Significant Dose-dependent Decrease Research Articles

Efficacy of ADIPOR1 and ADIPOR2 peptide-agonist AdipoRon in preventing contracture in a rabbit model of arthrofibrosis.

AdipoRon is an adiponectin receptor 1, 2 (ADIPOR1 and ADIPOR2) agonist with potential antifibrotic effects. Whether AdipoRon can mitigate joint stiffness in a rabbit model of arthrofibrosis is unknown. We examined the efficacy of intravenous (IV) AdipoRon at mitigating contracture in a rabbit model of knee arthrofibrosis. Fifty-six female New Zealand White rabbits were divided into three dosing groups: vehicle (dimethyl sulfoxide, DMSO), 2.5 mg/kg AdipoRon, and 5 mg/kg AdipoRon. AdipoRon, in DMSO, was administered IV preoperatively and for 5 days postoperatively (30 rabbits, Aim 1). AdipoRon was again dosed similarly after Kirschner wire (K-wire) removal at 8 weeks (26 rabbits; Aim 2). The primary outcome of joint passive extension angle (PEA,°) was measured at 8, 10, 12, 16, and 24 weeks following index surgery. At 24 weeks, rabbits were euthanized and limbs were harvested to measure posterior capsular stiffness (N cm/°). In Aim 1, the 5 mg/kg treated rabbits had a significant increase in PEA when compared to controls at 16-week (p < 0.05). In Aim 2, the 5 mg/kg treated rabbits had a significant increase in PEA when compared to controls at 10-week (p < 0.05). In both aims, no significant differences were observed at later time points. Capsular stiffness was no different in any group. We are the first to report the efficacy of IV AdipoRon in a rabbit model of arthrofibrosis. We identified a significant dose-dependent decrease in joint PEA at early time points; however, no differences were observed between groups at later time points. Clinical Significance: The present investigation provided the first assessment of AdipoRon's efficacy in mitigating knee stiffness in the current gold standard rabbit model of arthrofibrosis. Results of this investigation provided further evidence as to the potential role of AdipoRon as a preventative for arthrofibrosis in large mammals.

Effects of individual drug and combination antiretroviral therapy on trophoblast proliferation

BackgroundCombination antiretroviral therapy (cART) has been reported to reduce perinatal transmission of human immunodeficiency virus (HIV) and improve maternal survival outcomes. Recent studies have associated in-utero exposure to cART drugs with adverse outcomes such as pre-eclampsia, preterm delivery, low birth weight and small-for-gestational-age births. However, the exact molecular mechanisms underlying cART-induced adverse pregnancy outcomes remain poorly defined. ObjectivesTo investigate the effects of cART drugs on trophoblast proliferation in the HTR-8/SVneo cell line. Study designHTR-8/SVneo cells were exposed to tenofovir (0.983–9.83 µM), emtricitabine (0.809–8.09 µM) and efavirenz (0.19–1.09 µM), the individual drugs of the first-line single tablet cART regimen termed ‘Atripla’, and zidovudine (1.12–1.12 µM), lamivudine (0.65–6.5 µM), lopinavir (0.32–3.2 µM) and ritonavir (0.69–6.9 µM), the individual drugs of the second-line single tablet cART regimen termed ‘Aluvia’. The cells were treated for 24, 48, 72 and 96 h, and trophoblast proliferation was assessed using a colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltretrazolium bromide assay. ResultsTwo-way analysis of variance showed a significant dose-dependent decrease (p < 0.05) in trophoblast proliferation in response to individual and combined drug components of first- and second-line antiretroviral therapy. ConclusionsFirst- and second-line cART drugs inhibit trophoblast proliferation, and may contribute to placenta-mediated adverse pregnancy outcomes in patients with HIV.

Pioglitazone enhances cisplatin’s impact on triple-negative breast cancer: Role of PPARγ in cell apoptosis

Peroxisome proliferator-activated receptor-gamma (PPARγ) has been recently shown to play a role in many cancers. The breast tissue of triple-negative breast cancer (TNBC) patients were found to have a significantly lower expression of PPARγ than the other subtypes. Furthermore, PPARγ activation was found to exert anti-tumor effects by inhibiting cell proliferation, differentiation, cell growth, cell cycle, and inducing apoptosis. To start with, we performed a bioinformatic analysis of data from OncoDB, which showed a lower expression pattern of PPARγ in different cancer types. In addition, high expression of PPARγ was associated with better breast cancer patient survival. Therefore, we tested the impact of pioglitazone, a PPARγ ligand, on the cytotoxic activity of cisplatin in the TNBC cell line. MDA-MB-231 cells were treated with either cisplatin (40 μM) with or without pioglitazone (30 or 60 μM) for 72 h. The MTT results showed a significant dose-dependent decrease in cell viability as a result of using cisplatin and pioglitazone combination compared with cisplatin alone. In addition, the protein expression of Bcl-2, a known antiapoptotic marker, decreased in the cells treated with cisplatin and pioglitazone combination at doses of 40 and 30 μM, respectively. On the other hand, cleaved- poly-ADP ribose polymerase (PARP) and -caspase-9, which are known as pro-apoptotic markers, were upregulated in the combination group compared with the solo treatments. Taken together, the addition of pioglitazone to cisplatin further reduced the viability of MDA-MB-231 cells and enhanced apoptosis compared with chemotherapy alone.

Evaluation of immunomodulatory (humoral as well as cell-mediated) and cytokines (TNF-α & IL-10) regulating potential of Neolamarckia cadamba fruit extract in Wistar albino rats.

Current work has been designed to investigate the immunomodulatory efficacy with particular reference to humoral, cell-mediated and cytokine-modulating potential of hot aqueous extract of Neolamarckia cadamba (HAENC) fruits in Wistar albino rats. The effect of different concentrations of HAENC fruits over cell-mediated immune response was assessed using six groups (Gp-I as control, Gp-II with 20µg/mL, Gp-III with 50µg/mL, Gp-IV with 100µg/mL, Gp-V with 250µg/mL, and Gp-VI with 500µg/mL) of Wistar albino rats having six animals in each. The amount of tumor necrosis factor-α (TNF-α) and interleukin (IL)-10 was measured by sandwich ELISA with different concentrations of HAENC (50-500µg/mL) in splenocyte culture supernatant and their expression was determined by qRT-PCR Humoral immune response was determined by measuring the serum antibody titer of Wistar albino rats against Salmonella typhimurium 'O' antigen using four groups containing six animals each (Gp-I as control, Gp II, III & IV were respectively fed orally with 125, 250, and 500mg/Kg body weight using HAENC fruits). LC-MS analysis suggested the presence of cadambine, chlorogenic acid, cadambagenic acid, stearic acid, octadecanoic acid ethyl ether, and 7-hydroxy-5,2'-4'-trimethoxyflavonon in the extract based on their m/z ratio. The result suggested significant (p < 0.01) dose-dependent proliferation of Concanavalin A (Con A)-treated splenocytes, depicting cell-mediated immunostimulatory potential of HAENC fruits. A dose-dependent significant decrease (p < 0.01) was found in the amount of TNF-α and IL-10 was found to increase significantly (p < 0.01) as extract concentrations increased. TNF-α and IL-10 expressions were confirmed at the molecular level by qRT-PCR analysis of mRNA transcripts of TNF-α and IL-10 genes. Fold expression of TNF-α and IL-10 gene was 0.197 and 3.58 at 250µg/mL, 0.02 and 20.11 at 500µg/mL concentrations of HAENC respectively in comparison to control. Serum antibody titer was significantly increased (p < 0.01) in animals fed with different doses of HAENC fruits. The present study suggested the anti-inflammatory effect of HAENC fruits which also influences the networking of cytokines, implying that it may play a role in regulating the activity of the host's immune system and can serve as a potent herbal drug with immuno-stimulatory potential.

Assessment of antimalarial activity of crude extract of Chan-Ta-Lee-La and Pra-Sa-Chan-Dang formulations and their plant ingredients for new drug candidates of malaria treatment: In vitro and in vivo experiments.

The emergence and spread of antimalarial drug resistance have become a significant problem worldwide. The search for natural products to develop novel antimalarial drugs is challenging. Therefore, this study aimed to assess the antimalarial and toxicological effects of Chan-Ta-Lee-La (CTLL) and Pra-Sa-Chan-Dang (PSCD) formulations and their plant ingredients. The crude extracts of CTLL and PSCD formulations and their plant ingredients were evaluated for in vitro antimalarial activity using Plasmodium lactate dehydrogenase enzyme and toxicity to Vero and HepG2 cells using the tetrazolium salt method. An extract from the CTLL and PSCD formulations exhibiting the highest selectivity index value was selected for further investigation using Peter's 4-day suppressive test, curative test, prophylactic test, and acute oral toxicity in mice. The phytochemical constituents were characterized using gas chromatography-mass spectrometry (GC-MS). Results showed that ethanolic extracts of CTLL and PSCD formulations possessed high antimalarial activity (half maximal inhibitory concentration = 4.88, and 4.19 g/mL, respectively) with low cytotoxicity. Ethanolic extracts of the CTLL and PSCD formulations demonstrated a significant dose-dependent decrease in parasitemia in mice. The ethanolic CTLL extract showed the greatest suppressive effect after 4 days of suppressive (89.80%) and curative (35.94%) testing at a dose of 600 mg/kg. Moreover, ethanolic PSCD extract showed the highest suppressive effect in the prophylactic test (65.82%) at a dose of 600 mg/kg. There was no acute toxicity in mice treated with ethanolic CTLL and PSCD extracts at 2,000 mg/kg bodyweight. GC-MS analysis revealed that the most abundant compounds in the ethanolic CTLL extract were linderol, isoborneol, eudesmol, linoleic acid, and oleic acid, whereas ethyl 4-methoxycinnamate was the most commonly found compound in the ethanolic PSCD extract, followed by 3-hydroxy-2-(4-hydroxy-3-methoxyphenyl)-4H-chromen-4-one, flamenol, oleic acid amide, linoleic acid, and oleic acid. In conclusions, ethanolic CTLL and PSCD extracts exhibited high antimalarial efficacy in vitro. The ethanolic CTLL extract at a dose of 600 mg/kg exhibited the highest antimalarial activity in the 4-day suppressive and curative tests, whereas the ethanolic PSCD extract at a dose of 600 mg/kg showed the highest antimalarial activity in the prophylactic test.

Antidiarrheal Activities of the Methanol Leaf Extracts of Olinia rochetiana (Oliniaceae) Against Castor Oil-Induced Diarrhea in Mice.

Olinia rochetiana has been used traditionally to cure diarrheal disease. Therefore, this study aimed to investigate the acute toxicity and antidiarrheal effect of O. rochetiana leaf extracts. Cold maceration was used to extract plant leaf powder with 80% methanol. The extract's antidiarrheal action was tested against a castor oil-induced diarrheal model, a charcoal meal test, and enteropooling tests at doses of 100, 200, and 400 mg/kg. Negative controls received the vehicle at 10 mL/kg, while positive controls received loperamide at 3 mg/kg. From the study, no apparent toxicity was observed when a single dose of 2000 mg/kg was administered. In the castor oil-induced model, the extract delayed the onset of diarrhea, reduced stool frequency, and decreased wet feces weight and number in a dose-dependent manner at 200 mg/kg (p < 0.05) and 400 mg/kg (p < 0.01). The percent reduction in moist feces at 100, 200, and 400 mg/kg was 54.2, 23.97, and 18.26%, respectively, indicating a significant dose-dependent decrease. In a charcoal meal test, the extracts at 200 and 400 mg/kg revealed a peristaltic index of 65 and 46%, respectively, with considerable inhibition of charcoal transport at 23 and 39%. The weight and volume of intestinal contents dropped significantly at a dose of 400 mg/kg (p < 0.01), which is 0.43 mg/kg, in the enteropooling test when compared with the tested dose. The computed in vivo antidiarrheal index revealed diarrheal inhibition values of 46.06 and 71.06% at 200 and 400 mg/kg, respectively. In the current investigation, O. rochetiana showed significant antidiarrheal activity with no symptoms of toxicity in mice.

INVESTIGATION OF ANTIANGIOGENIC AND APOPTOTIC EFFECTS OF GALANTAMINE OBTAINED FROM LEUCOJUM AESTIVUM ON MCF-7 BREAST CANCER CELL LINE

OBJECTIVE: The current study aims to investigate the antiangiogenic and apoptotic effects of galantamine in breast cancer cells and to add new information to the literature. MATERIAL AND METHODS: In this study, MCF-7 breast cancer cell line was used and galantamine was obtained from Leucojum aestivum by HPLC method. The effect of galantamine on cell viability was determined by CCK-8 assay at 9 different doses (control, 10, 40, 70, 100, 130, 160, 190, 210 μg/ml) including the control group. Then, to understand the effect of galantamine, other assays (total antioxidant status (TAS), total oxidant status (TOS), poly-ADP ribose polymerase (PARP), and vascular endothelial growth factor (VEGF) levels) were examined in 3 groups; control, LD50 (100 μg/ml) and high dose (210 μg/ml) group. The oxidative stress index (OSI) was then calculated. RESULTS: Galantamine decreased cell viability. The LD50 dose was determined as 100 μg/ml. There was no significant change in VEGF levels. There was a significant dose-dependent decrease in PARP levels (Control: 2.78667±0.155392, LD50: 1.51000±0.107145, High dose: 1.01000±0.054772 ng/L, p=0.000). Compared to the control group, there was a significant dose-related decrease in TAS data (Control: 0.09633±0.002658, LD50: 0.06283±0.002317, High dose: 0.04050±0.001871, p=0.000) and a significant dose-related increase in TOS data (Control: 0.12500±0.010488, LD50: 0.21667±0.015055, High dose: 0.31833±0.021370, p=0.000). A significant dose-related increase in OSI data was found. (Control: 129.964±13.018, LD50: 345.161±26.480, High dose: 788.485±78.575, p=0.000). CONCLUSIONS: It was determined that galantamine had no significant effect on angiogenesis at any dose based on VEGF levels. Galantamine had a negative effect on cell viability and proliferation at certain doses to MCF-7 cells and was found to increase oxidative stress. The decrease in PARP levels indicates the possibility that cells may result in an apoptotic process. These findings may be useful to take a different approach to the use of galantamine in cancer research.

Effect of Combined Methanol Extract of Mucuna Poggei and Justicia carnea Leaves on Hematological Indices and Biochemical Indices of Phenylhydrazine Induced Anaemic Albino Rats

All parts of Mucuna poggei and Justicia carnea have been known to possess valuable medicinal properties. This calls for the need to determine the effect of combined methanol extract of mucuna poggei and justicia carnea leaves on hematological and biochemical indices of phenylhydrazine induced anaemic albino rats. The study was done using 60 male albino rats. The rats were divided into five groups of five (5) rats each. Group 1 (non-anaemic) was administered water and feed only which served as Normal control. Group 2 was administered Phenylhydrazine only which served as negative control. Group 3 through 5 were Phenylhydrazine induced anaemia treated with combined extract (1:1) at 200 mg/kg, 400 mg/kg and 600 mg/kg respectively. The results showed a significant (p&lt;0.05) increase in Hepatic and renal biomarkers (ALT, AST, ALP, Urea, Creatinine) of the anaemic (negative) control. The combined extract significantly restored the alteration through significant (p&lt;0.05) dose dependent decrease in the assayed hepatic and renal function biomarkers. The hematological indices of the treated rats were maintained compared to the anaemic control. The study showed great potential of combined methanol extract of Mucuna poggei and Justicia carnea leaves capable of ameliorating anaemia.

The Designer Drug αPHP Affected Cell Proliferation and Triggered Deathly Mechanisms in Murine Neural Stem/Progenitor Cells.

Increasing reports of neurological and psychiatric outcomes due to psychostimulant synthetic cathinones (SCs) have recently raised public concern. However, the understanding of neurotoxic mechanisms is still lacking, particularly for the under-investigated αPHP, one of the major MDPV derivatives. In particular, its effects on neural stem/progenitor cell cultures (NSPCs) are still unexplored. Therefore, in the current in vitro study, the effects of increasing αPHP concentrations (25-2000 μM), on cell viability/proliferation, morphology/ultrastructure, genotoxicity and cell death pathways, have been evaluated after exposure in murine NSPCs, using a battery of complementary techniques, i.e., MTT and clonogenic assay, flow cytometry, immunocytochemistry, TEM, and patch clamp. We revealed that αPHP was able to induce a dose-dependent significant decrease of the viability, proliferation and clonal capability of the NSPCs, paralleled by the resting membrane potential depolarization and apoptotic/autophagic/necroptotic pathway activation. Moreover, ultrastructural alterations were clearly observed. Overall, our current findings demonstrate that αPHP, damaging NSPCs and the morpho-functional fundamental units of adult neurogenic niches may affect neurogenesis, possibly triggering long-lasting, irreversible CNS damage. The present investigation could pave the way for a broadened understanding of SCs toxicology, needed to establish an appropriate treatment for NPS and the potential consequences for public health.

Modulatory role of dose-dependent quercetin supplemented diet on behavioral and anti-oxidant system in &lt;i&gt;Drosophila melanogaster&lt;/i&gt; model

Background: Quercetin is an abundant bio-flavonoid in foods having several biological activities including anti-oxidant properties. However, the toxicity of quercetin may limit its use. Therefore, screening a graded dose of quercetin may be an important step in its use as a dietary supplement. Hence, the effects of graded doses of quercetin on the survival, locomotive behavior and oxidant-antioxidant system in D. Melanogaster was investigated.Methods: A Survival assay was conducted by feeding 50 fruit flies per vial with 50-400 mg/g quercetin-diet for 14 days. For behavioral and biochemical studies, flies were divided into 4 vials (50 flies each) 0, 100, 200 and 300 mg/g quercetin-diet. Flies were treated with vehicle or quercetin for 7 days. Behavioral assessment of negative geotaxis was determined from the climbing activities of flies. Finally, flies were killed, homogenized and centrifuged for supernatant for the assay of Catalase (CAT), Superoxide dismutase (SOD), Glutathione (GSH), Malondialdehyde (MDA) and Total thiol.Results: Quercetin treated groups showed a dose-dependent significant decrease (p&lt;0.001) in survival rate. Climbing activity was unaltered at 100 mg but significantly (p&lt;0.05) reduced at 200 and 300mg quercetin. Total thiol was not significantly altered (p&gt;0.05) across the groups, MDA was reduced (p&lt;0.05) at 300 mg compared to control, CAT activity was significantly increased (p&lt;0.05) at 100-200 mg compared to control while GSH and SOD at 100-200 mg of quercetin increased.Conclusion: Quercetin showed better antioxidant activity at lower doses and adversely affected the climbing behavior and survival rate at higher doses. Therefore, quercetin use should be dose modulated and used with caution.

Haloperidol's Cytogenetic Effect on T Lymphocytes of Systemic Lupus Erythematosus and Rheumatoid Arthritis Patients: An In Vitro Study.

Investigating haloperidol's cytogenetic behavior in cultured human T lymphocytes of patients with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). Four haloperidol 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, and sister chromatid exchanges (SCEs), proliferation rate index (PRI), and mitotic index (MI) were measured with the optical microscope. Result analysis revealed: (a) a statistically significant (p=0.001) dose-dependent increase of SCEs in SLE patients compared to healthy individuals; (b) a statistically significant (p=0.001) dose-dependent decrease of SCEs in RA patients for haloperidol concentrations 5, 10μg/mL; (c) a statistically significant (p=0.001) dose-dependent increase of SCEs in RA patients for haloperidol concentrations 20, 100μg/mL; and (d) a statistically significant (p=0.001) dose-dependent reduction of PRI and MI in both patient groups compared to healthy individuals. Furthermore, a correlation was observed between (a) SCE and PRI index variations, (b) MI and SCE index variations, and (c) PRI and MI index variations. Haloperidol affects T lymphocytes from SLE and RA patients by modifying DNA replication procedures, DNA damage response, and ferroptosis. Considering the wide use of haloperidol in neuropsychiatric symptoms of SLE and RA patients, further studies with more immune cell subsets are needed to evaluate its effects on human genetic material.

Meldonium Inhibits Cell Motility and Wound-Healing in Trabecular Meshwork Cells and Scleral Fibroblasts: Possible Applications in Glaucoma.

Meldonium (MID) is a synthetic drug designed to decrease the availability of L-carnitine-a main player in mitochondrial energy generation-thus modulating the cell pathways of energy metabolism. Its clinical effects are mostly evident in blood vessels during ischemic events, when the hyperproduction of endogenous carnitine enhances cell metabolic activities, leading to increased oxidative stress and apoptosis. MID has shown vaso-protective effects in model systems of endothelial dysfunction induced by high glucose or by hypertension. By stimulating the endothelial nitric oxide synthetase (eNOS) via PI3 and Akt kinase, it has shown beneficial effects on the microcirculation and blood perfusion. Elevated intraocular pressure (IOP) and endothelial dysfunction are major risk factors for glaucoma development and progression, and IOP remains the main target for its pharmacological treatment. IOP is maintained through the filtration efficiency of the trabecular meshwork (TM), a porous tissue derived from the neuroectoderm. Therefore, given the effects of MID on blood vessels and endothelial cells, we investigated the effects of the topical instillation of MID eye drops on the IOP of normotensive rats and on the cell metabolism and motility of human TM cells in vitro. Results show a significant dose-dependent decrease in the IOP upon topic treatment and a decrease in TM cell motility in the wound-healing assay, correlating with an enhanced expression of vinculin localized in focal adhesion plaques. Motility inhibition was also evident on scleral fibroblasts in vitro. These results may encourage a further exploration of MID eye drops in glaucoma treatment.

Antireproductive changes instigated by efficient drug delivery via papaya seed chloroform extract-based nanoparticles in male rat Bandicota bengalensis

Anti-reproductive potential of papaya seed chloroform extract-based solid lipid nanoparticles (PSCEN) was investigated for the first time in lesser bandicoot rat, Bandicota bengalensis. Mature male rats (n = 30 per group) were fed bait (loose mixture of cracked wheat, powdered sugar, and groundnut oil in the ratio 88:10:2) containing two different concentrations of PSCEN (5% and 10%) in a bi-choice condition for 15 days with one group as vehicle control. The ingestion of active ingredient in 15 days treatment was significantly (P ≤ 0.05) higher by rats treated with 10% PSCEN (39.17–58.70 g/kg body weight) as compared to rats treated with 5% PSCEN (21.30–33.23 g/kg body weight). A dose dependent significant (P ≤ 0.05) decrease was observed in the level of testosterone, FSH, LH and GnRH in plasma of treated rats. A significant (P ≤ 0.05) decrease was also observed in level of total soluble proteins, total lipids, phospholipids and cholesterol in both plasma and testicular tissue, and level of 17β-HSD and 3β-HSD in testicular tissue indicating anti-reproductive effects of PSCEN treatment. There was observed significant (P ≤ 0.05) effect of treatment on histomorphology of testis and cauda epididymis in the form of reduced tubular diameter, germinal epithelial thickness, number of germ cells and dissociation of epithelial cycle in seminiferous tubules, and reduced tubular diameter, increased epithelial thickness, vacuolization, loose contact of principle cells and reduced number of spermatozoa in the cauda epididymal tubules. Maximum antifertility effect was observed with 10% PSCEN treatment, which was not reversed upto 105 days of treatment withdrawal indicating long-term efficacy. The current investigation suggests the use of PSCEN in the management of reproduction of B. bengalensis by exerting influence on testicular and cauda epididymal functions and biochemical parameters.

Safety assessment of Spondias purpurea aqueous leaf extract (anacardiaceae): Acute and sub-chronic toxicity studies in wistar rats

This study aimed to assess the toxicological profile of aqueous leaf extract of S. purpurea (ALES) in Wistar rats. ALES was subjected to acute and sub-chronic toxicity studies which were conducted according to Lorke’s and Organization for Economic Co-operation and Development 408 guidelines respectively. The acute toxicity study was carried out in two phases within 48 hours. Sub-chronic (90 days) toxicity studies were conducted on 4 groups of rats each. The first group received 1mL/kg distilled water, Groups 2, 3 and 4 received ALES 250, 500 and 1000 mg/kg respectively. Preliminary phytochemical screening revealed the presence of saponins, steroids/triterpenes, flavonoids, tannins, alkaloids, and cardiac glycosides. The oral median lethal dose (LD50) of the extract in rats was established to be greater than 5000 mg/kg. Sub-chronic administration of ALES did not produce significant changes in body weights and relative organ weights of treated rats. Hematology results between control and treated groups for a 90-day period of administration of ALES were comparable, however there was a significant (p &lt; 0.05) increase in differential white blood cell at highest dose. Biochemical result, revealed a non-significant mean variations in levels of renal and hepatic parameters between the treated groups and their corresponding controls. However, there was a significant (p &lt; 0.05) dose dependent decrease in level of potassium and glucose for treated groups compared their respective control, also a non-dose dependent decrease in ALP at 500 mg/kg ALES group. Lipid parameters examined also showed a dose dependent decrease in cholesterol, triglyceride, and LDL level, however the decrease was significant for triglyceride at 500 mg/kg and 1000 mg/kg treatment group compared to control. Additionally, in comparison to control a non-significant increase in HDL level was observed for treatment groups. The Histological result showed slight alterations in brain, liver, kidney, lungs and uterus intensified with an increase in the doses of the extracts administered. These findings suggest that Spondias purpurea leaf extract is relatively safe on acute administration. However, long term administration of higher doses could result in mild toxic responses. Therefore, caution should be taken in long term administration of the extract.

Development of Tofacitinib Loaded pH-Responsive Chitosan/Mucin Based Hydrogel Microparticles: In-Vitro Characterization and Toxicological Screening.

Tofacitinib is an antirheumatic drug characterized by a short half-life and poor permeability, which necessitates the development of sustained release formulation with enhanced permeability potential. To achieve this goal, the free radical polymerization technique was employed to develop mucin/chitosan copolymer methacrylic acid (MU-CHI-Co-Poly (MAA))-based hydrogel microparticles. The developed hydrogel microparticles were characterized for EDX, FTIR, DSC, TGA, X-ray diffraction, SEM, drug loading; equilibrium swelling (%), in vitro drug release, sol-gel (%) studies, size and zeta potential, permeation, anti-arthritic activities, and acute oral toxicity studies. FTIR studies revealed the incorporation of the ingredients into the polymeric network, while EDX studies depicted the successful loading of tofacitinib into the network. The thermal analysis confirmed the heat stability of the system. SEM analysis displayed the porous structure of the hydrogels. Gel fraction showed an increasing tendency (74-98%) upon increasing the concentrations of the formulation ingredients. Formulations coated with Eudragit (2% w/w) and sodium lauryl sulfate (1% w/v) showed increased permeability. The formulations equilibrium swelling (%) increased (78-93%) at pH 7.4. Maximum drug loading and release (%) of (55.62-80.52%) and (78.02-90.56%), respectively, were noticed at pH 7.4, where the developed microparticles followed zero-order kinetics with case II transport. Anti-inflammatory studies revealed a significant dose-dependent decrease in paw edema in the rats. Oral toxicity studies confirmed the biocompatibility and non-toxicity of the formulated network. Thus, the developed pH-responsive hydrogel microparticles seem to have the potential to enhance permeability and control the delivery of tofacitinib for the management of rheumatoid arthritis.

Anti-glutamatergic Effects of Three Lignan Compounds: Arctigenin, Matairesinol and Trachelogenin - An ex vivo Study on Rat Brain Slices.

Arctigenin is a bioactive dibenzylbutyrolactone-type lignan exhibiting various pharmacological activities. The neuroprotective effects of arctigenin were demonstrated to be mediated via inhibition of AMPA and KA type glutamate receptors in the somatosensory cortex of the rat brain. The aim of this study was to compare the effects of arctigenin with matairesinol and trachelogenin on synaptic activity in ex vivo rat brain slices. Arctigenin, matairesinol and trachelogenin were isolated from Arctium lappa, Centaurea scabiosa and Cirsium arvense, respectively, and applied on brain slices via perfusion medium at the concentration range of 0.5 - 40 µM. The effects of the lignans were examined in the CA1 hippocampus and the somatosensory cortex by recording electrically evoked field potentials. Arctigenin and trachelogenin caused a significant dose-dependent decrease in the amplitude of hippocampal population spikes (POPS) and the slope of excitatory postsynaptic potentials (EPSPs), whereas matairesinol (1 µM and 10 µM) decreased EPSP slope but had no effect on POPS amplitude. Trachelogenin effect (0.5 µM, 10 µM, 20 µM) was comparable to arctigenin (1 µM, 20 µM, 40 µM) (p > 0.05). In the neocortex, arctigenin (10 µM, 20 µM) and trachelogenin (10 µM) significantly decreased the amplitude of evoked potential early component, while matairesinol (1 µM and 10 µM) had no significant effect (p > 0.05). The results suggest that trachelogenin and arctigenin act via inhibition of AMPA and KA receptors in the brain and trachelogenin has a higher potency than arctigenin. Thus, trachelogenin and arctigenin could serve as lead compounds in the development of neuroprotective drugs.

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.

Curcumin Decreases Viability and Inhibits Proliferation of Imatinib-Sensitive and Imatinib-Resistant Chronic Myeloid Leukemia Cell Lines.

Chronic myeloid leukemia (CML) is a myeloproliferative haematological malignancy characterized by constitutive activation of BCR-ABL1 tyrosine kinase in the majority of patients. BCR-ABL1 expression activates signaling pathways involved in cell proliferation and survival. Current treatment options for CML include tyrosine kinase inhibitors (TKI) with resistance as a major issue. Various treatment options for overcoming resistance are being investigated. Among them, phytochemical curcumin could play an important role. Curcumin has been found to exhibit anti-cancerous effects in various models, including CML, through regulation of multiple molecular signaling pathways contributing to tumorigenesis. We have evaluated curcumin's effects on imatinib-sensitive LAMA84S and K562, as well as imatinib-resistant LAMA84R cell lines. Our results indicate a significant dose-dependent decrease in cell viability and proliferation of imatinib-sensitive and imatinib-resistant cell lines after curcumin treatment. Suppression of key signaling molecules regulating metabolic and proliferative events, such as Akt, P70S6K and NF-kB, was observed. Increased expression of caspase-3 suggests the potential pro-apoptotic effect of curcumin in the imatinib-resistant CML model. Additional in silico molecular docking studies revealed binding modes and affinities of curcumin with different targets and the results are in accordance with in vitro findings. Altogether, these results indicate the potential role of curcumin in the treatment of CML.

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.

SPAD-1, a serine proteinase associated disintegrin from Russell's viper venom disrupts adhesion of MCF7 human breast cancer cells

Serine Proteinase Associated Disintegrin-1 (SPAD-1) is a low molecular mass (26 kDa) positively charged protein purified from Russell's viper venom (RVV) possessing cytotoxic activity on MCF7, human breast cancer cells. Primary sequence analysis of the protein confirms that it is a novel Snake Venom Serine Proteinase (SVSP) and a member of the trypsin family. SPAD-1 contains a conserved triad of Histidine (H), Aspartic acid(D) and Serine(S) residues at its active site for proteinase activity and also an adjacent histidine-glycine-aspartic acid (HGD) disintegrin-like motif. The serine proteinase and disintegrin parts are functionally active and independent. SPAD-1 showed proteolytic digestion of fibrinogen and fibronectin, but laminin digestion was below the detectable limit. Proteolytically inactivated SPAD-1 inhibited collagen and ADP-induced platelet aggregation. This study proposes considering Serine Proteinase Associated Disintegrin (SPAD) as a new group of snake venom proteins. Members of this group contain a serine proteinase catalytic triad and a disintegrin-like motif. SPAD-1 caused visible morphological changes in MCF7 cells, including a reduction of the cell-to-cell attachments, rounding of cell shape and death, in vitro. SPAD-1 also showed a dose-dependent significant decrease in the invasive potency of breast cancer cells. Confocal microscopic analysis revealed the breakage of nuclei of the SPAD-1-treated cells. SPAD-1 also increased cell detachment from the poly L-lysine-coated, laminin-coated and fibronectin-coated culture plate matrices, confirming the disintegrin activity. This study concludes that SPAD-1 may be a good candidate for anti-tumour drug design in the future.