Ameliorative effect of Nyctanthes arbor-tristis L. by suppression of pentylenetetrazole-induced kindling in mice: An insight from EEG, neurobehavioral and in-silico studies.

Epilepsy is an abiding condition associated with recurrent seizure attacks along with associated neurological and psychological emanation owing to disparity of excitatory and inhibitory neurotransmission. The current study encompasses the assessment of the Nyctanthes arbor-tristis L. methanolic extract (Na.Cr) in the management of convulsive state and concomitant conditions owing to epilepsy. The latency of seizure incidence was assessed using pentylenetetrazol (PTZ) kindling models along with EEG in Na.Cr pretreated mice, trailed by behavior assessment (anxiety and memory), biochemical assay, histopathological alterations, chemical profiling through GCMS, and molecular docking. The chronic assessment of PTZ-induced kindled mice depicted salvation in a dose-related pattern and outcomes were noticeable with extract at 400 mg/kg. The extract at 400 mg/kg defends the progress of kindling seizures and associated EEG. Co-morbid conditions in mice emanating owing to epileptic outbreaks were validated by behavioral testing and the outcome depicted a noticeable defense related to anxiety (P<0.001) and cognitive deficit (P<0.001) at 400 mg/kg. The isolated brains were evaluated for oxidative stress and the outcome demonstrated a noticeable effect in a dose-dependent pattern. Treatment with Na.Cr. also preserved the brain from PTZ induced neuronal damage as indicated by histopathological analysis. Furthermore, the GCMS outcome predicted 28 compounds abundantly found in the plant. The results congregated in the current experiments deliver valued evidence about the defensive response apportioned by Na.Cr which might be due to decline in oxidative stress, AChE level, and GABAergic modulation. These activities may contribute to fundamental pharmacology and elucidate some mechanisms behind the activities of Nyctanthes arbor-tristis.

Neuropharmacological evaluation of different species of Curcubitaceae seeds extract in experimental animals.

Major depressive disorders such as anxiety and depression is predominantly developed in the modern era due to stressful lifestyle and now become the second leading cause of disability. The purpose of the current investigation to evaluate and compared the neuropharmacological effects of three different varaities of Cucurbitaceae seeds including Cucumis Melo var. flexuosus, Cucumis melo var. reticulatus and Santa claus melons ethanol seed extract in experimental animals at three different doses, i.e. 25, 50 and 100mg/kg in animals after 60 days of oral administration. Afterward, various neuropharphamcological activities such as general behavior, phenobarbitone induced sleeping time and exploratory behavior (Elevated plus-maze and head dip test) and motor coordination by Rotarod test were assessed and compared with the control group. The extracts producing dose depended effects on central nervous system. The general behavior profile revealed significant depression at maximum doses. At maximum dose 100mg/kg of Cucumis reticulatus and Santa Claus, seed extracts significantly increases the number of entries in open arms. On the other hand, the Cucumis flexuosus seed extract significantly increases the frequency of numbers of head dips in mice. However, the lower doses of the extracts showed less significant results. The results suggested that all extracts at maximum doses produces anxiolytic effects without affecting the motor coordination in animals.

Mitochondrial dynamics in macrophages: divide to conquer or unite to survive?

Mitochondria have long been appreciated as the metabolic hub of cells. Emerging evidence also posits these organelles as hubs for innate immune signalling and activation, particularly in macrophages. Macrophages are front-line cellular defenders against endogenous and exogenous threats in mammals. These cells use an array of receptors and downstream signalling molecules to respond to a diverse range of stimuli, with mitochondrial biology implicated in many of these responses. Mitochondria have the capacity to both divide through mitochondrial fission and coalesce through mitochondrial fusion. Mitochondrial dynamics, the balance between fission and fusion, regulate many cellular functions, including innate immune pathways in macrophages. In these cells, mitochondrial fission has primarily been associated with pro-inflammatory responses and metabolic adaptation, so can be considered as a combative strategy utilised by immune cells. In contrast, mitochondrial fusion has a more protective role in limiting cell death under conditions of nutrient starvation. Hence, fusion can be viewed as a cellular survival strategy. Here we broadly review the role of mitochondria in macrophage functions, with a focus on how regulated mitochondrial dynamics control different functional responses in these cells.

Quantifying Regulated Mitochondrial Fission in Macrophages.

Mitochondria have co-evolved with eukaryotic cells for more than a billion years, becoming an important cog in their machinery. They are best known for being tasked with energy generation through the production of adenosine triphosphate, but they also have roles in several other cellular processes, for example, immune and inflammatory responses. Mitochondria have important functions in macrophages, key innate immune cells that detect pathogens and drive inflammation. Mitochondrial activity is influenced by the highly dynamic nature of the mitochondrial network, which alternates between interconnected tubular and fragmented forms. The dynamic balance between this interconnected fused network and fission-mediated mitochondrial fragmentation modulates inflammatory responses such as production of cytokines and mitochondrial reactive oxygen species. Here we describe methods to differentiate mouse bone marrow cells into macrophages and the use of light microscopy, electron microscopy, flow cytometry, and Western blotting to quantify regulated mitochondrial dynamics in these differentiated macrophages.

Lipopolysaccharide promotes Drp1-dependent mitochondrial fission and associated inflammatory responses in macrophages.

Mitochondria have a multitude of functions, including energy generation and cell signaling. Recent evidence suggests that mitochondrial dynamics (i.e. the balance between mitochondrial fission and fusion) also regulate immune functions. Here, we reveal that lipopolysaccharide (LPS) stimulation increases mitochondrial numbers in mouse bone marrow-derived macrophages (BMMs) and human monocyte-derived macrophages. In BMMs, this response requires Toll-like receptor 4 (Tlr4) and the TLR adaptor protein myeloid differentiation primary response 88 (MyD88) but is independent of mitochondrial biogenesis. Consistent with this phenomenon being a consequence of mitochondrial fission, the dynamin-related protein 1 (Drp1) GTPase that promotes mitochondrial fission is enriched on mitochondria in LPS-activated macrophages and is required for the LPS-mediated increase in mitochondrial numbers in both BMMs and mouse embryonic fibroblasts. Pharmacological agents that skew toward mitochondrial fusion also abrogated this response. LPS triggered acute Drp1 phosphorylation at serine 635 (S635), followed by sustained Drp1 dephosphorylation at serine 656 (S656), in BMMs. LPS-induced S656 dephosphorylation was abrogated in MyD88-deficient BMMs, suggesting that this post-translational modification is particularly important for Tlr4-inducible fission. Pharmacological or genetic targeting of Tlr4-inducible fission had selective effects on inflammatory mediator production, with LPS-inducible mitochondrial fission promoting the expression and/or secretion of a subset of inflammatory mediators in BMMs and mouse embryonic fibroblasts. Thus, triggering of Tlr4 results in MyD88-dependent activation of Drp1, leading to inducible mitochondrial fission and subsequent inflammatory responses in macrophages.

Preventive and therapeutic effects of aqueous extract of Spinacia oleracea on Psoriatic patches in albino rats.

Psoriasis has become a topic of global concern because of consistency in its prevalence according to Global Report on Psoriasis 2016, however, till date, no therapy has provided complete cure of this disease and no useful measure was discovered to prevent it or eliminate the risk of its relapse. Hence, the present study was designed to evaluate anti-psoriatic effects of Spinacia oleracea due to its anti-inflammatory, anti-oxidant, anti-proliferative and skin-strengthening contents. Psoriasis was induced by oral potassium iodide solution which was then determined by two methods i.e. percentage reduction in psoriatic patch size and psoriasis area and severity index (PASI) score to measure the decrease in severity. Twenty albino rats were used in each method, grouped as standard, curative, preventive and control with five rats in each group. Therapeutic and preventive doses for S. oleracea in both the methods were calculated according to daily intake guidelines of National Cancer Institute, USA, and German Nutrition Society, DGE guidelines respectively with slight modification. Methotrexate was used as standard drug along with folic acid to avoid toxic effects. This study reveals that S. oleracea has both therapeutic and preventive effects. It may be concluded that S. oleracea can be effectively used as sole therapy for psoriasis.

Antihyperlipidemic and anti-hyperglycemic effects of Cymbopogon jwarancusa in high-fat high-sugar diet model.

Hyperlipidemia is the root cause for development of atherosclerosis, coronary heart disease, diabetes mellitus, obesity, hypertension and cerebral palsy. Cymbopogon jwarancusa is an aromatic grass (Rusha grass, khavi grass) belonging to family Poaceaea. C. jwarancusa essential oil is famous for its use in perfumery, soaps cosmetics, detergents, medicine and pharmaceuticals. The anti-pyretic, anti-fungal, antibacterial, anti-oxidant and cytotoxic activities of C. jwarancusa have been reported in literature. In the present study different doses of C. jwarancusa extract have been investigated for anti-hyperlipidemic and anti-hyperglycemic activities in high-fat high sugar diet model in rats. Hyperlipidemia and hyperglycemia were assessed by measuring body weight, serum lipid profile and fasting blood glucose levels. Administration of ethanol leaves extract of C. jwarancusa exhibited significant dose-dependent reduction in body weight, lipid parameters and blood sugar levels. Hence it may be concluded that C. jwarancusa aids in ameliorating hyperlipidemic, hyperglycemic conditions and has potential to reduce the risk of cardiovascular problems.

Report - Antidiabetic effects of native date fruit Aseel (Phoenix dactylifera L.) in normal and hyperglycemic rats.

Change in dietary pattern, sedentary life style and increasing stresses are contributing factors for high prevalence of diabetes mellitus. Diabetic complications often lead to cardiovascular diseases, hypertension and hyperlipidemia that are the leading cause death and disability all over the world. Apart from pharmacotherapy, use of antihyperglycemic medicinal food is a new aspect in diabetes management and prevention of its complications. Phoenix dactylifera (date palm) has been traditionally used for different diseases because of the presence of bioactive agents like anthocyanin, phenols, sterols, carotenoids, procyanidins and flavonoids however its potential as a medicinal food is still unclear. Native date fruit variety Aseel oral suspension was evaluated using 32 normoglycemic and hyperglycemic Sprague dawley rats with two doses (300 and 600mg/kg) against control, disease control and standard drug (Glibenclamide 2.5mg/kg). Outcomes of normoglycemic studies reveals insignificant glucose lowering effects however hyperglycemic studies (glucose challenge and Alloxan inducted hyperglycemia) demonstrates significant glucose lowering effect of Aseel date especially with 300mg/kg dose. Data obtained during this study reveal significant anti-hyperglycemic effects of Aseel dates in diabetes management however further preclinical and clinical studies are required to verify the same.

Prophylactic and therapeutic effect of Punica granatum in trinitrobenzene sulfonic acid induced inflammation in rats.

Pomegranate (Punica granatum L., Punicaceae) contains varieties of antioxidants and phytochemicals; there are evidences that phytochemicals and antioxidants play a vital role in reducing inflammation. Hence this investigation was planned to assess the outcome of Punica granatum on trinitrobenzene sulfonic acid provoked colitis in rats at 2, 5 and 8ml/kg of the body weight. The effect of P. granatum was assessed in two group i.e. prophylaxis as pre-colitis and therapeutic as post-colitis. After completion of dosing in both the groups, macroscopic and histological examination of colon was carried out along with estimation of serum myeloperoxidase, glutathione, alkaline phosphate, fibrinogen and C-reactive protein. In prophylactic procedure P. granatum revealed significant (P<0.05) changes in biochemical markers of inflammation at 5 and 8ml/kg doses. However in therapeutic procedure significant change was observed only at 8ml/kg. Thus results of the present study suggest that P. granatum have a role in prevention as well as treatment of inflammation.

Phytochemistry, Brine shrimp lethality and mice acute oral toxicity studies on seed extracts of Vernonia anthelmintica.

Despite the widespread use of Vernonia anthelmintica seeds in traditional medicine, the need to establish the safety of the Vernonia anthelmintica is required to ascertain the safe use of this herbal medicine. The aim of the present study is to establish the acute toxicity profile of different extracts of Vernonia anthelmintica. Hexane and ethanol extract of Vernonia Anthelmintica has been studied for its brine shrimp lethality potential. Water decoction (WDVA), Hexane (HEVA) and Ethanol (EEVA) extracts of Vernonia anthelmintica has also been evaluated for their in-vivo acute oral toxicity in mice by Lorke's method. Phytochemistry of all three extracts was also evaluated for the presence of their secondary metabolites. All three extracts showed the presence of flavonoids and terpenoids, while alkaloids, tannins and fixed oils were present in HEVA and EEVA. Furthermore EEVA also showed presence of carbohydrates and HEVA also showed the presence of cardiac glycosides. Ethanol and hexane extracts of Vernonia anthelmintica showed a positive cytotoxicity in brine shrimp lethality test at 24 hours with LC50 104.16 (224.0-48.05)µg/ml and 216.11µg/ml (378.2-128.7) respectively as compared to standard drug etoposide LC50 7.46µg/ml. The oral LD50 for EEVA, HEVA and WDVA in mice by Lorke's method was greater than 5000mg/kg. The result of brine shrimp lethality test clearly exhibited the presence of bioactive compounds with cytotoxic potential; however seems to be safe for oral use since LD50 was higher than 5000mg/kg and thus safety of acute dosing in vivo practices is justified.

Differences in lineage replacement dynamics of G1 and G2 rotavirus strains versus G9 strain over a period of 22 years in Bangladesh.

Group A rotaviruses (RVAs) have been a major cause of severe gastroenteritis in Bangladesh, mainly in children below the age of five. At the icddr,b, RVA strains collection and characterization dates back for more than 20 years. This sample collection was used to study the molecular evolution of the VP7 gene of G1, G2 and G9 RVA strains, which have been circulating in Bangladesh for most of this study period. The evolutionary rates (95% HPD) for G1, G2 and G9 were calculated to be 0.93×10(-3) (0.68-1.18), 1.45×10(-3) (1.12-1.78) and 1.07×10(-3) (0.78-1.39), respectively, which is in line with previous data for the RVA VP7 outer capsid protein, which is under strong negative selective pressure. Bayesian analyses revealed that for the G1 and G2 genotypes, one or multiple lineages co-circulated for one or a few seasons, frequently followed by replacement with genetically different lineages. This can be explained by the existence of a large variety of G1 and G2 RVA lineages and the rapid dissemination of different lineages across the globe. In contrast, circulating G9 lineages were rather closely related to each other across the study period and they were usually derived from variants circulating in the previous season(s). This is consistent with the fact that G9 RVAs have circulated in the human population for less than 20 years, and therefore their genetic diversity is much smaller, not resulting in the replacement of circulating G9 strains by highly divergent G9 lineages from abroad. Such different evolutionary dynamics for different RVA genotypes may alter their response to the selective pressure that might be exerted by the introduction of RVA vaccines and therefore a continued close monitoring is warranted.

Cummulative toxicities on lipid profile and glucose following administration of anti-epileptic, anti-hypertensive, anti-diabetic and anti-arrhythmic drugs.

Reporting of undesirable drug reactions is a problem in all countries, even those with sophisticated drug regulatory bodies. However we can expect a horrible picture in developing countries like Pakistan where drug regulatory control is very poor, hence present study has been exclusively designed to explore the outcome of individual administration of antiepileptic, antihypertensive, antidiabetic, antiarrhythmic drugs and their combinations on lipid profile and glucose. The study was conducted on healthy rabbits of either sex. Biochemical tests were performed at the completion of dosing i.e. on 61st day and again after drug-free interval of 15 days. Present study provides detailed evaluation of adverse effects on lipid profile and glucose, results of the study suggests that animals received amiodarone-glibenclamide-verapamil-oxcarbazepine combination did not revealed any significant changes but animals received amiodarone-glibenclamide-losartan potassium-oxcarbazepine and amiodarone-glibenclamide-captopril-oxcarbazepine combinations revealed significant changes. However more studies on large number of animals and human beings are required to justify the use of multiple drug administration, since trial in man is the only way of establishing drug interactions.

Assessment of acute toxicity and reproductive capability of a herbal combination.

The drug under investigation is a herbal combination of Withania somnifera, Tribulus terrestris, Mucuna Pruriens and Argyreia speciosa which has been used for several years of its bio-stimulating, revitalizing and fertility enhancing effects. Present preclinical study is specifically designed to access the safety and efficacy of the product. The result of acute oral toxicity reveals that product is safe up to the dose of 5000 mg/kg. The effects of study related to reproductive capability of drug on both sex reveals increase in reproduction rate up to two generations i.e. F(0) and F(1).

Pharmacological activity of morpholino compound.

The pharmacological effect of morpholino compound in different biological field is of great importance for researchers and investigators. They are working day and night to synthesize pharmacologically more effective morpholino derivatives with less toxic effects. This kind of pharmacological screening of synthesized compounds can lead to the discovery of biologically useful compounds. The findings of the present study are encouraging, since present compound showed a promising analgesic activities and anti-inflammatory. Chemically synthesized morpholino compound are tested for its analgesic and inflammatory activities in intact albino mice ad rat. Compound was administered orally in different doses using the tail immersion test in mice for analgesic activity. The results of this study demonstrated that compound has inhibitory effect on arachidonic acid metabolisms via cyclooxygenase pathway or we can say the inhibition of prostaglandins biosynthesis. Classical non-steroidal anti-inflammatory drug, which inhibits cyclooxygenase, can shift arachidonic metabolism to lipoxygenase products. These findings reflect a new fact of pharmacological action of this compound like inhibition of arachidonic acid pathway in vivo. It can be concluded that the present study revealed a fair success rate but it is necessary to carry out further investigation to confirm the results.