Cell Culture and Molecular Docking Analysis to Determine the Antiviral Activity of Folklore Medicinal Plants Against Chikungunya Virus.

Introduction: Chikungunya Virus (CHIKV), a mosquito-transmitted pathogen, poses a significant global health threat owing to its widespread prevalence and high morbidity. There are no approved vaccines or antivirals for prevention or treatment. Screening of folklore medicinal plants has emerged as a promising approach to finding novel therapeutics to combat pathogens. Hence, this study aimed to evaluate the anti-chikungunya potential of folklore medicinal plants and their phytochemicals.

Methods: Maximum non-toxic concentrations (MNTD) of the extracts to Vero cells were determined by the cytotoxicity assay. A Focus-Forming Unit (FFU) assay was used to assess the antiviral activity of the extracts (at MNTD) against CHIKV in Vero cells under pre-, co-, and post-treatment conditions. GC-MS was used to detect the phytochemicals of the extracts, and Schrodinger (Maestro) software was employed for their molecular docking against the target protein of CHIKV.

Results: Azadirachta indica exhibited anti-CHIKV activity during pre- and post-treatment, decreasing the virus titer from 8.145 to 7.998 and 8.361 to 8.040 mean log10 FFU/ml, respectively. Calendula officinalis and Piper retrofractum exhibited anti-CHIKV activity only during post-treatment (8.361 to 8.135, 8.361 to 8.075). Moreover, molecular docking studies of phytochemicals detected in GCMS analysis of all the extracts revealed that many phytochemicals (especially F3, F5, F6, and A1) could bind to the non-structural protein (nSP2) target of CHIKV and suppress the viral replication.

Conclusion: The screened plants showed the ability to inhibit CHIKV infection and replication and hold potential for further investigation in developing treatments for Chikungunya.

Green Synthesized Silver Nanoparticles as Silver Lining in Antimicrobial Resistance: A Review.

Unprincipled use of antibiotics has led to antimicrobial resistance (AMR) against mostly available compounds, and has now become a major cause of concern for the scientific community. However, in the past decade, green synthesized silver nanoparticles (AgNPs) have received greater attention for the development of newer therapies as antimicrobials by virtue of their unique physico- chemical properties. Unlike traditional antibiotics, AgNPs exert their action by acting on multiple mechanisms, which make them potential candidates against AMR. Green synthesis of AgNPs using various medicinal plants has demonstrated a broader spectrum of action against several microbes in a number of attempts. The present paper provides an insight into the scientific studies that have elucidated the positive role of plant extracts/phytochemicals during the green synthesis of AgNPs and their future perspectives. The studies conducted so far seem promising; still, a few factors like the precise mechanism of action of AgNPs, their synergistic interaction with biomolecules, and industrial scalability, need to be explored further till effective drug development using green synthesized AgNPs in healthcare systems against AMR is established.

Demystifying therapeutic potential of medicinal plants against chikungunya virus.

Viral infections are posing a great threat to humanity for the last few years. Among these, Chikungunya which is a mosquito-borne viral infection has produced enormous epidemics around the world after been rebounded. Although this infection shows a low mortality rate, patients suffer from fever, arthralgia, and maculopapular rashes, which reduce the quality of life for several weeks to years. The currently available treatments only provide symptomatic relief based on analgesics, antipyretics, and anti-inflammatory drugs which are nonspecific without satisfactory results. Medicinal plants are a widely accepted source of new molecules for the treatment of infectious diseases including viral infections. The scientific reports, primarily focusing on the anti-chikungunya activity of plant extracts, natural origin pure compounds, and their synthetic analog published from 2011 to 2021, were selected from PubMed, Google Scholar, and Scopus by using related keywords like anti-chikungunya plants, natural antivirals for Chikungunya. The present review decodes scientific reports on medicinal plants against chikungunya virus (CHIKV) infection and demystifies the potential phytoconstituents which reveals that the screening of flavonoids containing plants and phytochemicals showing efficacy against other arbovirus infections, may prove as a potential lead for drug development against CHIKV. The present article also outlines pathogenesis, clinical aspects, molecular virology, and diagnostic approaches of CHIKV infection.

Skin photo-protection with phytochemicals against photo-oxidative stress, photo-carcinogenesis, signal transduction pathways and extracellular matrix remodeling-An overview.

Excessive exposure of skin to UV radiation triggers the generation of oxidative stress, inflammation, immunosuppression, apoptosis, matrix-metalloproteases production, and DNA mutations leading to the onset of photo ageing and photo-carcinogenesis. At the molecular level, these changes occur via activation of several protein kinases as well as transcription pathways, formation of reactive oxygen species, and release of cytokines, interleukins and prostaglandins together. Current therapies available on the market only provide limited solutions and exhibit several side effects. The present paper provides insight into scientific studies that have elucidated the positive role of phytochemicals in counteracting the UV-induced depletion of antioxidant enzymes, increased lipid peroxidation, inflammation, DNA mutations, increased senescence, dysfunctional apoptosis and immune suppression. The contribution of phytochemicals to the downregulation of expression of oxidative-stress sensitive transcription factors (Nrf2, NF-Kb, AP-1 and p53) and protein kinases (MSK, ERK, JNK, p38 MAPK, p90RSK2 and CaMKs) involved in inflammation, apoptosis, immune suppression, extracellular matrix remodelling, senescence, photo ageing and photo-carcinogenesis, is also discussed. Conclusively, several phytochemicals hold potential for the development of a viable solution against UV irradiation-mediated photo ageing, photo-carcinogenesis and related manifestations.

Amantadine exerts anxiolytic like effect in mice: Evidences for the involvement of nitrergic and GABAergic signaling pathways.

Amantadine is a glutamatergic antagonist that works by inhibiting the NMDA receptor. Besides the inhibition of NMDA receptors amantadine also stabilizes the glutamatergic system and protects the neurons against the NMDA toxicity. Amantadine treatment also reduces the production of NO and metabolism of GABA. Therefore amantadine modulates glutamate, GABA and NO which are known to be implicated in the pathogenesis of anxiety and related behavior. The present study was designed to investigate the anxiolytic like effect of amantadine in mice. Nitrergic and GABAergic signaling influence in the anxiolytic like effect of amantadine was also studied. Amantadine (25, 50 and 75 mg/kg, i.p.) was administered and the anxiety related behavior was determined using light and dark box (LDB) and elevated plus maze (EPM) methods. Further, the effect of various treatments on the whole brain glutamate, nitrite and GABA levels were also determined. The results obtained demonstrated that the amantadine (50 mg/kg, i.p.) exerted anxiolytic like effect in mice and reduced the levels of glutamate, nitrite and GABA in the brain of mice as compared to control. Further, the influence of NO and GABA in the anxiolytic like effect of the amantadine was also determined. The results obtained demonstrated that NO donor counteracted while NO inhibitor potentiated the anxiolytic like effect of amantadine in mice. Also the combined treatment of amantadine (25 mg/kg, i.p.) and diazepam (1 mg/kg, i.p.) did not affect the anxiety related behavior, brain GABA and nitrite level of mice but reduced the levels the brain glutamate levels significantly as compared to amantadine (25 mg/kg, i.p.) and diazepam (1 mg/kg, i.p.) treated mice. Thus, amantadine exerted anxiolytic like effect in mice and the anxiolytic like effect of amantadine was modulated by nitrergic and GABAergic signaling pathway.

Lithium potentiated, pyridoxine abolished and fluoxetine attenuated the anxiolytic effect of diazepam in mice.

In the present study, the anxiolytic effect of diazepam (1 and 2 mg/kg, i.p.) was determined alone and in combination with lithium (50 mg/kg, i.p.), pyridoxine (90 mg/kg, i.p.) and fluoxetine (10 mg/kg, i.p.) using elevated plus maze (EPM) and light/dark box (LDB) tests in experimental mice. The effect of various treatments on the brain GABA levels and glutamic acid decarboxylase (GAD) expression were also determined. The results obtained suggested that the diazepam (2 mg/kg, i.p.) exerted anxiolytic effect and significantly increased the brain GABA levels and GAD expression as compared to control group. Fluoxetine (10 mg/kg, i.p.) exerted anxiogenic effects, but did not affect the brain GABA levels and GAD activity significantly as compared to control. Pretreatments of pyridoxine (90 mg/kg, i.p.) abolished; lithium (50 mg/kg, i.p.) potentiated while fluoxetine (10 mg/kg, i.p.) attenuated the anxiolytic and neurochemical effects of diazepam (1 and 2 mg/kg, i.p.) treatment in mice. Therefore, the combined treatment of lithium and diazepam might be a promising treatment for anxiety.

Nitrergic signaling modulation by ascorbic acid treatment is responsible for anxiolysis in mouse model of anxiety.

The present study was designed to investigate the effect of ascorbic acid (AA) treatment on the anxiety related behavioral and neurochemical alterations. AA (50, 100 and 200 mg/kg, i.p.) was administered to the mice and anxiety related behavior and levels of glutamate and nitrite in the brain of mice were determined. The results obtained revealed that the administration of AA (100 mg/kg, i.p.) significantly reduced the anxiety related behavior and the levels of nitrite in the brain of mice. Nitrergic interactions were further determined by the pretreatment of mice with nitric oxide (NO) modulator and AA treatment followed by behavioral and neurochemical measurements. The results obtained suggested that NO inhibition potentiated the anxiolytic like activity of AA in mice. It was also observed that the glutamate and nitrite level in the brain of mice were significantly reduced by the NO inhibitor pretreatment. Thus, the present study demonstrated the possible nitrergic pathways modulation in the anxiolytic like activity of AA in mice.

NO-sGC-cGMP signaling influence the anxiolytic like effect of lithium in mice in light and dark box and elevated plus maze.

Glutamate is an excitatory neurotransmitter implicated in the pathogenesis of psychiatric disorders. Glutamate results in the activation of an enzyme called glycogen synthase kinase-3 (GSK-3) acting through N-methyl-d-aspartate (NMDA) receptors. Impaired expression of GSK-3 affects behavior and neurochemicals level in the brain responsible for the pathogenesis of mood disorders. It has been reported that lithium acts as an inhibitor of GSK-3 and inhibit the enzyme GSK-3 in an uncompetitive manner. In the present study, anxiolytic like effect of lithium in mice is investigated through light and dark box (LDB) and elevated plus maze (EPM). Lithium (50, 100 and 200 mg/kg, i.p.) was administered to the mice to determine the anxiety related behavior. Results obtained suggests that the administration of lithium (100 mg/kg, i.p.) reversed the anxiety related behavior of mice and decreased the levels of glutamate and nitrite as compared to control. Glutamate acting through the NMDA receptor has been found to regulate the expression of enzyme neuronal nitric oxide synthase (nNOS), which is responsible for the release of nitric oxide (NO), suggesting a possible link between NO and GSK-3 also. Therefore, to determine the possible interaction with NO, sub-effective dose of lithium was administered in combination with NO donor i.e. l-Arginine (50 mg/kg, i.p.), NOS and soluble guanylate cyclase (sGC) inhibitor i.e. methylene blue (1 mg/kg, i.p.) and phosphodiesterase inhibitor i.e. sildenafil (1 mg/kg, i.p.). The results obtained demonstrated that the anxiolytic like effect of lithium was abolished by the pretreatment with NO donor and potentiated by the pretreatment with NOS inhibitor. Therefore, it is suggested that NO signaling pathway influence the anxiolytic like activity of lithium in mice, further suggesting the link between the GSK-3 and NO signaling in the regulation of anxiety related behavior.

Anxiolytic-like effect of pyridoxine in mice by elevated plus maze and light and dark box: Evidence for the involvement of GABAergic and NO-sGC-cGMP pathway.

Present study was carried out to investigate the 'anxiolytic-like' effect of pyridoxine in mice. Pyridoxine (90, 180 and 360 mg/kg) was administered by intraperitoneal (i.p.) route to the experimental mice and anxiety-related behavior was evaluated by light and dark box (LDB) and elevated plus maze (EPM) models. Glutamate, GABA and nitrite levels were also determined in the isolated whole brain of mice. It was observed that pyridoxine (180 mg/kg, i.p.) exerted 'anxiolytic-like' effect in mice in EPM and LDB models. Also, there was a significant increase in the levels of GABA whereas; the levels of glutamate and nitrite were decreased as compared to the control group. Administration of pentamethylene tetrazole (PTZ; 20 mg/kg, i.p.) exerted anxiogenic effects in mice, but the combination of PTZ and pyridoxine (180 mg/kg, i.p.) abolished the 'anxiolytic-like' effect of pyridoxine, thereby, suggesting the possible role of GABA in the 'anxiolytic-like' effect of pyridoxine in mice. Further, the influence of NO-sGC-cGMP pathway was investigated by administering the sub-effective dose of pyridoxine in combination with sub-threshold doses of NO modulators i.e. l­arginine (50 mg/kg, i.p.; NO donor); methylene blue (1 mg/kg, i.p.; NO and soluble guanylate cyclase inhibitor) and sildenafil (1 mg/kg, i.p.; phosphodiesterase inhibitor and cGMP modulator). It was observed that the 'anxiolytic-like' effect of pyridoxine in mice was counteracted by the NO donor and potentiated by the NO inhibitors. Thus, the present study confirmed the involvement of GABAergic and NO-sGC-cGMP pathway in the 'anxiolytic-like' effect of pyridoxine in mice.

Antioxidant activity of various extracts of selected gourd vegetables.

Study was conducted to evaluate the antioxidative activity of methanolic (ME), ethanolic (EE) and butanolic extracts (BE) of selected gourd vegetables. The antioxidant activity was investigated using different assays namely ferric thiocyanate test (FTC), thiobarbituric acid test (TBA), ferric reducing antioxidant power (FRAP) and DPPH free radicals scavenging test. A densitometric HPTLC analysis was performed for the analysis of phenolic acids and flavonoids. Different extracts of the selected gourd vegetables revealed different antioxidant activity. Different extracts of Lagenaria siceraria, Momordica charantia and Luffa cylindrica revealed significantly higher (p < 0.05) concentrations of total phenols, flavonids, tannins and carotenoids content and also the antioxidant activity in comparison to remaining vegetable extracts. Correlation studies indicated that FRAP test best described the antioxidant activity of phenols, flavonoids and carotenoids (r = 0.854, 0.692 and 0.915 respectively). HPTLC profiles revealed the presence of maximum number of phenolic acids and flavonoids in L. siceraria and M. charantia.

Cytotoxic potential of few Indian fruit peels through 3-(4,5-dimethylthiazol-yl)-2,5-diphenyltetrazolium bromide assay on HepG2 cells.

OBJECTIVE: To investigate in vitro anticancer activity of a few Indian fruit peels through 3-(4,5-dimethylthiazol-yl)-2,5-diphenyltetrazolium bromide (MTT) assay against HepG2 cells. MATERIALS AND METHODS: Hydroalcoholic extracts were prepared of five fruit peels, i.e., banana, lemon, guava, orange, and papaya by maceration and thereafter subjected for MTT assay to evaluate anticancer potential on HepG2 cells. Plant extract showed best activity was further fractionated with petroleum ether, chloroform, and ethyl acetate successively and screened again. Phytochemical analysis was then carried out to find out responsible components for the observed activity. RESULTS: Out of the 40 samples from five fruit peel extracts with rich folklore usage, papaya extract showed maximum activity with least inhibitory concentration50 (IC50) value of 18.5 µg/ml. Further analysis after fractionation of the papaya peel extract, aqueous fraction showed the maximum inhibitory activity with least IC50 value of 17.3 µg/ml. Phytochemical analysis of the aqueous fraction of papaya peel extract revealed the presence of flavonoids and glycosides. Total flavonoid content found to be 72.25 mg/g. CONCLUSION: Papaya fruit extract demonstrated the best activity against MTT assay which may be due to the presence of flavonoids.

Development and optimization of boswellic acid-loaded proniosomal gel.

CONTEXT: Boswellic acids (BAs) are isolated from oleo gum of Boswellia serrata and are mainly used as potential anti-inflammatory, hypolipidemic, immunomodulatory, and antitumor agents. Pharmacokinetic investigations of BAs uncover its poor bioavailability through digestive system thus creates a need for improved therapeutic responses which can possibly be achieved by developing formulations through novel delivery system. OBJECTIVE: Present study was conducted to design topical BA-loaded proniosomal gel for the management of inflammatory disorders with enhanced bioavailability. MATERIALS AND METHODS: Nonionic surfactant vesicles were prepared using the coacervation phase separation method. A central composite design was employed to statistically optimize formulation variables using Design-Expert software. Three independent variables were evaluated: amount of surfactant (X1), amount of soya lecithin (X2), and amount of cholesterol (X3). The encapsulation efficiency percentage (Y1) and particle size (Y2) were selected as dependent variables. RESULTS AND DISCUSSION: The optimum formulation (F10) displayed spherical bi-layered vesicles under transmission electron microscopy with optimum particle size of 707.9 nm and high entrapment efficiency as 98.52%. In vitro skin permeation study demonstrated the most sustained release of 84.83 ± 0.153 mg/cm2 in 24 h. Anti-inflammatory activity of the gel showed a significant (p < 0.001) higher percentage inhibition as compared to the marketed gel at the same dose. CONCLUSION: The present study exhibited that BA-loaded proniosomal gel was better in terms of absorption, bioavailability, and release kinetics.

Neuropharmacological activities of Taxus wallichiana bark in Swiss albino mice.

AIMS: The bark of Taxus wallichiana is widely used for preparing a decoction and consumed as a tea by several tribal communities of the Indian subcontinent. The sedative, motor coordination, anxiolytic, and antidepressant effects of the hydroalcoholic extract of T. wallichiana bark and its ethylacetate fraction were evaluated in mice models of behavior analysis. MATERIALS AND METHODS: The effects were evaluated on diazepam-induced sleeping time, elevated plus maze and light and dark box, and on the forced swimming test. General locomotor activity and motor coordination effects were evaluated in the actophotmeter and rota-rod tests respectively. STATISTICAL ANALYSIS: Results are expressed as mean ± standard error of the mean. Statistical analysis was performed using ANOVA, followed by post-hoc Dunnett's test. *P < 0.05, **P < 0.01, ***P < 0.001 were considered as significant. RESULTS: Both the hydroalcoholic extract and ethylacetate fraction showed a marked decrease in latency of sleep onset, prolonged the diazepam-induced sleeping time, decreased spontaneous locomotor activity; whereas ethylacetate fraction produced anxiolytic and antidepressant activity. CONCLUSIONS: Both hydroalcoholic extract and its ethylacetate fraction of the bark of T. wallichiana have bioactive principles, which induce neuropharmacological changes.

A review of traditional use, phytoconstituents and biological activities of Himalayan yew, Taxus wallichiana.

Plants synthesize certain phytoconstituents for their protection, which, because they are not of primary need, are known as secondary metabolites. These secondary metabolites of plants, have often been found to have medicinal uses for human beings. One such gymnosperm having secondary metabolites of medicinal potential for humans is Taxus wallichiana (Himalayan yew). Besides being the source of taxol, this plant has been investigated for its essential oil, diterpenoids, lignans, steroids, sterols and biflavonoids. Traditionally, it is used to treat disorders of the digestive, respiratory, nervous and skeletal systems. Although pharmacologically underexplored, it has been used for antiepileptic, anti-inflammatory, anticancer, antipyretic, analgesic, immunomodulatory and antimicrobial activities. The present review compiles traditional uses, phytochemical constituents (specifically the secondary metabolites) pharmacological activities and the toxicity of T. wallichiana.

Cardioprotective effect of ammonium glycyrrhizinate against doxorubicin-induced cardiomyopathy in experimental animals.

OBJECTIVE: The objective of this study was to evaluate the cardioprotective effect of herbal bioactive compound ammonium glycyrrhizinate against doxorubicin-induced cardiomyopathy, in experimental animals. MATERIALS AND METHODS: Ammonium glycyrrhizinate (50, 100, 200 mg/kg, p.o.) was administered for four weeks in albino rats. Cardiomyopathy was induced with a dose of 2.5 mg/kg i.p. of doxorubicin on 1(th), 7(th), 14(th), 21(th), 28(th) day in the experimental animals. At the end of the experiment, on 29(th) day, serum and heart tissues were collected and hemodynamic, biochemical and histopathological studies were carried out. RESULTS: Administration of doxorubicin in normal rats showed significant (P < 0.001) changes in body weight, feed intake, urine output, hemodynamic parameters like (blood pressure, heart rate, cardiac output) and in lipid profile (cholesterol, triglyceride, high density lipoprotein, low density lipoprotein, very low density lipoprotein) indicating cardiomyopathy symptoms. Animals treated with ammonium glycyrrhizinate significantly (P < 0.05) decreased triglyceride, cholesterol, low density lipoprotein (LDL) and very low density lipoprotein (VLDL) levels. Moreover, high density lipoprotein (HDL) levels increased in rats treated with ammonium glycyrrhizinate as compared with the normal group. CONCLUSION: Ammonium glycyrrhizinate is effective in controlling serum lipid profile and cardiac complications in experimentally induced cardiomyopathy in animals.

Submental intubation in panfacial injuries: our experience.

Panfacial fractures present a unique set of problems to the anaesthesiologist and surgeon. Airway management in panfacial fractures is still a challenge to the anaesthesiologist as all modalities available such as orotracheal intubation, nasotracheal intubation, tracheostomy, etc., have their own advantages and disadvantages. When all the conventional modalities to secure airway seem unsuitable then submental route offers an excellent alternative to manage airway in such patients. Here we describe our experience with submental intubation technique in 10 patients with panfacial injuries over a period of two years.

Anaesthetic management of a case of osteogenesis imperfecta with urinary bladder stone - a case report.

SUMMARY: Sometimes in practice of anaesthesia, anaesthesiologist encounters patients with rare congenital diseases. To anaesthesiologist, these patients are a challenge due to inherent complications associated with the disease. Here, we are reporting a case of osteogenesis imperfecta who was posted for the surgery for vesical calculus. All investigations were done to rule out any cardio-respiratory abnormalities, bleeding disorders, which are commonly associated with these patients. Caudal epidural was chosen as anaesthesia technique of choice as spinal anaesthesia was anticipated to be difficult due to associated kyphoscoliosis. GA was avoided due to anticipated difficult airway, restrictive lung disease and susceptibility to malignant hyperthermia. We emphasize the importance of proper preanaesthetic evaluation, intellectual, mental and logistical preparation which should be done before anaesthetising these types of patients.

Antioxidant potential of Lactuca sativa.

The present study is based on the evaluation of antioxidant potential of a well known plant Lactuca sativa. Methanolic leaf extract was investigated for in vitro inhibition of oxidative damage induced by UV-radiations to the salmonella typhi bacteria and in vivo effect on the production of body enzymes i.e. catalase and superoxide dismutase. The lipid peroxidation masurement was also done in terms of thiobarbituric acid reactive substances (TBARS) in blood and brain of male albino wistar rats. The plant extract has shown significant antioxidant potential both in vitro and in vivo.