Significant Dose-dependent Increase Research Articles

Neuropharmacological assessment of the methanolic stem back extract of Anopyxis klaineana (Pierre) Engl. (Rhizophoraceae) in mice

BackgroundMedicinal plants have recently attracted attention on a global scale for therapeutic interventions in the field of neuroscience. Anopyxis klaineana, a medicinal plant widely used in West Africa, has been studied extensively for its anti-inflammatory effects. However, studies on its neuropharmacological effects are quite limited. This study therefore assessed the neuropharmacological potential of the methanolic stem bark extract of Anopyxis klaineana (EAK) in murine models. Materials and methodsMice were treated orally with a methanolic stem back extract of Anopyxis klaineana (EAK) at 10, 30 or 100 mg/kg followed by testing in the elevated plus-maze (EPM) and open field (OF) tests for anxiolytic-like effects. EAK's antidepressant-like potential was also evaluated in the tail suspension test (TST), while anticonvulsant and central analgesic effects were evaluated with the pentylenetetrazol (PTZ)-induced convulsion threshold and tail withdrawal tests respectively at 30, 100 or 300 mg/kg. The extract was additionally subjected to chromatographic analyses using high resolution HPLC-MS. ResultsTotal ion current chromatogram of EAK revealed several peaks suggestive of secondary metabolites of the plant extract. EAK significantly (p<0.05) reduced depression-like behaviours in the TST although not as potent as fluoxetine (ED50: EAK=43.39±2.5; fluoxetine=1.16±1.6) (F3,20=9.663, p = 0.0004). It also showed significant anticonvulsive effect by reducing the frequency and duration of PTZ-induced convulsions (ED50=48.53±2.5 mg/kg) (F3,19=5.136, p = 0.0091). This anticonvulsant effect was seen in both clonic and tonic convulsions at 30 and 300 mg/kg. Additionally, EAK-treated mice showed higher latency to tail withdrawal in the analgesia test. We also observed significant (p<0.05) dose-dependent increase in anxiolytic-like effects in EAK-treated animals in the EPM (F3,20=10.77, p = 0.0002) and OF tests (F3,22=5.079, p = 0.008). ConclusionsThe methanolic Anopyxis klaineana stem back extract exhibits anxiolytic-like, antidepressant-like, analgesic and anticonvulsant effects in murine models for assessing neuropharmacological activity and is a good source for novel neuroactive compounds.

Acetaldehyde induces similar cytotoxic and genotoxic risks in BEAS-2B cells and HHSteCs: involvement of differential regulation of MAPK/ERK and PI3K/AKT pathways.

Long-term use of alcohol and cigarettes is associated with millions of deaths each year, directly or indirectly. The carcinogen acetaldehyde is both a metabolite of alcohol and the most abundant carbonyl compound in cigarette smoke, and co-exposure of them is usual and primarily leads to liver and lung injury, respectively. However, few studies have explored the synchronic risk of acetaldehyde on the liver and lung. Here, we investigated the toxic effects and related mechanisms of acetaldehyde based on normal hepatocytes and lung cells. The results showed that acetaldehyde caused significant dose-dependent increases of cytotoxicity, ROS level, DNA adduct level, DNA single/double-strand breakage, and chromosomal damage in BEAS-2B cells and HHSteCs, with similar effects at the same doses. The gene and protein expression and phosphorylation of p38MAPK, ERK, PI3K, and AKT, key proteins of MAPK/ERK and PI3K/AKT pathways regulating cell survival and tumorigenesis, were significantly upregulated on BEAS-2B cells, while only protein expression and phosphorylation of ERK were upregulated significantly, the other three decreased in HHSteCs. When either the inhibitor of the four key proteins was co-treated with acetaldehyde, cell viabilities were almost unchanged in BEAS-2B cells and HHSteCs. Thus, acetaldehyde could synchronically induce similar toxic effects in BEAS-2B cells and HHSteCs, and MAPK/ERK and PI3K/AKT pathways seem to be involved in different regulatory mechanisms.

Investigating the effects of 7-ketocholesterol on retinal pigment epithelium bioenergetics.

Age-related macular degeneration (AMD) is associated with formation of drusen, clusters of lipids, and oxidized lipid products under the retinal pigment epithelium (RPE). 7-Ketocholesterol (7KC) is a form of oxidized cholesterol present in drusen and is hypothesized to play a role in AMD pathogenesis. The association of 7KC with cellular toxicity and inflammation, key elements of AMD pathology, has been demonstrated. However, the effects of 7KC on altering RPE bioenergetics, a potentially important pathologic process in AMD, are unclear. Herein, we describe the effects of non-lethal doses of 7KC on the bioenergetics and phenotype of RPE cells in culture. Metabolic analysis demonstrated a significant dose-dependent increase in total ATP production rates that was driven primarily by an increase in glycolysis. The increase in glycolysis was accompanied by an increase in glucose uptake and increased expression of hexokinase 1. Increased levels of Translocase of Outer Mitochondrial Membrane 20and NADH:Ubiquinone Oxidoreductase Core Subunit S1, Succinate dehydrogenase, Ubiquinol-Cytochrome C Reductase Core Protein 2, Cytochrome C Oxidase II, and ATP synthase subunit beta, proteins involved in oxidative phosphorylation (OXPHOS), were also seen. However, specific electron transport chain activity remained unchanged. 7KC-treated cells also demonstrated a change in cellular morphology with decreased expression of epithelial markers. In summary, 7KC has significant effects on the bioenergetics and morphology of RPE cells reflective of findings seen in clinical AMD.

Olanzapine’s Cytogenetic Effect on T Lymphocytes in Systemic Lupus Erythematosus and Rheumatoid Arthritis Patients: In Vitro Study

This study will investigate olanzapine's cytogenetic behavior in cultured human T lymphocytes in patients with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). Three olanzapine solutions were added in cultures of peripheral blood lymphocytes of healthy individuals, SLE, and RA patients. After 72 hours of incubation, the cultured lymphocytes were plated on glass slides and stained with the fluorescence plus Giemsa method. Sister chromatid exchanges (SCEs), proliferation rate index (PRI), and mitotic index (MI) were measured with the optical microscope. There was a statistically significant (p=0.001) dose-dependent increase of SCEs in SLE and RA patients compared to healthy individuals and a statistically significant (p=0.001) reduction of PRI and MI in the highest concentration in the SLE group. Moreover, Spearman's rank correlation coefficient was applied to calculate the correlation between SCEs, PRI, and MI. Negative significant correlations were noticed for both patient groups concerning SCEs-PRI alterations and SCEs-MI alterations. Conversely, positive correlations were noticed for both patient groups for PRI-MI alterations. Conclusions: Olanzapine affects T lymphocytes from SLE and RA patients by modifying DNA replication procedures and DNA damage response. Considering the use of olanzapine in neuropsychiatric symptoms of SLE, further in vivo studies are necessary to evaluate its effect on human DNA.

Abstract 2890: Investigating dynamic IDO-Kyn-Ahr pathway induced tumor immunosuppression using imaging modality to optimize therapeutic Intervention

Abstract Despite great clinical responses in several cancers to immune checkpoint blockade (ICB) intervention, there persist many mechanisms of tumor evasion. Tryptophan catabolism by enzymes Indoleamine-2,3-dioxygenase 1 (IDO1) and tryptophan-2,3-dioxygenase 2 (TDO2) into metabolite L-Kynurenine (Kyn) has been shown to drive an immunosuppressive tumor microenvironment (TME). We have shown tumors overexpressing IDO1 recruit and activate the Treg/macrophage suppressive axis and blockade of aryl hydrocarbon receptor (AHR) activation abrogates this. Preclinical models demonstrate inhibition of IDO1 delayed tumor growth and increased T cell function. However, a phase III clinical trial in melanoma patients combining IDO1 inhibition with anti-PD-1 blockade showed no improved efficacy when compared to anti-PD-1 blockade. Kyn can act as a key signaling molecule through activation of AHR. We hypothesize that AHR engagement by kyn within the TME is highly dynamic and elevated AHR engagement must coincide with intervention to maximize therapeutic response. Using an AHR biosensor, we designed a construct with a dioxin response element (DRE) promoter that is engaged upon AHR-ligand complex, translocated to the nucleus, and drives a luciferase gene with a luminescence readout. This allows us to use an image-guided approach to track AHR bioactivity in real-time. In-vitro, we were able to validate our construct using transduced murine B16 melanoma and 4T1 breast cancer models. We treated transduced cells with endogenous AHR ligands, kynurenine, and AHR agonist (FICZ), and observed a significant dose-dependent increase in AHR reporter readout. Additionally, the AHR inhibitor (CH-223191) abrogated AHR activity and subsequent luminescence below control threshold. We implanted both DRE transduced B16 (B16-DRE) and 4T1 (4T1-DRE) intradermally and intramammary fat pad respectively. Tumors were imaged and we observed total luminescence flux corresponding to increased tumor growth. After three weeks, we assessed IDO, AHR, and HIF1a expression which all converge on the IDO-Kyn-AhR pathway using a broad panel of lymphocyte and myeloid markers from tumors and spleen. We identified high expression of IDO, AHR, and HIF1a markers in myeloid cells with an increased frequency of dendritic cells and macrophages. Future experiments will focus on using our reporter construct in the context of ICB treatment and assessing AHR activity overtime as well as sampling peripheral blood to assess the magnitude of tryptophan, kyn, and other polycyclic hydrocarbons including gut microbiome-derived indoles. The characterization of the IDO-Kyn-AhR modulation after therapeutic intervention will inform the rational design of treatment regimens to abrogate immune suppression associated with AHR activity in combination with ICB treatment. Citation Format: Mamadou Alpha Bah, Inna Serganova, Sadna Budhu, Tanya Schild, Jonathan F. Khan, Kayvan R. Keshari, Jedd D. Wolchok, Taha Merghoub. Investigating dynamic IDO-Kyn-Ahr pathway induced tumor immunosuppression using imaging modality to optimize therapeutic Intervention [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2890.

Viloxazine Increases Cortical Serotonin Without Inhibiting Serotonin Reuptake at Doses Used to Treat ADHD

AbstractBackgroundMost FDA-approved ADHD treatments increase norepinephrine (NE) and dopamine (DA); however, our prior preclinical studies of the non-stimulant ADHD treatment viloxazine ER (Qelbree®) demonstrated that viloxazine also increases serotonin (5-HT). A prior microdialysis study showed increases in NE, DA, and 5-HT in the rat prefrontal cortex (PFC); however, the 50 mg/kg dose resulted in supratherapeutic plasma concentrations. Viloxazine is a moderate affinity selective NE reuptake inhibitor, structurally different than traditional SSRI antidepressants. Viloxazine has negligible activity at the serotonin reuptake transporter (SERT), suggesting viloxazine has a different mechanism of 5-HT PFC elevation than SSRIs. The current microdialysis study was undertaken to further characterize if viloxazine affects 5-HT and its 5-HIAA metabolite at therapeutically relevant plasma concentrations. Results are compared to similar microdialysis studies of SSRIs.MethodsRats were implanted with I-shaped microdialysis probes connected to a microperfusion pump, delivering artificial cerebrospinal fluid, in the PFC. After a 2-hour baseline period, viloxazine (1, 3, 10, or 30 mg/kg) was administered (ip). Dialysate samples were collected from the interstitial fluid (ISF) of the PFC before and after dosing. LC-MS/MS was used to determine the dialysate concentrations of viloxazine and viloxazine-induced changes in NE, 5-HT, and their respective metabolites, DHPG and 5-HIAA. Viloxazine plasma concentrations were also measured.Animal research was approved by the Institutional Animal Care and Use Committee and conducted in accordance with the National Research Council’s Guide for the Care and Use of Laboratory Animals.ResultsViloxazine administration resulted in significant dose-dependent increases in ISF NE levels and corresponding decreases in DHPG (NE metabolite) at all doses tested, reflecting viloxazine’s activity as a NET inhibitor. Viloxazine treatment also resulted in a dose-dependent elevation of ISF 5-HT levels in the PFC. Of the doses tested, 30 mg/kg was found to be clinically relevant as it induced ISF concentrations approximating unbound plasma concentrations in pediatric ADHD patients. At this dose, 5-HT levels were significantly increased over baseline and higher than vehicle levels. Coincident changes in 5-HIAA concentrations were not observed, reaffirming viloxazine’s lack of activity as a SERT inhibitor.ConclusionViloxazine induced dose-dependent increases in NE and 5-HT in the PFC, a critical target region for ADHD therapies. At clinically relevant viloxazine plasma concentrations, 5-HT was increased in the PFC. Unlike SSRIs, which correspondingly decrease the 5-HT metabolite in the PFC (indicating serotonin reuptake inhibition), viloxazine did not affect 5-HIAA levels. Thus, viloxazine increases cortical 5-HT levels by a different mechanism than SSRIs. Whether 5-HT effects aid in viloxazine therapeutic efficacy in ADHD is yet unknown.FundingSupernus Pharmaceuticals, Inc.

Immunostimulatory and Antimicrobial Claims of Super-7 Herbal Mixture (SHM) and Odogwu Cleanser Herbal Mixture (OCHM) Commercially Available in Enugu State, Nigeria

The continuous increase in resistance associated with conventional drugs is gradually shifting focus to herbal remedies as an alternative globally. This study was designed to investigate the immunostimulatory and antimicrobial claims of Super-7 herbal mixture (SHM) and Odogwu cleanser herbal mixture (OCHM) commercially available in Enugu State, Nigeria. Immunomodulatory properties of the products on the Delayed Type Hypersensitivity reaction (DTHr), the Primary and Secondary humoral response and the <i>in vivo</i> leucocyte mobilization were evaluated using a total of one hundred and forty-six (146) Swiss albino rats. The spectrum of antimicrobial activity was evaluated by agar well diffusion method using eight (8) selected microorganisms. The LD₅₀ of both products was greater than 100% ml/kg body weight. OCHM elicited significant (p<0.05) dose dependent increase in total leucocyte and neutrophil counts compared to Levamisole (2.5mg/kg) and the untreated group. A 100% ml/kg SHM (78.07%) also exhibited significantly (p<0.05) higher oedema inhibitory potentials compared to the standard drug (levamisole = 65.78%). Samples of SHM demonstrated dose dependent antibacterial activities against <i>B. subtilis </i>and <i>S. typhi</i>, while only exhibiting antifungal activity against<i> C. albicans </i>(IZD = 9.0±0.0 mm). OCHM was observed to inhibit <i>E. coli, S. typhi, S. aureus, B. subtilis, S. typhi </i>and the fungus<i> C. albicans</i>. An average bioload of 2.8 x 10⁴±1.0 CFU/ml which was higher than the average count (2.8 x 10⁴±1.0 CFU/ml) obtained for OCHM. Conclusively, this study showed that samples of SHM and OCHM exhibited significant immunostimulatory and antimicrobial properties, but also recorded the presence of microbial contaminants.

Maximising performance and phosphorus utilisation of warm water fish through phytase supplementation

Phosphorus (P) is a critical mineral nutrient for fish but its availability in plant-based aquafeeds is usually low. To avoid its over-supplementation and the consequent increase of feed cost, excretion and, ultimately, environmental pollution, phytase supplementation has been used as a strategy to improve the bio-availability of P present in plant-based feeds as well as overall nutrient digestibility. This study assessed the efficacy of a new generation bioengineered microbial phytase (HiPhorius™) on the growth performance, whole-body nutrient retention, nutrient digestibility, and P excretion of Nile tilapia (Oreochromis niloticus), European seabass (Dicentrarchus labrax), and gilthead seabream (Sparus aurata). In the trial, the fish were fed diets enriched with plant-protein rich diets. Five dietary treatments were tested over 94 days: a control diet (CTRL), three diets supplemented with phytase at graded doses (500, 1000, and 2000 FTY/kg feed), and a diet supplemented with either dicalcium or monocalcium phosphate (DCP/MCP). Results showed that feeds supplemented with the phytase led to a significant dose-dependent increase of final body weight, specific growth rate, and protein efficiency ratio (FBW, SGR, PER), and a significant reduction of FCR, in comparison to the CTRL treatment. Supplementation with the phytase also notably reduced P excretion down to 23.37 ± 1.69, 28.46 ± 4.07, and 18.86 ± 1.05 mg/kg/day of P excretion for Nile tilapia, European seabass, and gilthead seabream, respectively, as compared to the treatment with inorganic P supplementation (118.82 ± 3.87, 127.81 ± 2.58, and 117.09 ± 8.15 mg/kg/day of P excretion for Nile tilapia, European seabass, and gilthead seabream, respectively). Overall, results show that the phytase tested here is an effective strategy to contribute to the economic and environmental sustainability of aquaculture production in different fish species.

Spermatogenic and antiradical activities of the hydro-ethanolic extract of Strychnos camptoneura (longaniaceae) trunk barks in Wistar rats

The objective of the present study were to evaluate the spermatogenic and antiradical potentialities of the hydro-ethanolic extract of Strychnos camptoneura trunk bark in male wistar rats. For this purpose, 20 rats were divided into 4 lots of 5 animals and received respectively for lots 1 and 2 distilled water and testosterone enanthate. Lots 3 and 4 were treated with the hydro-ethanolic extract of Strychnos camptoneura (100 and 250 mg/kg). The results obtained show that, the administration of hydro-ethanolic extract of Strychnos camptoneura (100 and 250 mg /kg/po) orally in rats caused significant dose-dependent increases (p &lt; 0.05; p &lt; 0.01 and p &lt; 0 .001) in sperm concentration and motility rates compared to rats of the control group treated with distilled water. On the other hand, the vitality rates of sperm and normal sperm were only significant (p &lt; 0.01) at the respective doses of 100 and 250 mg /kg of Strychnos camptoneura compared to the negative controls. In addition, there was no variation in sperm pH (p ˃ 0.05) at any dose level. Chemical analysis of the hydro-ethanol extract of Strychnos camptoneura revealed the existence of phenolic compounds especially polyphenols and total flavonoids endowed with antiradical and antioxidant properties. The improvement of the characteristics of the semen and the reduction of the DPPH radical observed in the present work, justify its biological potentialities and its use in traditional medicine in the treatment of male infertility.

Influence of Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis, and prebiotic inulin on dysbiotic dental biofilm composition ex vivo.

Probiotic bacterial supplementation has shown promising results in the treatment of periodontitis and the maintenance of periodontal health. The purpose of this investigation was to evaluate the influence of Lactobacillus reuteri or Bifidobacterium animalis subsp. lactis supplementation with and without prebiotic inulin on biofilm composition using an ex vivo biofilm model. Subgingival plaque specimens from three periodontitis-affected human donors were used to grow biofilms on hydroxyapatite disks in media supplemented with varying combinations of prebiotic inulin, Lactobacillus reuteri, and Bifidobacterium animalis subsp. lactis. Relative abundances of bacterial genera present in mature biofilms were evaluated using 16S rRNA next-generation sequencing. Diversity metrics of microbial communities were evaluated using a next-generation microbiome bioinformatics platform. Inulin supplementation produced statistically significant dose-dependent increases in relative abundances of Lactobacillus and Bifidobacterium species (p<0.001) with concomitant decreases in relative abundances of Streptococcus, Veillonella, Fusobacterium, Parvimonas, and Prevotella species (p<0.001). Inoculation with L. reuteri or B. animalis subsp. lactis increased the relative abundance of only the supplemented probiotic genera (p<0.05). Supplemental inulin led to a statistically significant decrease in biofilm alpha diversity (p<0.001). The described ex vivo model appears suitable for investigating the effects of probiotic bacteria, prebiotic oligosaccharides, and combinations thereof on biofilm composition and complexity. Within the limitations imposed by this model, results from the present study underscore the potential for prebiotic inulin to modify biofilm composition favorably. Additional research further elucidating biologic rationale and controlled clinical research defining therapeutic benefits is warranted.

The Antioxidant, Antibacterial and Anti-Biofilm Properties of Rapeseed Creamed Honey Enriched with Selected Plant Superfoods.

The aim of the study is to evaluate the effect of the addition of selected fruits and herbs belonging to the "superfoods" category for the bioactivity of a rapeseed honey matrix. Flavored creamed honeys with nine types of various additives (2 and 4% of content) were prepared and analyzed for the content of total phenols, flavonoids, antioxidant (FRAP, DPPH and ABTS) and antibacterial activity against four strains of bacteria. Additionally, the impact of three months of storage on the antioxidant properties of the products obtained was examined. The significant dose-dependent increase in the content of bioactive ingredients and antioxidant capacity in spiced honeys, as compared to control honey, was observed. The highest enrichment was obtained for the addition of powdered sea buckthorn leaves and black raspberry fruits. Honey with the addition of sea buckthorn leaves inhibited the growth of P. aeruginosa, S. aureus and K. pneumonia, whereas honeys with black raspberry and blackcurrant fruits showed activity only on the latter two strains. Furthermore, what is more interesting, honey supplemented with sea buckthorn leaf and black raspberry fruits inhibited S. aureus biofilm formation at the sub-minimum inhibitory concentrations (sub-MICs), showing a dose-dependent anti-biofilm effect.

Effects of a New Kappa Agonist (Fluorophenyl Derivative of Imidazo[1,2-&lt;i&gt;а&lt;/i&gt;]benzimidazole) on the Rat Genome

Selective kappa-opioid receptor (KOR) agonists are considered a promising group of substances for developing opioid analgesics characterised with an original mechanism of action without the risk of respiratory depression and drug addiction. Previous studies identified a fluorophenyl derivative of imidazo[1,2-a]benzimidazole (RU-1205) with a KOR-based mechanism of analgesic action established in in vitro and in vivo experiments.The aim of the study was to assess the effect of 9-(2-morpholinoethyl)-2-(4-fluorophenyl)imidazo[1,2-a]benzimidazole dihydrochloride on the level of DNA damage in rats after a single subcutaneous injection.Materials and methods. The study was conducted in adult white outbred laboratory rats of both sexes. DNA damage was estimated using the comet assay. The study involved a single subcutaneous injection of an aqueous solution of RU-1205 in three doses: 1, 10, and 100 mg/kg. The authors used intraperitoneal methyl methanesulfonate (40 mg per kg of animal body weight) as a positive control and 0.9% NaCl (100 μL per 100 g of animal body weight) as a negative control.Results. A single subcutaneous injection of RU-1205 to rats did not produce a significant dose-dependent increase in % tail DNA when compared with the state of the corresponding organ/tissue cell genome in negative control animals after normal saline administration at the same time points. In the negative control groups, % tail DNA in cells of various organs/tissues ranged from 1.83% to 3.82% (median values [25–75%]). On the contrary, the administration of 40 mg/kg of genotoxic methyl methanesulfonate led to an increase in damaged DNA in all studied organs and tissues when compared with negative control animals.Conclusions. The study of 9-(2-morpholinoethyl)-2-(4-fluorophenyl)imidazo[1,2-a] benzimidazole dihydrochloride genotoxicity demonstrated that a single subcutaneous injection of 1, 10, or 100 mg/kg of RU-1205 to rats did not damage the cell genome of the studied organs.

The intestinal permeability marker FITC-dextran 4kDa should be dosed according to lean body mass in obese mice

AimsTo investigate the influence of the dose in the FITC-Dextran 4kDa (FD-4) permeability test in an obese mouse model, we tested the bodyweight dose regimen and a lean body mass-based dose regimen in high fat diet (HFD) mice and low fat diet (LFD) mice. We hypothesized that the FD-4 permeation result would be dose-dependent.MethodsThe two dose regimens were compared in HFD and LFD mice. Furthermore, we conducted a dose-response study to test the effect of a low or high dose of FD-4 in weight-stratified lean mice. Gene analysis of tight junctions was also carried out.ResultsThe FD-4 intestinal permeability test was dose-dependent as we found a significant increase in plasma levels of FD-4 in obese mice with the bodyweight dose regimen. However, this difference was not detectable with the lean body mass dose regimen, even with variability-adjusted group sizes. However, the qPCR analysis revealed a decrease in tight junction gene expression in obese mice. Furthermore, we found a dose-dependent significant increase in FD-4 measured in plasma samples in lean mice. No significant difference in intestinal weight was observed between lean and obese mice.ConclusionEvaluation of the intestinal permeability by FD-4 with the typical bodyweight dose regimen in obese mice will be confounded by the significant difference in dose given when compared to a lean control group. If the test dose is based on lean body mass, no significant difference in intestinal permeability is observed, even with large group sizes. Furthermore, we showed a dose-dependent difference in plasma FD-4 levels in lean mice. Therefore, we conclude that the dose should be based on lean body mass for the FD-4 permeability test if mice with considerable obesity differences are to be compared or to use another test with fixed doses.

Evaluation of Antinociceptive Activity of Ficus Religiosa Root Extract in Swiss Albino Mice

Background &#x0D; Ficus religiosa, commonly known as peepal, is widely distributed in Indian subcontinent. It has been used as an antiepileptic, aphrodisiac, analgesic, anti-inflammatory and laxative in traditional medicine.&#x0D; Objective To explore the analgesic effect of aqueous root extract of Ficus religiosa using thermal and chemical models of pain in swiss albino mice.&#x0D; Method The aqueous aerial root extract of Ficus religiosa was prepared using soxhlet apparatus. The anti-nociceptive effect of the extract at doses of 50 and 100 mg/kg was evaluated using peripheral (acetic acid-induced abdominal writhing), spinal (tail flick) and supra-spinal (hot plate) behavioral models of pain. All data were presented as Mean ± SEM. Statistical differences between Ficus religiosa (50 and 100 mg/kg) and standard control groups were evaluated using Mann-Whitney U test.&#x0D; Result There was significant dose dependent increase in the mean reaction time compared to the vehicle control in hot plate and tail- flick test. In acetic acid induced writhing test, mice treated with Ficus religiosa (50 and 100 mg/kg) exhibited significant dose-dependent decrease in the mean number of writhes (57.45% and 79.20% respectively) compared to the vehicle control. The activity of Ficus religiosa extract at doses of 50 and 100 mg/kg was equipotent to Standard control (Morphine and Indomethacin) used in different test models.&#x0D; Conclusion The extract of Ficus religiosa possesses both central and peripheral analgesic activity thus validating the traditional use of this plant in the management of pain.

Protective role of Withania somnifera root extract on lipid peroxidation of Erythrocytes

Withania Somnifera (Ashwagandha) is an important medicinal plant that has been used in Ayurvedic and indigenous medicine. It is also known as Indian ginseng. It exerts antioxidant, anti-inflammatory and immune modulatory activities. In the present study, we studied the free radical scavenging activity of Withania Somnifera root extract. The radical scavenging effect of the extract was studied by its effect on hydroxy radicals generated by Fe-ascorbate-H2O2 system. Our results showed that Withania Somnifera root extract (1-10,000 µg/ ml) resulted in a significant dose dependent increase in percent inhibition of hydroxylation. Further, we studied the in vitro effect of Withania Somnifera root extract on Lipid peroxidation in terms of MDA level of human erythrocytes. Our results showed a significant dose dependent decrease in lipid peroxidation at a concentration of (1-1000 µg/ ml) as evident by decrease in level of MDA. Maximum effect was observed at 100-1000 µg/ ml. Withania Somnifera root extract thus can act as a potential antioxidant containing compound and protects erythrocytes from oxidative damage by either directly scavenging the free radical, sparing it or activating other enzymatic processes. At higher concentration (10,000 µg/ ml), Withania Somnifera extract increased the oxidative stress as evident from erythrocyte MDA level which became significantly more than the maximum effect shown at 1000 µg/ ml of extract.

Effects of Two Months Durational Vitamin E Therapy on Paraquat-inflicted Liver Damage in Wistar Rats (Rattus norvegicus)

Paraquat is a highly poisonous chemical that is commonly used as a herbicide (plant killer) to control weeds and grasses. It has been found to disrupt the levels and activities of some liver enzyme parameters. This experimental study was aimed at assessing the effect of Vitamin E in restoring liver physiology in paraquat-induced liver poisoning in rats. In this study, 200 male rats with a mean weight of 0.20.02kg, with 50 rats in each group of four, labeled A, B, C, and D. The "A" group received no paraquat, while the "B", "C", and "D" groups received 0.02g, 0.04g, and 0.06g of paraquat per kilogram of rat every two weeks for three months respectively. Each group also had a subgroup treated only with vitamin E and the Vit. E + paraquat subgroup BVE, CVE, and DVE but AVE was treated with paraquat but only Vit E. The blood was taken and the liver function was assayed. The result showed that there was a significant dose-dependent increase (P-value&lt;0.05) in the levels liver enzymes in the subgroups treated with paraquat alone (A0, B0, C0 and D0). After treatment with vit E, the result revealed that there a significant decrease (P-value&lt;0.05) in the level of the liver enzymes when compared to the corresponding subgroups without Vit. E treatment. This study has shown that vit E can ameliorate the effect of paraquat liver poisoning particularly when the treatment is administered weekly for two months.

Acute and sub-lethal toxicity in African mud catfish (Clarias gariepinus, Burchell, 1822) exposed to some pesticides

Pesticides from agricultural run-off pose a severe threat to non-target organisms, such as fishes. This study was carried out to evaluate acute toxicity and histological and genotoxic effects (erythrocytic nuclear abnormalities) of lethal and sub-lethal concentrations of three commonly used pesticides: Atrazine, Butachlor and Glyphosate on the African mud catfish (Clarias gariepinus). Fishes were exposed to the pesticides for 96h periods to determine their LC50 and the sub-lethal effect at various concentrations (1/10th, 1/100th , 1/1000th 96h LC50) over 28 days. The 96h LC50 values were 7.63mg/l, 0.7mg/l and 15.97mg/l for atrazine, butachlor and glyphosate, respectively. Histological sections of the liver of C. gariepinus exposed to the three pesticides showed mild vascular congestion but no necrosis in the tissue. There was a significant (p&lt;0.5) dose-dependent increase in micronuclei and nuclear abnormalities in the erythrocytes of exposed C. gariepinus compared to the control by 28 days. The study confirmed that C. gariepinus are at risk of adverse effects from exposure to pesticides. Discharge of agricultural run-off around water bodies should be prevented or prohibited to avoid adverse effects on aquatic life.

Structurally diverse fentanyl analogs yield differential locomotor activities in mice

Synthetic narcotics have been implicated as the single greatest contributor to increases in opioid-related fatalities in recent years. This study evaluated the effects of nine fentanyl-related substances that have emerged in the recreational drug marketplace, and for which there are no existing or only limited in vivo data. Adult male Swiss Webster mice were administered fentanyl-related substances and their effects on locomotion as compared to MOR agonist standards were recorded. In locomotor activity tests, morphine (100, 180 mg/kg), buprenorphine (1, 10 mg/kg), fentanyl (1, 10 mg/kg), cyclopropylfentanyl (1, 10 mg/kg), cyclopentylfentanyl (10 mg/kg), (±)-cis-3-methylbutyrylfentanyl (0.1, 1, 10 mg/kg), ortho-methylacetylfentanyl (10 mg/kg), para-chloroisobutyrylfentanyl (100 mg/kg), ocfentanil (1, 10 mg/kg), and ortho-fluoroacrylfentanyl (0.1, 1, 10 mg/kg) elicited significant (p ≤ 0.05) dose-dependent increases in locomotion. However, 2,2,3,3-tetramethylcyclopropylfentanyl did not have any effects on locomotion, even when tested up to 100 mg/kg, and 4′-methylacetylfentanyl (10, 100 mg/kg) significantly decreased locomotion. The rank order of efficacy for stimulating locomotion (maximum effect as a % of fentanyl's maximum effect) for fentanyl-related substances relative to MOR agonist standards was cyclopropylfentanyl (108.84 ± 20.21) > fentanyl (100 ± 15.3) > ocfentanil (79.27 ± 16.92) > morphine (75.9 ± 14.5) > (±)-cis-3-methylbutyrylfentanyl (68.04 ± 10.08) > ortho-fluoroacrylfentanyl (63.56 ± 19.88) > cyclopentylfentanyl (56.46 ± 8.54) > para-chloroisobutyrylfentanyl (22.44 ± 8.51) > buprenorphine (11.26 ± 2.30) > ortho-methylacetylfentanyl (9.45 ± 2.92) > 2,2,3,3-tetramethylcyclopropylfentanyl (6.75 ± 1.43) > 4′-methylacetylfentanyl (3.47 ± 0.43). These findings extend in vivo results from previous reports documenting additional fentanyl related-related substances that stimulate locomotion similar to known abused opioids while also identifying some anomalies.

An Endogenous Metabolic Modulator Composition Improves Hydration Status and Decreases Adhesion Properties of Red Blood Cells from Patients with Sickle Cell Disease

Background Sickle cell disease (SCD) is a group of hemoglobinopathies caused by point mutations in the β-globin gene leading to abnormal red blood cells (RBCs). There are various forms of SCD; sickle cell anemia (SCA) is characterized by homozygosity for hemoglobin (Hb) S (HbSS) whereas HbSC disease results from compound heterozygosity for HbS and HbC. HbC does not lead to RBC sickling, but induces loss of intracellular K+ and H2O causing RBC dehydration. Similarly, in HbSS RBCs, through dehydration, an increase of the intracellular concentration of poorly soluble HbS enhances polymerization upon deoxygenation, causing RBCs to become rigid and to sickle. Until recently, the only approved therapy for SCD was hydroxyurea (HU). HU increases fetal Hb (HbF) levels and improves RBC hydration and deformability. For decades, amino acids have been explored as potential treatment to restore the disrupted pathophysiological pathways in SCD. RCitNacQCarLKHVS, an Endogenous Metabolic Modulator (EMM) composition of ten different amino acids and derivatives, was designed to target and support RBC metabolism. In this study, we investigated the effect of ex vivo treatment of this EMM on RBC properties and function in SCA and HbSC. Methods Blood of 12 different adult SCD patients (9 HbSS [of whom 6 were on chronic RBC exchange (RBCX)], 3 HbSC) and 2 healthy controls was collected. Whole blood was incubated at standardized hematocrit (20%, patient plasma) in the presence or absence of different RCitNacQCarLKHVS concentrations for 4h at 37°C. Routine hematological parameters were measured using the Cell-Dyn Sapphire (Abbott Diagnostics). Percentage of total dense RBCs (%DRBCs) was analyzed from 10/12 patients (4 HbSS on RBCX, 3 HbSS not on RBCX, 3 HbSC) using the Advia 120/2120 hematology analyzer (Siemens Healthcare Diagnostics). Point of sickling (PoS), RBC deformability (EImax), and hydration status (Ohyper) were assessed by oxygen gradient ektacytometry and osmotic gradient ektacytometry, respectively, in 12/12 patients using the Lorrca (RR Mechatronics). Blood viscosity was measured in 6/12 patients (3 HbSS not on RBCX and 3 HbSC) at 37°C using a cone-plate viscometer (Brookfield). The hematocrit-to-viscosity ratio (HVR) was calculated as follows: hematocrit/blood viscosity at 300 s-1. RBC adhesion to laminin (BioLamina) was assessed in 4/12 patients (3 HbSS not on RBCX, 1 HbSC) using a microfluidic device (IBIDI μ-Slide VI 0.4). Statistical analysis was performed in Graphpad Prism v9.3.0 using the Friedman test. Results Ex vivo treatment of blood of SCD patients with RCitNacQCarLKHVS showed a significant dose-dependent increase in Ohyper as measured by osmotic gradient ektacytometry (Figure 1), indicating an improvement of RBC hydration status of SCD RBCs. This was accompanied by a significant increase in mean corpuscular volume (MCV) and reduction in mean corpuscular Hb concentration (MCHC). RBCs of healthy controls showed a similar trend in Ohyper, MCV and MCHC. In addition, %DRBCs showed a significant dose-dependent decrease in both HbSS and HbSC RBCs as a result of RCitNacQCarLKHVS treatment. Oxygen gradient ektacytometry showed a significant increase in maximum deformability (EImax), but no significant change in point of sickling. The HVR did not significantly change during dose-response experiments. Median RBC adhesion to laminin decreased as a result of ex vivo treatment with RCitNacQCarLKHVS (Figure 2). Conclusion Ex vivo treatment with RCitNacQCarLKHVS demonstrated a significant dose-dependent increase in RBC hydration (Ohyper, MCV, MCHC and %DRBCs), deformability (EImax) and decreased adhesion properties in SCD RBCs. Dehydrated and dense RBCs with reduced deformability and increased adhesion are more prone to hemolysis, and associated with vaso-occlusive complications such as priapism, skin ulcers, and nephropathy. Results from this study therefore suggest benefit of EMM in modulating clinically-relevant rheological properties such as hydration state and adhesion of SCD RBCs in both SCA and HbSC. The fact that HVR did not change may represent an additional therapeutic benefit, especially for HbSC patients who have, in general, higher blood viscosity compared to HbSS patients. Moreover, it would be worthwhile to investigate if the concomitant use of select amino acid combinations with HU or other anti-sickling agents has an additional beneficial effect in SCD. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Effects of Dolutegravir (DTG) on Survival, Pupariation and Emergence in Drosophila melanogaster: The Rescue Role of Brassica oleracea

The study aimed at determining the protective role of Brassica oleracea on dolutegravir-induced changes in Pupariation and Emergence of Drosophila melanogaster. D. melanogaster aged 3-5 days old were exposed to different concentrations (0.5 to 4 mg/ 5 g diet) of dolutegravir and B. oleracea extract (7.5–1000 mg/5 g diet) for 7 days to determine the lethal concentration (LC50). D. melanogaster were then exposed to the extract (50, 100, 200, and 400 mg/5 g diet) and controls (diet alone and vitamin C) to assess their effects on pupariation and emergence. A 14-day assay was also performed to evaluate the effect of the extract and toxicant (dolutegravir) on fly survival. The result showed a dose-dependent significant decrease (P &lt; 0.05) and a dose-dependent significant increase (P &lt; 0.05) in survival for D. melanogaster exposed to dolutegravir and the extract respectively, when compared to the control group. Results showed a delay in pupariation and decrease in mean pupariation in flies exposed to dolutegravir alone. An improvement in the same parameters was observed in D. melanogaster pre-treated with the extract before exposure to dolutegravir. D. melanogaster pre-treated with 200 and 400 mg extract per 5 g diet showed emergence that was comparable to those in the control groups. A significant decrease (P &lt; 0.05) was observed in the groups exposed to 50 and 100 mg extract per 5 g diet, suggesting no protection at these doses. This study concludes that B. oleracea leaf extract, at certain concentrations, is able to protect against dolutegravir-induced changes in pupariation and emergence in D. melanogaster.