been shown to result in chronic intestinal inflammation and neoplasia in intestines of experimental animals (Tobacman 2001). However, the pathogenesis of carrageenan-induced inflammation is still poorly understood. In particular, levels of apoptotic activity in small intestine after prolonged oral administration of carrageenan are unknown. This study examines morphological features of small intestine and evaluates apoptotic biochemical indices in homogenate of small intestine of rats after prolonged oral administration of carrageenan.

Prolonged oral administration of Gastrodia elata extract improves spatial learning and memory of scopolamine-treated rats.

Gastrodia elata (GE) is traditionally used for treatment of various disorders including neurodegenerative diseases such as Alzheimer's disease. To investigate the neuroprotective effect of GE, amyloid-β peptide (Aβ)-treated PC12 cells were cultured with GE aqueous extract. In vitro assay demonstrated that 50 µM of pre-aggregated Aβ was lethal to about a half portion of PC12 cells and that Aβ aggregate-induced cell death was significantly decreased with GE treatment at ≤10 mg/mL in a dose-dependent manner. To further examine in vivo cognitive-improving effects, an artificial amnesic animal model, scopolamine-injected Sprague-Dawley rats, were orally administered the extract for 6 weeks followed by behavioral tests (the passive avoidance test and Morris water maze test). The results showed that an acute treatment with scopolamine (1 mg/kg of body weight) effectively induced memory impairment in normal rats and that the learning and memory capability of scopolamine-treated rats improved after prolonged administration of GE extract (50, 250 and 500 mg/kg of body weight for 6 weeks). These findings suggest that a GE regimen may potentially ameliorate learning and memory deficits and/or cognitive impairments caused by neuronal cell death.

Role of naringenin in protection against diabetic hyperalgesia and tactile allodynia in male Wistar rats.

Hyperalgesia and allodynia are among the common manifestations of painful diabetic neuropathy. Naringenin (NA) has some biological activities, including anti-inflammatory, analgesic, and antidiabetic effects. We investigated the effects of NA administration at different doses, 20, 50, and 100 mg/kg, on streptozotocin (STZ)-induced hyperalgesia and allodynia in rats. The animals received saline or NA (20, 50, and 100 mg/kg, p.o.; once daily) for 8 weeks. Hyperalgesia was assessed by tail flick (TF) and formalin tests. Von Frey filaments were used for tactile allodynia evaluation. At the end, all rats were weighed and underwent plasma glucose and superoxide dismutase measurement. Diabetes caused significant hyperalgesia and allodynia during the above tests. NA 50 and 100 mg/kg reversed chemical and thermal hyperalgesia in diabetic rats. There were no significant differences in pain responses between NA (50 and 100 mg/kg)-treated diabetic rats and pregabalin-treated diabetic animals. Administration of NA 20 mg/kg did not alter pain-related behaviors in control and diabetic groups compared to the respective control ones. NA 50 and 100 mg/kg restored hyperglycemia as well as the decreased levels of (superoxide dismutase) SOD activity in diabetic rats. The body weight of treated diabetic rats increased significantly compared to untreated diabetics. Prolonged oral administration of NA (50 and 100 mg/kg) ameliorated some aspects of diabetic neuropathy by causing hypoglycemia and increasing the levels of antioxidant enzyme SOD. Therefore, NA makes a good candidate for treatment of diabetic neuropathy in clinical studies.

A new way to produce hyperketonemia: Use of ketone ester in a case of Alzheimer's disease

BackgroundProviding ketone bodies to the brain can bypass metabolic blocks to glucose utilization and improve function in energy-starved neurons. For this, plasma ketones must be elevated well above the ≤0.2 mM default concentrations normally prevalent. Limitations of dietary methods currently used to produce therapeutic hyperketonemia have stimulated the search for better approaches. MethodDescribed herein is a new way to produce therapeutic hyperketonemia, entailing prolonged oral administration of a potent ketogenic agent—ketone monoester (KME)—to a patient with Alzheimer's disease dementia and a pretreatment Mini-Mental State Examination score of 12. ResultsThe patient improved markedly in mood, affect, self-care, and cognitive and daily activity performance. The KME was well tolerated throughout the 20-month treatment period. Cognitive performance tracked plasma β-hydroxybutyrate concentrations, with noticeable improvements in conversation and interaction at the higher levels, compared with predose levels. ConclusionKME-induced hyperketonemia is robust, convenient, and safe, and the ester can be taken as an oral supplement without changing the habitual diet.

Evaluation of the antioxidant activity of Marjoram plant (Origanium Majorana L.) in CCl 4-intoxicated rats.

Aim: This work was performed to evaluate the antioxidant activity and hepatoprotective effect of methanolic extract of Marjoram plant (Origanium Majorana L.) in CCl4- intoxicated rats. Materials and Methods: A total of 25 mature albino rats (Sprague Dawley) from both sexes, weighting from140 to 170 g were used. The animals were randomly divided into 5 groups each of 5 animals. These groups were used to investigate the in vivo hepatoprotective and antioxidant effects of the tested plant extract when given orally in two doses (375 and 750 mg/kg b.wt.). The first group was kept as normal healthy control. The other four groups were given CCl4 (1 ml/kg b.wt. s.c) for five days. One of these was used as a CCl4-intoxicated non treated control. Group III and IV were given the methanol extract of Origanium majorana plant orally at doses of 375 and 750 mg/kg b.wt. per day, for two months. Group V was used as a standard group and received silymarin orally at a dose of 100 mg/kg b.w. for two months. The antioxidant activity of methanol extract of Origanium majorana plant was studied in rats liver homogenate after the prolonged oral administration (2 months) via determination of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) enzyme activity. The in vitro antioxidant activity was compared to DPPH and measured spectrophotometrically. The activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) in the different groups was evaluated as well as the histopathological examination of the liver. Results: Oral administration of Origanium majorana plant increased the antioxidant enzymes and significantly (P < 0.05) decreased liver enzyme levels when given in repeated doses. Conclusion: Thus methanolic extract of Origanium majorana possesses a significant antioxidant activity.

Phenolic Profiling and Evaluation of Contraceptive Effect of the Ethanolic Extract of Salsola imbricata Forssk. in Male Albino Rats.

Reported researches dealing with either composition or bioactivity of Salsola imbricata are limited. This study was conducted aiming to investigate the phenolic composition of the plant and evaluate its efficacy as male contraceptive. Polyphenols, namely, phenolic acids and flavonoids, were qualitatively and quantitatively analysed by RP-HPLC in the hydrolysed methanol extract using two different wavelengths, 280 and 330 nm. The efficiency of different solvents in extracting the plant phenolics was assessed via spectrophotometric determination of the total phenolic and flavonoid contents. Acute toxicity study was carried out on the ethanolic extract to ascertain its safety prior to biological evaluation. The contraceptive effect was assessed, in male rats, by oral administration of the extract at two doses (250 and 500 mg/kg b. wt.), over a period of 65 days. HPLC analyses allowed the identification and quantification of a total of 13 and 8 components in the hydrolysed-methanol extract; the overall phenolic composition was dominated by quercitrin (12.692%) followed by coumaric acid (4.251%). Prolonged oral administration of the ethanolic extract caused slight reduction in the testis weight only. A significant decrease in the sperm count was observed (P < 0.01) in the two treated groups while significant decrease in the epididymal sperm motility was only observed in the high dose group. Morphological abnormalities were observed in sperms of treated animals. No distinct change in serum FSH, LH, and testosterone concentration was recorded. The histopathological findings supported to a high extent these results. The male contraceptive activity of Salsola imbricata could be ascribed to its phenolic components, especially quercitrin.

Effects of prolonged oral administration of dicyandiamide to dairy heifers on excretion in urine and efficacy in soil

Oral administration of the nitrification inhibitor dicyandiamide (DCD) to grazing ruminants for excretion in urine represents a targeted mitigation strategy to reduce nitrogen (N) losses from grazed pastures. A field trial and allied laboratory incubation study were conducted to examine the effects of oral administration of DCD to non-lactating Friesian dairy heifers on excretion of DCD in urine and efficacy in soil. Dairy heifers were orally administered DCD daily at three treatment levels (low, medium and high; 12, 24 and 36gDCDheifer−1day−1, respectively) and compared to a nil-DCD control group over a 90-day continuous dosing period. There were no adverse effects of DCD administration on heifer health or growth, as inferred by live-weight gain and measured blood metabolite levels. Prolonged administration of DCD to dairy heifers resulted in the sustained excretion of DCD in the urine over 90 days and inhibition of nitrification of urinary-N in the silty peat soil for up to 56 days (incubated at 20°C; P<0.001). Field soil sampling (0–75mm depth) of individual urine patches for DCD analysis revealed that a 3-fold increase in the rate of DCD administered resulted in a similar increase in the concentration of DCD voided in the urine and subsequently deposited in urine patches (median equivalent DCD application rates of 22, 36 and 59kgha−1 for the low, medium and high DCD treatment levels, respectively; P<0.001). However, large differences (up to 40-fold) existed between individual urine patches in the rate of DCD deposited at each treatment level, which showed a positively skewed distribution. This study highlights the viability of prolonged daily administration of DCD to ruminants for sustained excretion in urine and effective inhibition of nitrification in soil as a practical targeted mitigation technology to reduce urinary-N losses from grazed pastures.

Metabolite profiling of polyphenols in a Terminalia chebula Retzius ayurvedic decoction and evaluation of its chemopreventive activity

Ethnopharmacological relevanceThe decoction of Terminalia chebula fruit is an ayurvedic remedy whose prolonged oral administration is prized as a generic intestinal and hepatic detoxifying agent. Its administration is suggested also under the perspective of a reduced risk of cancer, metabolic and cardiovascular diseases. Aim of the studyTo evaluate the phytochemical profile and the chemopreventive potential of Terminalia chebula fruit decoction prepared according to the ayurvedic decoction recipe. Materials and methodsThe quali- and quantitative metabolite profiling of polyphenols was obtained using HPLC–UV/DAD and HPLC-ESI-MS. The crude decoction and purified compounds were tested for their capability to interact with the EphA2-ephrin-A1 system and for their antimutagenic properties against dietary and environmental mutagens (AA, 2-NF, NaN3, and heterocyclic amines IQ, MeIQ, MeIQx, Glu-P1, Glu-P2,) in the Ames–Salmonella/microsome assay, with and without enzymatic induction. ResultsThe decoction was found to contain 3,4,6-tri-O-galloyl-d-glucose (55.87mg/g), chebulic acid (54.03mg/g), β-punicalagin (41.25mg/g), corilagin (40.31mg/g), α-punicalagin (35.55mg/g), chebulagic acid (29.09mg/g), gallic acid (27.96mg/g) 1,3,4,6-tri-O-galloyl-β-d-glucose (24.25mg/g) chebulinic acid (20.23mg/g), 1,2,3,4,6-penta-O-galloyl-d-glucose (13.53mg/g), ellagic acid (8.00mg/g), 1,6-di-O-galloyl-d-glucose (4.16mg/g). An inhibitory effect was recorded in both Salmonella typhimurium TA98 and TA100 strains against the mutagenic activity of heterocyclic amines (22–61%), promutagen AA (91–97%) and directly acting mutagen 2-NF (52%) with but not against NaN3 (7%). Galloyl derivatives allowed an inhibition of mutagenicity induced by MeIQ above 80% at 0.01mol/plate. Both decoction and purified compounds were able to modulate the EphA2-ephrinA1 system, suggesting a potential multiple chemopreventive mechanism. ConclusionsThe traditional ayurvedic decoction of Terminalia chebula may harbour a potential as a safe and low-cost chemopreventive agent at the intestinal level, if administered according to the ayurvedic specifications. Moreover, its recourse may enhance the presence of some polyphenolic constituents.