Rodent Model Preclinical Assessment of PEGylated Block Copolymer Targeting Cognition and Oxidative Stress Insults of Alzheimer's Disease.

Misfolded peptide amyloid beta (Aß42), neurofibrillary tangles of hyper-phosphorylated tau, oxidative damage to the brain, and neuroinflammation are distinguished determinants of Alzheimer's disease (AD) responsible for disease progression. This multifaceted neurodegenerative disease is challenging to cure under a single treatment regime until the key disease determinants are traced for their sequential occurrence in disease progression. In an early report, a novel side-chain tripeptide containing PEGylated block copolymer has been tested thoroughly in vitro and in silico for the early inhibition of Aß42 aggregation as well as degradation of preformed Aß42 fibril deposits. The present study demonstrates a preclinical assessment of the PEGylated block copolymer in colchicine-induced AD-mimicking rodent model. The colchicine-induced Wistar rats receiving an intranasal delivery of the block copolymer at a daily dosage of 100 µg/kg and 200 µg/kg body weights, respectively, for 14 days manifested a notable attenuation of behavioral deficit pattern, oxidative stress, and neurotransmitters' deficiency as compared to the untreated ones. The current study also reports the ameliorative property of the PEGylated compound for progressive neuroinflammation and decreased mitochondrial bioenergetics in astrocytoma cell line, viz., U87. A closer look into the drug mechanism of action of a compact 3D PEGylated block copolymer confirmed its disintegrative interaction with Aß42 fibril via in silico simulation. The results obtained from this study signify the potential of the novel PEGylated block copolymer to ameliorate the cognitive decline and progressive oxidative insults in AD and may envision a successful clinical phase trial. The amelioration of disease condition of colchicine-induced AD rat. Initially the rat has given colchicine via stereotaxic surgery which led to a mimicking condition of AD including neuronal death in hippocampal CA1 region. After recovery from the surgery, the rat was treated with the PEGylated block copolymer through intranasal delivery, and this has led to the decrease in neuronal death in hippocampal CA1 region. The mechanism of drug action has shown by the separation of monomer chains of Aß42.

Quercetin: a silent retarder of fatty acid oxidation in breast cancer metastasis through steering of mitochondrial CPT1.

BACKGROUND: Recent evidence confirmed that the maximum energy in metastatic breast cancer progression is supplied by fatty acid oxidation (FAO) governed by a rate-limiting enzyme, carnitine palmitoyltransferase 1 (CPT1). Therefore, the active limitation of FAO could be an emerging aspect to inhibit breast cancer progression. Herein, for the first time, we have introduced quercetin (QT) from a non-dietary source (Mikania micrantha Kunth) to limit the FAO in triple-negative breast cancer cells (TNBC) through an active targeting of CPT1. METHODS: Molecular quantification of QT was confirmed through high-performance thin-layer chromatography (HPTLC). Computational docking analyses predicted the binding affinity of QT to CPT1. Cell-based seahorse energy efflux investigated the mitochondrial respiration rate, glycolytic function and ATP production rate. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) investigated the FAO-associated gene expression. Matrigel cell invasion and fluorescence-activated cell sorting analyses investigated anti-metastatic and apoptotic cell death induction activities, respectively. In vivo antitumor activities were checked using the female breast cancer mice (BALB/c) model. RESULTS: QT resulted in a significant reduction in the intracellular mitochondrial respiration and glycolytic function, limiting extensive ATP production. In turn, QT elevated the reactive oxygen species (ROS) and depleted antioxidant levels to induce anti-metastatic and cell apoptosis activities. qRT-PCR resulted in active healing of altered FAO-associated gene expression which was well predicted through the successful in silico molecular binding potentiality of QT to CPT1. Subsequently, QT has shown excellent in vivo antitumor activities through the altered lipid profile and oxidative stress-healing capabilities. CONCLUSIONS: All the obtained data significantly grounded the fact that QT could be a promising metabolism-targeted breast cancer therapeutic.

Herbometallic nano-drug inducing metastatic growth inhibition in breast cancer through intracellular energy depletion.

Cancer cells need extensive energy supply for their uncontrolled cell division and metastasis which is exclusively dependent on neighboring cells, especially adipocytes. Herein, we have introduced a novel herbometallic nano-drug, Heerak Bhasma nanoparticle (HBNP) from natural resources showing high potential in the reduction of energy supply thereby promoting cell death in breast cancer cells. Inductively coupled plasma optical emission spectra (ICP-OES), atomic absorption spectra (AAS), Raman spectra, X-ray diffraction analyses confirmed the physicochemical properties of HBNP. The differential light scattering (DLS) and field emission scanning electron microscope (FESEM) analyzed the cell-permeable size of HBNP, whereas, cell viability assay confirmed the non-toxic effect. Seahorse energy efflux assay, apoptotic cell quantification, ROS, mitochondrial membrane potential, in vivo oxidative stress etc. were measured using standard protocol. The notable changes in cancer energy metabolism investigated by cellular Mito and Glyco-stress analyses confirmed the HBNP induced intracellular energy depletion. Also, a significant reduction in mitochondrial membrane potential and subsequently, extensive reactive oxygen species (ROS) generations were observed in presence of HBNP followed by the induction of cell apoptosis. The cell invasion and wound healing assay followed by reduced expression both protein (MMP 2, MMP 9) and cytokine (IL6, IL10) had signified the effectiveness of HBNP against cancer metastasis. In addition, HBNP also showed an excellent antitumor activity in vivo followed by developing healing characteristics due to oxidative stress. All these findings strongly suggest that HBNP has the potential to be the new cancer therapeutic. A schematic phenomenon represents the overall HBNP mediated anticancer activity via limitation of both fatty acid uptake and energy metabolism.

A rapid HPTLC method to estimate piperine in Ayurvedic formulations.

BACKGROUND: Trikatu, Sitopaladi, Hingavastaka, Avipattikara, Sringyadi and Talisadya are very popular Ayurvedic (churna) medicines practiced in India; however, unfortunately, they possess several quality control issues. OBJECTIVE: The aim of this study was to find out a simple, accurate and sensitive HPTLC method for the detection and quantification of marker molecule, piperine (alkaloid) on these Ayurvedic formulations for standardization. MATERIALS AND METHODS: Methanolic extraction (reflux) was performed from the above six churnas as well as three single ingredients Piper longum (pipul), Piper nigrum (marich) and Piper chaba (chai). HPTLC was done using piperine as a standard. The mobile phase was a mixture of toluene-ethyl acetate (7:3, v/v) and detection at 342λ. RESULTS: The Rf was detected at 0.39. Piperine was quantified in all samples. P. nigrum showed higher piperine than P. longum and P. chaba. The maximum piperine was noted in Hingavastaka churna and followed by Sringyadi churna, Sitopaladi churna, Talisadya churna, Trikatu churna and Avipattikara churna. CONCLUSION: This method can be successfully employed for standardization and quantitative analysis of piperine in Ayurvedic formulations (churnas) and also be helpful to clinicians and pharmacists to draw significant role of piperine present in all these samples.

Polypharmacy and comorbidity status in the treatment of type 2 diabetic patients attending a tertiary care hospital: An observational and questionnaire-based study.

PURPOSE/AIM: Diabetes mellitus is associated with several comorbid conditions. Thus, often, diabetic patients are prescribed multiple drugs. Although multiple drugs help to combat various diseases, they also increase the propensity of drug interactions and adverse drug reactions. The present study thus tried to evaluate the comorbid conditions and concurrent medications associated with type 2 diabetic patients. It also aimed to address patient compliance for the medications provided to them. MATERIALS AND METHODS: This was a cross-sectional observational study conducted for 2 months - January-February 2017. Data were collected from prescriptions of the patients and also by interviewing the willing patients, attending the Diabetic Clinic of R. G. Kar Medical College, Kolkata, India. RESULTS: During the study period, 150 patients were interviewed and their prescriptions were studied. Out of 150 patients, 69 (46%) were males and 81 (54%) were females. The mean age of the study population was 51.5 (±0.78) years. The present study evaluated that 83.3% (125) of the study population suffered from at least one comorbid conditions, the most common being hyperlipidemia (70.7%) and hypertension (47.3%). The average number of drugs prescribed is 4.72 (±0.11) per prescription. Metformin was prescribed to 96% of the patients. The concurrent medications recommended included hypolipidemics (72%), antihypertensives (68%), drugs for peptic ulcer (34.7%), and antiplatelets (10.7%). CONCLUSION: The present study thus concluded that diabetic patients suffer from a number of comorbid conditions, most commonly, cardiovascular problems. The comorbidity increased with the age. The level of polypharmacy was also high, thereby increasing the pill burden for the patients.

Effect of Coelogyne cristata Lindley in alleviation of chronic fatigue syndrome in aged Wistar rats.

BACKGROUND: Swarna Jibanti scientifically known as Coelogyne cristata Lindley (Orchidaceae), an orchid mentioned in Ayurvedic medicine is used to promote healthy life span. OBJECTIVE(S): The present work was planned to study the efficacy of hydro-alcoholic extract of pseudobulbs of C. cristata (CCE) to assess its role on chronic fatigue syndrome (CFS) induced behavioural and biochemical changes in aged Wistar rats compared to Panax ginseng (PG), a prototype anti-stress agent. MATERIALS AND METHODS: CFS was induced by forced swimming for consecutive 21 days for fixed duration (15 min sessions). The criteria of CFS due to fatigue were counted using locomotor activity, depression and anxiety through automated photactometer, immobility time and plus maze activity respectively. Acute toxicity study of CCE (upto 2 g/kg, Limit test) was also performed. For CFS, animals were divided into five groups, naive control, control, CCE treated (25 mg/kg b.w., 250 mg/kg b.w.) and standard PG treated (100 mg/kg b.w.) groups. All drugs were given orally for consecutive 21 days along with CFS. After assessing behavioural parameters, all animals were sacrificed at day 21 and in vivo antioxidant potential of CCE was determined by lipid peroxides, nitrite, catalase (CAT) and superoxide dismutase (SOD) in brain tissue. RESULTS: CCE was found to be non-toxic. CCE treated aged rats significantly improved (p < 0.001) the spontaneous locomotor movement with respect to control rats, while, decreased the mobility period or depression score. In CFS, CCE also enhanced the time spent (p < 0.001) in open arms while reducing the time spent in closed arm as compared to CFS control, indicating lowering anxiety score. Moreover, marked diminution in lipid peroxidation, nitrite and SOD level was exhibited after CCE treatment and significantly enhanced catalase level significantly (p < 0.01) with respect to CFS control. PG also showed similar actions. CONCLUSION: The results confirmed the potential therapeutic actions of CCE against experimentally induced CFS in aged rats that might be due to its CNS mediatory antioxidant properties.

Antioxidant and hepatoprotective properties of Indian Sunderban mangrove Bruguiera gymnorrhiza L. leave.

BACKGROUND: Bruguiera gymnorrhiza L. (family Rhizophoraceae) is a true mangrove habitat in Indian Sunderban and traditionally uses for liver disorders. OBJECTIVES: The aim was to evaluate antioxidant and hepatoprotective actions of leave extract of B. gymnorrhiza L. MATERIALS AND METHODS: Hydro-methanolic extract of mangrove leaves (BR) was standardized using spectrophotometric and high-performance thin layer chromatography methods. Radical scavenging activities were assessed in different in vitro methods, like 1,1-diphenyl-2-picrylhydrazyl, 2,2'-azino-bis-3-ethyl benzthiazoline-6-sulphonic acid (+), superoxides, nitric oxides and hydroxyl radicals. Hepatoprotective efficacy of BR (125 mg/kg and 250 mg/kg, p.o) was measured in D-galactosamine (GalN) induced (200 mg/kg, i.p) hepatitis in Wistar rats. Silymarin (25 mg/kg, p.o) was used as known hepatoprotective agent. RESULTS: Polyphenols such as gallic acid, quercetin, and coumarin obtained from BR exhibited powerful antioxidant properties. Moreover, it produced dose-dependent protection against GalN induced hepatitis in rats. It significantly reduced GalN induced elevation of enzymes (alanine transaminase, aspartate aminotransferase, and alkaline phosphatase) in serum and resist oxidative stress marked by lipid peroxides, glutathione, and catalase in hepatic parenchyma. CONCLUSIONS: Polyphenols rich B. gymnorrhiza L. leaves ameliorate hepatic tissue injury through its antioxidant effects.

Antidiabetic and antioxidant activity of Rhizophora mucronata leaves (Indian sundarban mangrove): An in vitro and in vivo study.

BACKGROUND: Rhizophora mucronata is a salt-tolerant true mangrove which is widely distributed in Indian mangrove forest and traditionally used to treat diabetes and other health ailments. AIM: The aim of this study is to elucidate the role of Indian variety of R. mucronata leaves on glucose impairing metabolism during diabetes by in vitro and in vivo methods. MATERIALS AND METHODS: The ethanolic fraction of R. mucronata leaves extract (RHE) was assessed for DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging and in vitro anti-diabetic action through α-amylase and α-glucosidase activity assessment. Oral glucose tolerance test (OGTT) and insulin sensitivity test (IST) were assessed and their counteraction with RHE (100 and 200 mg/kg, p.o) and glibenclamide (10 mg/kg, p.o) in streptozotocin (STZ) (50 mg/kg, intravenous) induced hyperglycemic rats were also monitored for 28 days. The data were analyzed statistically using t-test. RESULTS: RHE dose-dependently inhibited α-amylase and α-glucosidase enzymes and lowered the area under the curve (AUC) for glucose on both OGTT and IST. RHE also significantly (p < 0.01) controlled glycemic index and thereby reducing diabetic complications as assessed by lipid profiles, atherogenic index, and coronary index in STZ rats. CONCLUSION: RHE at doses of 100 and 200 mg/kg/day for 28 days provided a significant decrease in diabetes complications and metabolic impairment.

Detection of arecoline by simple high-performance thin-layer chromatographic method in Indian nontobacco pan masala.

Chewing the habit of blended pan masala containing areca nut with or without tobacco is a common practice in the Indian subcontinent. Arecoline, a pyridine alkaloid presence in areca nut alarmed for oral carcinogenesis and strictly prohibited in the western world. However, in India using blended pan masala is very popular among young and old individuals. In this context, we aimed to detect arecoline in Indian blended nontobacco pan masala sold in Kolkata using a simple densitometric high-performance thin-layer chromatographic (HPTLC) method and for alarming their use in common people. Eleven popularly Indian blended nontobacco pan masala were collected from the territory of Kolkata and isolated arecoline, following solvent extraction method derived for pyridine alkaloid. The quantitative analysis of arecoline was measured using automated software-based HPTLC instruments and validated the method according to International Conference on Harmonization guidelines. Arecoline was detected in all 11 blended nontobacco pan masala samples in a range of minimum 130 to maximum 415 µg/g dry samples. Arecoline is hazardous carcinogenic compound, so the use of Indian blended nontobacco pan masala should be restricted. Further, the method was found suitable for routine quantitative analysis of arecoline in areca nut containing substances.

Acute and sub-chronic oral toxicity study of black tea in rodents.

OBJECTIVES: Systematic oral toxicity study for black tea (Camellia sinensis), the most commonly consumed variety of tea, is lacking. The present study was undertaken to assess the iron load on black tea (Camellia sinensis) and its safety aspects in animals. MATERIALS AND METHODS: The analysis of iron was done in six tea samples as per American Public Health Association method using flame atomic absorption spectrophotometer. Maximum physical iron-loaded tea sample was identified on black tea sample 2 (BTS-2), and this was further studied for acute and 90-day sub-chronic toxicity following Organisation for Economic Co-operation and Development guidelines. RESULTS: Black tea sample 2 did not show any signs of toxicity or mortality at up to 2 g/kg per oral dose in Swiss albino mice. 90-day toxicity studies in Wistar rats did not reveal any evidence of toxicity at up to 250 mg/kg/day (2.5% infusion of BTS-2) oral dose as exhibited by regular observations, body weight, food consumption, hematology, serum chemistry, organ weights, and histopathology. Further, serum iron, total iron binding capacity, unsaturated iron binding capacity, and ferritin were not altered after 90 days of treatment. Masson trichrome staining and Perls' staining did not reveal any abnormalities in hepatic tissue following 90-day treatment of high iron-loaded BTS-2. CONCLUSIONS: This safety study provides evidence that BTSs, in spite of relatively high iron content, show no significant iron-related toxicity on acute or sub-chronic oral administration in animals.

Antiradical and antidiabetic properties of standardized extract of Sunderban mangrove Rhizophora mucronata.

BACKGROUND: Mangroves have the ability to grow where no other vascular plants survive. Rhizophora mucronata is a true mangrove and traditionally used to treat diabetes and its allied complications. OBJECTIVES: In the present study, we standardized the 80% methanolic standardized extract of R. mucronata leaves (RH) and found out its antiradical and antidiabetic activities. MATERIALS AND METHODS: The methanolic extract of R. mucronata leaves (RH) was standardized and quantified for phenolics, flavonoinds, gallic acid, quercetin, and coumarin. The reducing abilities and antiradical activities of RH were performed in vitro methods like, 1,1-diphenyl-2-picrylhydrazyl, nitric oxides, superoxides, hydroxyl, and ABTS (2,2'-azino-bis-3-ethyl benzthiazoline-6-sulphonic acid). Thereafter, RH was evaluated for it antidiabetic potentialities on streptozotocin (STZ)-induced type-2 diabetes. STZ (90 mg/kg, intraperitoneal) was administered to 2 days old pups to induce diabetes. RH was fed at doses of 50 and 100 mg/kg and glibenclamide (positive control) at 5 mg/kg, when the rats were 6 weeks old and continued for 10 weeks. Fasting glucose was monitored before and after the treatment. Further, lipid peroxides and reduced glutathione level were estimated on rat liver. RESULTS: The results obtained from this study revealed RH possesses flavonoinds and also gallic acid, quercetin, and coumarin. Further, it has antiradical activities. It has also reduced blood glucose level in type-2 diabetic rats and reduced the formation of lipid peroxidation in liver. RH enhanced the level of glutathione in liver tissue. CONCLUSION: RH exhibits source of natural antioxidants and great potentialities as an antidiabetic agent by improving the hyperglycemia through its antiradical action.

Effects of Indian herbal formulation Body Revival on human platelet aggregation and myocardial ischemia in rats.

OBJECTIVE: To study the effect of Body Revival (BR), a compound traditional Indian herbal medicine, on human platelet aggregation and isoproterenol (IS)-induced myocardial ischemia (MI) damage in male Wistar rats. METHODS: BR suspension 10, 20 and 30 µg was mixed with platelet-rich plasma and incubated at 37 degrees centigrade for 30 min, respectively. Then, adenosine diphosphate (ADP, 20 mmol/L) or collagen (2 µg) was added in the mixture and the aggregation was observed against platelet-poor plasma mixed with equal volume of suspension of the same test samples. Wistar rats divided into 4 groups were used to investigate BR's effects on IS-induced MI. Levels of serum creatinine kinase (CK), aspartate transaminase (AST) and alanine transaminase (ALT) were estimated by standard commercial biological kits. Serum nitric oxide (NOx) was also measured. The lipid peroxides (LPO) and protein concentrations in heart tissues were measured. RESULTS: BR could inhibit ADP- or collagen-induced human platelet aggregation dose-dependently. Moreover, it could protect MI caused by IS in rats. BR reduced the levels of serum CK, AST, ALT and NOx dose-dependently and also lowered LPO in heart tissues in comparison with the MI control (P<0.01). CONCLUSION: BR can inhibit human platelet aggregation and protect MI caused by IS in rats.

Effects of Indian herbal formulation Body Revival on human platelet aggregation and myocardial ischemia in rats / 中西医结合学报

To study the effect of Body Revival (BR), a compound traditional Indian herbal medicine, on human platelet aggregation and isoproterenol (IS)-induced myocardial ischemia (MI) damage in male Wistar rats.

Initial exploratory observational pharmacology of Valeriana wallichii on stress management: a clinical report.

Valeriana wallichii, an Indian medicinal plant, has been on trial for its role in stress disorders in hospital based clinical set-up. Hamilton's Brief Psychiatric Rating Scale (BPRS) was used and thorough clinical investigations were carried out to screen the subjects. Thirty-three subjects (20 male and 13 female; average age 34.2 years) were medicated with the plant extract in a fixed dose regime (500 mg/capsule, twice daily, p.o. after meal). They were thoroughly investigated clinically and using standard questionnaires based on different psychological rating scale at baseline (day 0), mid-term (day 30) and final (day 60). The observations exhibited that, V wallichii not only significantly (p < 0.001) attenuated stress and anxiety, but also significantly (p < 0.001) improved depression and also enhanced the willingness to adjustment. Nevertheless it did not alter memory, concentration or attention of the volunteers. The results suggest that V wallichii may be useful in the treatment of stress related disorders in human and may be a promising anti-stress agent in near future.

Effect of Sonachandi Chyawanprash and Chyawanprash Plus--two herbal formulations on immunomodulation.

The immunomodulatory effects of Sonachandi Chyawanprash and Chyawanprash Plus--two herbal formulations have been evaluated. Both the drugs increased the macrophage activity and their number indicating enhancement of non-specific immune response and reduction of chances of infection. Besides that both Sonachandi Chyawanprash and Chyawanprash Plus efficiently protected Cyclosporine A induced immunosuppression suggesting the immunoprotective role of the aforesaid herbal formulations.

Anti-inflammatory and anti-platelet aggregation activity of human placental extract.

AIM: To find the anti-inflammatory and anti-platelet aggregatory activity of human placental extract (HPE, Placentrex). METHODS: The HPE was studied for anti-inflammatory effect in Wistar rats on carrageenin, serotonin (5-HT), and prostaglandin E1 (PGE1) induced edema in acute model and cotton pellet induced granuloma on sub-acute model. Anti-platelet aggregation was studied against protection of adinosine diphosphate (ADP)-induced aggregation of human platelet through in vitro study. RESULTS: HPE showed positive results both in acute and sub-acute models of inflammation. Highly significant (P<0.01) results were obtained against 5-HT induced acute inflammation and cotton pellet induced sub-acute inflammation in comparison with standard (diclofenac sodium) and control (normal saline) drugs. The anti-inflammatory property of HPE in animal model was well supported with clinical study of platelet aggregation. There was highly significant (P<0.01) inhibition of platelet aggregation with HPE at different doses against ADP. CONCLUSION: Our data suggest that human placental extract may be useful in suppressing inflammation and platelet aggregation.

Studies on the antiinflammatory activity of Betula alnoides bark.

The antiinflammatory activity of Betula alnoides extract was evaluated in acute and subacute inflammation models. The extract was also evaluated for antiinflammatory activity in sheep RBC induced sensitivity and in membrane stabilization models. Except for the sheep RBC induced sensitivity model, the extract showed significant antiinflammatory activity.