Correction: Shoaib et al. Neuroprotective Effects of Dried Tubers of Aconitum napellus. Plants 2020, 9, 356.

We are sorry to report that some images in Figure 1 reported in our recently published paper [...].

Neuroprotective Effects of Dried Tubers of Aconitum napellus.

The present study was designed to explore the neuroprotective properties of Aconitum napellus (Ranunculaceae). The plant detoxification was done using either water, or cow or goat milk as per the Ayurvedic shodhana method. The evaluation of the neuroprotective role of A. napellus was performed on diabetic neuropathy induced by streptozotocin in Sprague Dawley (SD) rats. Body mass, blood sugar level, oral glucose tolerance test, hyperalgesia, cold allodynia, motor co-ordination test, and locomotor activity, oxidative biomarkers (TBARS, reduced glutathione, catalase and superoxide dismutase) and sciatic nerve histomorphology were assessed. The in vitro studies were done on human neuroblastoma cell line SHSY-5Y and used an MTT assay to assess the antiproliferative activity of different extracts. Results suggest that the goat milk treated chloroform extract has less percentage of aconitine. After administration of the detoxified chloroform extract to the diabetic animals, there was a significant improvement in the myelination and degenerative changes of the nerve fibers along with behavioral changes (p < 0.05 as compared with diabetic control group). The findings of the in vitro research show an effective neuroprotective role of A. napellus. This suggests that A. napellus should be further investigated for its effect in diabetic pathology.

Evaluation of Noxious Consequence of Bark Extract of Onosma echioides Linn Root: Hematology, Biochemistry, and Histopathological Findings.

Onosma echioides is a perennial herb widely used for the treatment of various ailments such as sciatica, gout, and rheumatism. This study focused on toxicological assessment of bark extract of O. echioides root. In subacute toxicity study, 20 Sprague Dawley rats (140 ± 10 g body weight) were randomly grouped into two groups of 10 rats each (5 male and 5 female). Effect of the n-hexane extract of O. echioides (100 mg/kg body weight/day) was studied for a period of 28 days using control and treated groups. Effects of the extract on body weight, food consumption, water intake, serum glucose, and hematology, biochemistry, and histopathology were evaluated. The histopathology was carried out to evaluate the degenerative changes in liver, heart, and kidney. Result of acute toxicity study showed dose-dependent increase in mortality. LD50 was found to be 1,000 mg/kg body weight. The subacute toxicity data showed that the treated group did not show any change in behavior and urinalysis whereas an increase in body weight was observed in the male treated group. A significant but nontoxic effect was observed on food and water consumption. Significant increases in RBC (female treated group; **p < .01), neutrophil (both male and female treated group; **p < .01), MCV (female treated group, **p < .01), MCH (both male and female treated groups; **p < .01), and MCHC (both male and female treated groups; *p < .05) levels were observed; a significant changes were observed in total bilirubin (*p < .05), BUN (**p < .01), potassium (*p < .05), and ALT (**p < .01) levels. The relative organ weights of vital organs at this dose did not show any significant change. In conclusion, the toxicity data showed that the bark extract of O. echioides root does not possess any adverse effect at a fixed dose, which provides a support for its further safety study and biocompatibility application.

Beneficial effects of n-hexane bark extract of Onosma echioides L. on diabetic peripheral neuropathy.

Onosma echioides Linn (Boraginaceae) is the most frequently used curative herb widely used for kidney obstruction, sciatic pain, and gout. The present study was designed to investigate the therapeutic effects of n-hexane bark extract of O. echioides (OE) L. root in vivo against Streptozotocin-induced diabetic neuropathy in SD rats. For in vivo activity, the experiment was categorized into five different groups (n = 5). Group-I was considered as nondiabetic/normal control (NC) treated with 0.5% carboxymethyl cellulose (CMC), Group II as diabetic control, Group-III, IV, and V served as diabetic treated with OE 50, OE 100, and pregabalin at a dose of 50, 100, and 10 mg/kg body weight, orally, respectively. Body weight, blood glucose, oral glucose tolerance test, behavioral studies (motor coordination test, thermal hyperalgesia, cold allodynia, locomotor activity, oxidative biomarkers (thio barbituric acid reactive substances [TBARS], superoxide dismutase [SOD], glutathione [GSH], and catalase), and histopathology of the sciatic nerve were performed. Treatment with OE showed a dose-dependent increase in neuroprotective activity by improving the myelination and decreasing the axonal swelling of nerve fibers. The verdicts of behavioral activities showed a remarkable effect on animals after the treatment of extract and standard drug pregabalin. In conclusion, our findings supported the traditional application of OE and explored its importance in the management of diabetic neuropathy. Additional clinical experiments may provide novel therapeutic drugs for diabetes and its complications.

Cure of human diabetic neuropathy by HPLC validated bark extract of Onosma echioides L. root.

HPLC validated hexane bark extract of Onosma echioides L. root (OE) was evaluated for cure of human diabetic neuropathy in human neuroblastoma cell line. HPLC analysis was performed. Human neuroblastoma cells were grouped into control, normal glucose, high glucose (HG) and HG plus different concentrations of OE extract (10, 25 and 50 µg/mL). MTT, DCFH-DA staining and nuclear condensation assays were performed on neuroblastoma cells to evaluate antiproliferative activity, ROS activity level and apoptotic effect of OE. HPLC analysis revealed the existence of maximum yield of shikonin in n-hexane extract of OE. Exposure with different concentrations of OE effectively decreased ROS level and apoptosis of cells and as a result improved the viability of cells in a dose dependent manner in response to HG-induced oxidative stress. Thus, OE possesses the property to cure human diabetic neuropathy and further can be clinically tested for its use in diabetic neuropathy.

Evaluation of hepatoprotective potential of Erythrina indica leaves against antitubercular drugs induced hepatotoxicity in experimental rats.

BACKGROUND: Erythrina indica Lam. traditionally used in the treatment of laxative, diuretic, worm infestation, liver ailment and joints pain. OBJECTIVE: To evaluate the antihepatotoxic potential of Erythrina indica against isoniazid (INH) and rifampicin (RIF) induced hepatotoxicity in rats. METHODS AND MATERIAL: Liver toxicity was induced by antitubercular drugs (INH+ RIF) at dose level of 50 mg/kg each, p.o for 28 days. 50% methanolic extract of Erythrina indica (100 and 200 mg/kg) were administered orally once daily for 28 days. The hepatoprotective activity was assessed using various biochemical parameters SGOT, SGPT, ALP, bilirubin, total protein, albumin and LDH. Meanwhile, in vivo antioxidant activities as SOD, CAT, GSH and, LPO were measured in liver homogenate also histological examinations were carried out to assess hepatoprotective activity. STATISTICAL ANALYSIS USED: The values were subjected to one way analysis of variance (ANOVA) followed by Tukey multiple compare test. Results were considered statistically significant when P < 0.05. RESULTS: Obtained results demonstrated that the treatment with Erythrina indica (E. indica) significantly prevented drug induced increase in serum levels of hepatic enzymes. Furthermore, Erythrina indica significantly reduced the lipid peroxidation (P < 0.01 tp P < 0.001) in the liver tissue and restored activities of defense antioxidant enzymes GSH (2.15 ± 0.08 to 2.48 ± 0.99; P < 0.05), SOD (2.69 ± 0.752 to 3.712 ± 0.056; P < 0.05 to P < 0.01) and CAT (10.20 ± 0.58 to 12.59 ± 0.42; P < 0.05 to P < 0.001) towards normal. Histopathology of liver tissue showed that Erythrina indica attenuated the hepatocellular necrosis, regeneration and repair of cells toward normal. CONCLUSION: The results of this study strongly indicate the protective effect of Erythrina indica against liver injury which may be attributed to its hepatoprotective activity, and there by scientifically support its traditional use.

Nanoemulsion: for improved oral delivery of repaglinide.

Repaglinide (RPG) is a fast-acting prandial glucose regulator. It acts by stimulating insulin release from pancreatic ß-cells. Recurrent dosing of RPG before each meal is burdensome remedy. Hence the plan of the present study was to evaluate nanoemulsion as a hopeful carrier for RPG for persistent hypoglycemic effect. The drug was incorporated into oil phase of nanoemulsion to give improved biopharmaceutical properties as compared to the lipid-based systems. Pseudo ternary phase diagrams were prepared by aqueous titration method. Formulations were selected at a difference of 5% w/w of oil from the o/w nanoemulsion region of phase diagrams. The optimized nanoemulsion formulation constituted sefsol-218 (5% v/v) as an oil phase, 30% v/v of Tween-80 and transcutol as a surfactant and co-surfactant to restrain nanodroplet size and low viscosity and distilled water (65%). In vitro dissolution studies showed higher drug release (98.22%), finest droplet size (76.23 nm), slightest polydispersity value (0.183), least viscosity (21.45 cps) and immeasurable dilution capability from the nanoemulsion as compared with existing oral tablet formulation. The optimized RPG nanoemulsion formulation showed better hypoglycemic effect in comparison to tablet formulation in experimental diabetic rats. No significant variations were also observed in the optimized formulation when subjected to accelerated stability study at different temperature and relative humidity over a period of 3 months.

Eupalitin induces apoptosis in prostate carcinoma cells through ROS generation and increase of caspase-3 activity.

Prostate cancer is the second most common malignancy in the human reproductive system. Eupalitin is one of the O-methylated flavonol-exhibited enhanced cancer chemopreventive agents. The current study highlights the structural determination of eupalitin and aims to explore the antitumor activity of eupalitin in human prostate cancer cell (PC3) and its underlying mechanism. Eupalitin structure was determined by using FTIR, (1)H NMR, and (13)C NMR. PC3 cells were treated with increasing concentrations of eupalitin, followed by analysis of the cell viability with an MTT assay. The results demonstrated that eupalitin markedly inhibited the proliferation of PC3 cells in a concentration-dependent manner. The results from fluorescent microscopic analysis of nuclear condensation and intracellular ROS generation determined that eupalitin significantly induced ROS level lead to nuclear apoptosis. Cell cycle analysis revealed that eupalitin-induced cell cycle progression as a percentage of cells in G0/G1 phase decreased whereas S phase increased. Caspase-3 immunofluorescence analysis confirms the efficacy of eupalitin-inducing apoptotic pathway and cell death. Thus, our study is helpful in understanding the mechanism underlying these effects in prostate cancer and it may provide novel molecular targets for prostate cancer therapy.

Development and validation of an LC-MS/MS method for simultaneous determination of piperaquine and 97-63, the active metabolite of CDRI 97-78, in rat plasma and its application in interaction study.

Piperaquine-dihydroartemisinin combination is the latest addition to the repertoire of ACTs recommended by the World Health Organization (WHO) for treatment of falciparum malaria. Due to the increasing resistance to artemisinin derivatives, CSIR-CDRI has developed a prospective short acting, trioxane antimalarial derivative, CDRI 97-78. In the present study, a liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method for the simultaneous quantification of piperaquine (PPQ) and 97-63, the active metabolite of CDRI 97-78 found in vivo, was developed and validated in 100 µL rat plasma using halofantrine as internal standard. PPQ and 97-63 were separated using acetonitrile:methanol (50:50, v/v) and ammonium formate buffer (10 mM, pH 4.5) in the ratio of 95:5(v/v) as mobile phase under isocratic conditions at a flow rate of 0.65 mL/min on Waters Atlantis C18 (4.6 × 50 mm, 5.0 µm) column. The extraction recoveries of PPQ and 97-63 ranged from 90.58 to 105.48%, while for the internal standard, it was 94.27%. The method was accurate and precise in the linearity range 3.9-250 ng/mL for both the analytes, with a correlation coefficient (r) of ≥ 0.998. The intra- and inter-day assay precision ranged from 2.91 to 8.45% and; intra- and inter-day assay accuracy was between 92.50 and 110.20% for both the analytes. The method was successfully applied to study the effect of oral co-administration of PPQ on the pharmacokinetics of CDRI 97-78 in Sprague-dawley rats and vice versa. The co-administration of CDRI 97-78 caused significant decrease in AUC0-∞ of PPQ from 31.52 ± 2.68 to 14.84 ± 4.33 h*µg/mL. However, co-administration of PPQ did not have any significant effect on the pharmacokinetics of CDRI 97-78.

Bromelain capped gold nanoparticles as the novel drug delivery carriers to aggrandize effect of the antibiotic levofloxacin.

To develop bromelain capped gold nanoparticles (BRN capped Au-NPs) as the effective drug delivery carriers of the antibiotic levofloxacin (LvN) and evaluate antibacterial potential of its bioconjugated form compared to pure LvN. BRN capped Au-NPs were synthesized by in vitro method and bioconjugated to LvN using 1-ethyl-3-(3-dimethylamino-propyl)-carbodiimide as activator to form Au-BRN-LvN-NPs. These were characterized for mean particle size by dynamic light scattering analysis, zeta potential by Zetasizer nanosystem analysis and transmission electron microscopy (TEM) on carbon coated TEM copper grids by TEM respectively. Drug loading efficiency of LvN was calculated using UV-visible spectroscopy by standard curve of pure LvN. Antibacterial efficacy of Au-BRN-LvN-NPs and pure LvN was determined by evaluating minimum inhibitory concentration (MIC) against Staphylococcus aureus and Eschereschia coli. Two peaks were observed in Au-BRN-LvN-NPs spectrum one at 307 nm and other at 526 nm while one peak in BRN capped Au-NPs at 522 nm during UV spectroscopy suggesting red shift. The drug loading efficiency of LvN was found to be 84.8 ± 2.41 %. The diameter of Au-BRN-LvN-NPs and BRN capped Au-NPs were found to be (58.65 ± 2 nm, 38.11 ± 2 nm), zeta potential (-9.01 mV, -13.8 mV) and surface morphology (~13.2 nm, 11.4 nm) respectively. The MICs against S. aureus and E. coli were found to be (0.128 µg/mL, 1.10 µg/mL) for Au-BRN-LvN-NPs and (0.547 µg/mL, 1.96 µg/mL) for pure LvN. The results suggested that BRN capped Au-NPs can be used as effective drug delivery carriers of the antibiotic LvN. The Au-BRN-LvN-NPs exhibited enhanced antibacterial activity compared to pure LvN alone. (Graphical abstract see Figure 1(Fig. 1)).

New 4'-substituted benzoyl-ß-D glycoside from the fruit pulp of Terminalia belerica with antiplatelet and antioxidant potency.

BACKGROUND: The fruit Terminalia belerica is a rich source of vitamins, acids, and nutraceuticals which have free radical scavenging activity. Thus, the ethanolic extract of fruit and its isolated compound (Tb-01) were intended to estimate antiplatelet and antioxidant activities. METHODS: The ethanolic extract was submitted to Si-gel CC and the compound was isolated. The compound Tb-01 was characterized as benzoyl-ß-D-(4'→10″ geranilanoxy)-pyranosides on the basis of spectral data [ultra violet (UV), infrared (IR), 1H nuclear magnetic resonance (NMR), 13C NMR, and Mass Spectroscopy] and chemical analyses. The ethanolic extract and Tb-01 at different concentrations were in vitro screened for antiplatelet and antioxidant activity. The antiplatelet activity was carried out by using platelet rich plasma prepared by centrifugation of rabbit whole blood (containing 0.9% sodium citrate as anticoagulant) and antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl, reducing power, and nitric oxide anion scavenging activity models. RESULTS: The compound Tb-01 was an amorphous brownish powder, yield 0.64% (w/w), melting point 105-110 °C, Retardation factor/Retention Value (R f value) at 0.42 in methanol:chloroform (20:80) solvent system, UV absorption maxima at 243 nm, and molecular peak [M + H]+ at 394.15 m/z. It was observed that the ethanolic extract and Tb-01 at different concentrations showed significant antiplatelet and antioxidant activity. The antioxidant activity, like scavenging of 1,1-diphenyl-2-picrylhydrazyl radicals, nitric oxide radical, and reductive power were found to be concentration-dependent and increased when increasing amounts of sample were used. CONCLUSION: Mass spectra and 1H NMR confirmed the isolated compound structure which was supported by 13C NMR and IR spectra. Tb-01could be promising for future applications in the treatment of blood clots, pulmonary embolism, and other related diseases.

Effect of methanolic extract of Asparagus racemosus Willd. on lipopolysaccharide induced-oxidative stress in rats.

Lipopolysaccharide (LPS) induced oxidative stress and impairment of normal physiological function generally categorized by increased anxiety and reduced mobility. Therefore, the present study was to find out the effect Methanolic extract of Asparagus racemosus (MEAR ) in lipopolysaccharide (LPS)-induced oxidative stress in rats . LPS-induced oxidative stress in rats was measured by locomotor activity by photoactometer test, anxiety with elevated plus maze test and also studied the oxidative stress markers, nitric oxide and cytokines. The obtained data shows that LPS markedly exhausted (p<0.001) brain- reduced glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) significantly increased (p<0.001) the level of malondialdehyde (MDA), nitric oxide and the activity of cytokines in the brain. MEAR supplementation resulted in normalization of brain GSH and CAT and SOD and decreases in the levels of MDA with reduction of nitric oxide and cytokines in the brain. The action of the extract at dose of 200 mg/kg was almost similar to the standard drug, quercetin (100mg/kg, p.o.). These present study conclude that MEAR administration significantly (P<0.05) reduced LPS- induced oxidative-stress and intensely suggest that Asparagus racemosus Willd. is a functionally newer type of cerebroprotective agent.

Nanomulsion as a carrier for efficient delivery of metformin.

Metformin (MTF) improves hyperglycemia primarily by suppressing glucose production by the liver. The objective of our investigation was to evaluate nanoemulsion as a promising carrier for MTF for sustained hypoglycemic effect. The drug was incorporated into oil phase of nanoemulsion, which finally improved biopharmaceutical properties achieved when compared with lipid based systems. Pseudo ternary phase diagrams were prepared by aqueous titration method. Formulations were selected at a difference of 5% v/v of oil from the o/w nanoemulsion region of phase diagrams, and then thermodynamic stability and dispersibility tests were performed. The composition of optimized formulation was hydrogenated castor oil (5% v/v), 30% v/v of surfactant (tween 80), co-surfactant (transcutol) and distilled water (65% v/v) as an aqueous phase. The preparation showed maximum drug release (98.70%), optimal globule size (92.25 nm), lowest polydispersity value (0.172), lesser viscosity (22.124 cps) and infinite dilution capability. The antidiabetic activity of optimized MTF nanoemulsion formulation evaluated by blood glucose estimation showed significant hypoglycemic effect which was comparable to that observed with conventional marketed formulation in experimental diabetic rats. Optimized formulation was subjected to stability studies at different temperature and relative humidity and was found to be stable. No significant variations were observed in the formulation over a period of 3 months at accelerated storage conditions.

Evaluation of Cucurbita maxima extract against scopolamine-induced amnesia in rats: implication of tumour necrosis factor alpha.

Cucurbita maxima (CM) seed oil is commonly used in Indian folk medicine to treat various ailments. We have investigated the effect of CM seed oil on memory impairment induced by scopolamine in rats. Male adult Wistar rats were administered scopolamine 1 mg/kg body weight, i.p. or 1.25 mg/kg body weight, s.c. to induce memory impairment. The nootropic agent piracetam 100 mg/kg body weight, i.p. and CM seed oil 100 and 200 mg/kg body weight, p.o. were administered daily for five consecutive days. The memory function was evaluated in the Morris water maze (MWM) test, the social recognition test (SRT), the elevated plus maze (EPM) test, and the pole climbing test (PCT). Acetylcholinesterase (AChE) activity and oxidative stress parameters were estimated in the cortex, hippocampus, and cerebellum of the brains after completion of the behavioural studies. The effects of scopolamine on the levels of the tumour necrosis factor alpha (TNF-α) transcript were also investigated. Scopolamine caused memory impairment in all the behavioural paradigms along with a significant increase in the AChE activity and oxidative stress in the brain. Scopolamine also caused a significant increase in the expression of TNF-α in the hippocampus. CM seed oil exhibited antiamnesic activity as indicated by a significant reduction in the latency time in the MWM test and decreased social interaction during trial 2 in the SRT. Further, treatment with CM seed oil significantly decreased the AChE activity and malondialdehyde levels and increased the glutathione level in brain regions. CM seed oil also significantly decreased the expression of TNF-α in the hippocampus. The effect of CM seed oil on behavioural and biochemical parameters was comparable to that observed in rats treated with piracetam. These results indicate that CM seed oil may exert antiamnesic activity which may be attributed to the inhibition of AChE and inflammation as well as its antioxidant activity in the brain.

Evaluation of cardioprotective effect of silk cocoon (Abresham) on isoprenaline-induced myocardial infarction in rats.

OBJECTIVE: The study was conducted to evaluate cardioprotective effect of silk cocoon (Abresham) Bombyx mori (B. mori) on isoprenaline-induced myocardial infarction. This study deals with the cocoons, which is called Abresham in the Unani system of medicine. It is one of the 64 drugs which Avicenna has mentioned in Avicenna's tract on cardiac drugs and used in the treatment of cardiovascular diseases. Abresham is a chief ingredient of the two very famous Unani formulation viz. Khamira Abresham Sada, and Khamira Abresham Hakim Arshad Wala. MATERIALS AND METHODS: The ethanolic extract of B. mori (Abresham) silk cocoons in the dose of 250 mg/kg and 500 mg/kg body weight was administered orally for 28 days before isoprenaline administration to test their cardioprotective effect. Isoprenaline (85 mg/kg) was administered subcutaneously on days 29(th) and 30(th), respectively in order to induce myocardial infarction. RESULTS: The parameters for evaluation of cardioprotective activity were the physical parameters and the biochemical estimations. The physical parameters were gross examination of heart, heart weight/body weight ratio and histopathology examination. In biochemical estimations, the activity of various cardiac enzymes such as aspartate transaminase, alanine transaminase, creatinine kinase, lactate dehydrogenase, and the gold marker troponin-I were determined. The levels altered by isoproterenol were restored significantly by the administration of the both doses of test extract especially at higher dose. CONCLUSION: The result of this study shows that alcoholic extract B. mori has significant cardioprotective activity against isoprenaline-induced myocardial infarction.

Prediction of human absorption of a trioxane antimalarial drug (CDRI 99/411) using an in-house validated in situ single-pass intestinal perfusion model.

OBJECTIVES: The aim of the study was to predict human intestinal permeability and the fraction absorbed of an oral dose of a promising trioxane anti-malarial drug (CDRI 99/411) using the single-pass intestinal perfusion technique (SPIP) in rats. METHODS: Effective permeability coefficients (P eff) in anaesthetized rats were determined for marker compounds and the trioxane derivative 99/411. Drug solution in perfusion buffer was perfused through intestine with a flow rate of 0.2 ml/min and samples were taken from an outlet tubing at different time points up to 120 min. Drug concentrations in samples were determined using RP-HPLC. KEY FINDINGS: The effective permeability coefficient values of marker compounds obtained in rats were compared with published data for human intestinal permeability (P eff (human)) and human fraction absorbed (F(a) (human)) to establish an in-house model. Strong correlations were found between rat and human values for markers (P eff (human) = 1.039 P eff (rat) - 0.1815; R2 = 0.970 and F(a) (human) = 0.15621n (P eff (rat) + 0.7232; R2 = 0.927). Subsequently the human permeability and fraction dose absorbed in human were predicted for 99/411 using the obtained rat permeability value and established correlations. P eff in human predicted from the model was found to be 7.05 x 10(-4) cm/s and F(a) value in human was predicted around 1. CONCLUSIONS: Considering the high correlation of rat Peff values with those of human reported values, it can be concluded that the developed in-house model is reliable and can be used preliminarily, to predict human permeability and fraction dose absorbed of any test compound. From predicted results, 99/411 was found to have high permeability and possibly complete absorption in human.