Effects of ciprofloxacin or co-amoxicillin with and without antidepressants on behavioral deficit in rats induced with chronic suppurative otitis media.

Chronic suppurative otitis media (CSOM) is chronic infection of middle ear which is usually managed with antibiotic therapy. This infection may cause the depression and cognitive changes in patients. The aim of present study was to evaluate the effect of antibiotic (ciprofloxacin and co-amoxicillin) with antidepressant or without antidepressants (bromazepam and imipramine) at low doses on rats with induced with CSOM. Pseudomonas aeruginosa was used to induce CSOM by in rat ear (tympanic bulla). The rats were divided into eight groups having six animals in each group. Neuropharmacological activities and gross behavior were observed in open field activity, force swimming cage, maze test, light and dark activity box and traction test. Observations were noted weekly after the administration of ciprofloxacin (15.3mg/kg), co-amoxicillin (15.3mg/kg), imipramine (1.15mg/kg) and bromazepam (0.09mg/kg) intraperitoneally. The altered behavior and depression was observed in control positive but reverted back in groups maintained on antidepressants with antibiotics with significant improved locomotor activity, memory in memory cage, muscular co-ordination and body balance and decreased anxiety. On the other hand, groups treated with only antibiotics showed significant improvement only in force swimming and traction test at day 14. Therefore, the antidepressant effects of the drugs can be employed to attenuate stress and depression in patients with CSOM.

Anti-inflammatory and histopathological studies of leaves extracts of Croton bonplandianus in Albino rats.

Non-steroidal anti-inflammatory drugs (NSAIDs) are frequently used for the treatment and prevention of inflammation with the increase in number of side effects. Traditional plants have been used to treat inflammation owing to lesser adverse responses. Croton bonplandianus being an anti-inflammatory plant is extensively utilized all over the world. The methanolic and aqueous leaves extracts of Croton bonplandianus were exposed to anti-inflammatory activity in the carrageenan induced paw edema against standard diclofenac sodium, followed by the histopathlogical examination. The highest dose of methanolic extract were shown significant anti-inflammatory action having a significant P-value (P<0.05-0.001) compared with the diclofenac sodium (P<0.01-0.001) and aqueous extracts (P<0.5-0.01). The histopathological examination illustrated the vasodialation with reduction in the intensity of edema, neutrophils infiltration and other inflammatory cells. C. bonplandianus being a reactive oxygen species scavenger, responsible to exert an excellent anti-inflammatory activity. The present study confirmed the anti-inflammatory potential of drug extracts and authors recommended its utilization in the treatment of pain, inflammation and relevant diseases in future. However, phytochemical screening is to be required for the complete evaluation of active chemical constituent (s).

GC/MS assisted phytochemical analysis of Ajuga parviflora leaves extract along with anti-hepatotoxic effect against anti-tubercular drug induced liver toxicity in rat.

Owing to its traditional applications, the current study focuses on Ajuga parviflora (A. parviflora) leaves extract for phytochemical and pharmacological analysis. The principle constituents were identified through gas chromatography (GC), and gas chromatography/mass spectroscopy (GC/MS), these includes phthalic acid, squalene, α-tocopherol, vitamin E, phytol, 2-methylenecholestan-3-ol, stigmasterol, cholest-22-ene-21-ol and 3,5-dehydro-6-methoxy. Hepatoprotective effect of A. parviflora was evaluated through isoniazid and rifampicin (INH and RFP) induced hepatotoxicity in rat. Animals in group A were treated with INH and RFP 50 mg/kg. Animals in group B, C, and D were pre-treated with A. parviflora extract at 100, 200 and 300 mg/kg dose prior drug administration. A. parviflora extract at 200 and 300 mg/kg in group C and D significantly reduced aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and bilirubin (p<0.001) as compare to group B (100mg/kg). Total protein (TP) was also significantly (p<0.01) reduced in group C and D at dose of 200 and 300 mg/kg, respectively. The extract pre-treated animals with (A. parviflora, 200, and 300 mg/kg) showed that the epithelium of the central portal vein is intact with replete glucagon. The pre-treatment with A. parviflora protected the liver from INH and RFP induced hepatotoxicity. The results of pre-treated animals with A. parviflora 200, and 300 mg/kg dose prettily revert the severely disturb parameters like, cytolysis, lymphocytic infiltration, and lymphoid aggregate in portal vein and hydropic degeneration. The decrease peroxisome proliferator-receptor activator-δ (PPAR-δ) gene expression by INH, and RFP was significantly up regulated by A. parviflora extract in pre-treated animals at 200 and 300 mg/kg dose. These findings provide baseline pharmacological uses of A. parviflora in liver disorders. Further investigations are required for identification and isolation of biologically active components responsible for pharmacological activity.

GC-MS profiling, FTIR, metal analysis, antibacterial and anticancer potential of Monotheca buxifolia (Falc.) leaves.

Monotheca buxifolia has traditionally been employed in folk medicines to cure of infectious diseases. Current study was aimed to standardize the M. buxifolia leaves extract and evaluate its antibacterial and anticancer activity. Phytochemical analysis was carried through GC, GC/MS, FTIR, and ICP-OES analytical techniques. Antibacterial assay of the crude extract was performed by using tetrazolium micro plates. The extract treated bacteria were observed under (AFM) atomic force microscope and PCR was used for DNA amplification. The anti-proliferative activity of M. buxifolia leaves extract was examined through MTT cytotoxicity assay. The bacterial strains employed in this study were S. epidermidis ATCC (13518), S. aureus ATCC (25923), P. aeruginosa ATCC (10145), and E. coli ATCC (10536). Minimum inhibitory concentration (MIC50) against gram positive bacteria was significantly (p<0.01) achieved at 50 and 75µg/mL. MIC50 against E. coli and P. aeruginosa was also significant at 100µg/mL (p<0.01). M. buxifolia leaves extract damaged the cell walls gram-positive and gram-negative bacteria, while biofilm around gram positive bacteria was significantly damaged. The DNA decantation was also inhibited of S. aureus and S. epidermidis, however, no any impact was observed on E. coli and P. aeruginosa DNA decantation. The cytotoxicity findings suggested that the crude extract of M. buxifolia leaves at 1000µg/mL gives significant inhibition 73.96±2.0%, 83.76±1.2%, 77.66±1.2% and 72.67±1.6% against MDA-MB-231, MCF-7, HeLa and H460 cell lines respectively at (p<0.001). It may be concluded that M. buxifolia leaves extract have significant and promising antibacterial and anti-cancer activities which could be helpful to establish new antimicrobial and anticancer agents.

Phytochemical screening and assessment of analgesic, Anti-inflammatory and hematological properties of the fruit of Berberis baluchistanica.

Traditionally Berberis species have been used as anti-inflammatory, anti-rheumatic, analgesic and anti-anemic drugs. This study was aimed to determine chemical constituents and to assess analgesic, anti-inflammatory and hematological effects of the crude extract of the berries of Berberis baluchistanica to verify these folkloric claims. Phytochemical screening, carried out by using different chemical reagents and techniques like Thin Layer Chromatography (TLC) and Fourier Transform infra-Red (FTIR) indicated presence of flavonoids, saponins, phytosterols and carbohydrates including reducing sugars. Analgesic and anti-inflammatory activities were assessed on mice by using acetic acid induced writhing method and formalin method. Potent anti-inflammatory and analgesic effects were observed during these experiments. The extract also showed anti anemic effect as it increased the levels of hemoglobin and red blood cells significantly. Increase in the platelet count was also noted. The extract of the berries was used at oral doses of 300 and 500 mg/kg during experiments. Anti-inflammatory and analgesic activities were determined by comparing with the standard i.e. aspirin 300 mg/kg. Both doses produced significant anti-inflammatory and analgesic activities at P<0.05. These activities were seemingly attributable to flavonoid and saponin contents of the drug. These results justify the folkloric claims that the drug could be used as good anti-inflammatory, antirehumatic, analgesic and anti-anemic drug. However, further chemical investigations on the drug are suggested for isolation and identification of compounds that could be safer and more effective than the currently available medicines in treating these disorders.

Phytochemical screening and anti-oxidant activity of the two plants Ziziphus oxyphylla Edgew and Cedrela serrata Royle.

Phytochemical studies of medicinal plants are a basic and helping tool for the isolation of active secondary metabolites. The isolation of active compounds is made easy by the help of preliminary phytochemical studies, which shows the presence of a specific class or group of compounds present in these medicinal plants. Ziziphus oxyphylla and Cedrela serrata are medicinal plants with valuable local uses. The present study is for the first Phytochemical investigation of these two medicinal plants which consists of, Quantitative tests showing very good results except Ziziphus oxyphylla plants which does not showed the results for Ester value and Peroxide value. Color reactions are studied for all the crude extracts showing the presence of a number of chemical groups belonging to the class of Alkaloids, Phenol compounds, Phenothiazines, Aromatic compounds, Amino acids, Sulfur compounds etc. Brine shrimp activity was performed which showed a LD50 value of 45.74 and 53.36 in the case of Ziziphus oxyphylla roots and Cedrela serrata bark respectively, which is comparable to the standard drug Cyclophosphamide results of 16.09. Insecticidal activity did not show any promising result indicating the absence of any insect killing potency. Antioxidant activity was very positive for all the extract particularly, the Ziziphus oxyphylla roots, which showed even better results than the standard drug Ascorbic acid used in various dilutions.

Anti-dermatitis, anxiolytic and analgesic effects of Rhazya stricta from Balochistan.

Current study was carried out on Rhazya stricta. Plant material was collected from Jhalmagsi Dist. Balochistan, Pakistan. Methanolic extract of Rhazya stricta was tested for anti-dermatitis, analgesic, anxiolytic effects, insecticidal activity and Brine shrimp Bioassay. Crude extract showed significant anti-dermatitis activity, as the results of intensity score showed mild Excoriation or erosion, moderate Edema or populations and absence of Erythema or hemorrhage, Scratching time was decreased to 1.45 and histological observations of mice treated with crude extract showed mild changes and few inflammatory cells in several microscopic fields. The results of analgesic activity were significant and the percentage inhibition of writhes were 73.54% and 69.38% at 300mg/kg and 500mg/kg respectively. The overall response of crude extract in anxiolytic activities were depressive and crude extract showed sedative effects. In Brine shrimp (Artemsia salina) lethality bioassay crude extract showed dose depended significant activity, and showed positive lethality with LD(50) 3.3004µg/ml. Insecticidal activity was positive against Callosbruchus analis, the percent mortality was 40%.

Phytochemical and pharmacological studies on methanolic seeds' extract of Dolichos biflorus.

The Dolichos biflorus is a well known medicinal plant in folklore for its medicinal properties. In herbal medicine the seeds of it are mainly used as tonic, astringent, diuretic, and are also recommended in asthma, bronchitis, urinary discharges, hiccoughs, ozoena, heart trouble and other diseases of brain. The main purpose of this study is to explore and to provide experimental data on the traditional use of plant Dolichos biflorus. For this purpose we investigated the plant seed extract phytochemically and pharmacologically. Phytochemical analysis was performed on extract and powder form of the drug. Procedure use for evaluation were Identification of chemical constituent by color reaction, Fluorescence analysis of powder drug, pH (in powder and extract forms), loss on drying, Thin layer chromatography, Infrared spectroscopy, acid and saponification values. In pharmacological studies (diuretic, analgesic and anti-inflammatory activities) were tested on the extract of plant seed. The tests were carried out over albino mice taking different concentration of seed extract. Seeds extract of Dolichos biflorus has exhibited mild analgesic activity, the results were (84.6±6.68) at dose 300mg/kg and (92.2±6.81) at dose 500mg/kg which were not much significant as compared to reference drug Aspirin (300mg/kg) having result (36.4±2.27). While seed extract of Dolichos biflorus exhibited remarkable diuretic activity, the values at 300 mg/kg was (1.33±0.13) and at 500 mg/kg were (2.66±0.31) which are highly significant as compared to drug Lasix (20mg /kg) having result (2.38±0.23). Anti-inflammatory effects of crude extract of Dolichos biflorus obtained at 0.06mg/kg and 01mg/kg were (26.6±2.96) and (36±1.67) respectively. While the value for aspirin as standard drug (300mg/kg) were (17.44±1.59).This study provides a platform for further investigation for the isolation of active principles responsible for biological activity.

Toxicity studies on herbal formulation used in diabetes mellitus.

In current study herbal formulation was prepared for Diabetes mellitus (type 2). It consists of the extracts of Salacia reticulate, Cinnamomum zeylanicum,Lagerstroemia speciosa, Camellia sinensis and Gymnema sylvester. Toxicitystudies were carried out on heart, liver, kidney and blood of both male and female rabbits. Drug was administered in a dose of 15mg/kg body weight daily for 90 days. On 91th day, blood was drawn from animals and investigated for changes in biochemical and hematological levels. After that animals were sacrificed and their organs (liver, heart and kidney) were analyzed for histo-pathological changes. In biochemical tests for lipid profile, significant decreased (male-70.64 ± 0.321; female-69.80 ± 0.365) in triglycerides level were observed, no significant change was recorded in Cholesterol HDL ratio, LDL, VLDL level. A significant increase (male-16.00 ± 1.418; female-10.00 ± 0.709) was observed in HDL level. In liver function test significant decrease was observed in Gamma GT (male-10.08 ± 0.862; female-7.00 ± 0.709). Alkaline phosphatase (male-79 ± 0.838; female-51.1 ± 1.810), SGPT (male-54 ± 0.709; female-43.04 ± 2.060), direct bilirubin (male-0.024 ± 0.005; female-0.014 ± 0.002) and total bilirubin (male-0.109 ± 0.003; 0.106 ± 0.049) were observed. Non-significant changes were observed in serum total protein, globulins, albumin and A/G ratio. No significant changes were noticed in urea level and serum electrolytes. In cardiac enzymes significant decrease was observed in LDH (male-443 ± 5.61; female-360 ± 1.848) and SGOT (male-27 ± 0.709; female-28 ± 1.418) level and highly significant rise in CPK (male- 3128 ± 8.478; female-1598 ± 7.483) and CK-MB (male-446 ± 2.308; female-438 ± 2.819). In hematological profile, significant decrease was observed in Hb (male-12.3 ± 0.392; female-12.4 ± 0.1), RBC count (male-6.60 ± 0.167; female-5.74 ± 0.25) and Hematocrit (HCT/PCV) % in both male and female rabbits (male-45.70 ± 0.255; female-43.50 ± 0.448) and significant (p<0.5) increased in WBC count (male-8.40 ± 0.401; female-9.10 ± 0.054). Significant (p<0.5) decrease in blood glucose level and HbA1c (male-3.36 ± 0.113; female-3.16 ± 0.076) was observed. In histopathological studies mild edema was observed in heart and there was no change in histo-architecture of liver and kidneys. It is concluded that formulation does not showed any chronic toxicity in adult dose.

Evaluation of phytoconstituents of three plants Acorus calamus linn. Artemisia absinthium Linn and Bergenia himalaica boriss by FTIR spectroscopic analysis.

Qualitative and quantitative analysis of plant extracts can be achieved by using different spectroscopic techniques. In current research work we deal with the nature of the absorption and spectra of extract of Acorus calamus, Artemisia absinthium and Bergenia himalaica using FTIR spectroscopic technique. The present study was focused on standardization of crude extracts by utilization of infrared light. The spectra of crude extracts (A. calamus, A. absinthium and B. himalaica) displayed very clear diagnostic peaks of functional groups i.e. O-H alcoholic/acid, C-H alkyl & aromatic ring, carbonyl, and C-O-C groups. The spectra of all the three plants did not show any peak at 2220-2260 cm(-1), which is indicative of the absence of nitrogen containing groups. These results exhibited that these plants does not contain any toxic substances.

Antibacterial, Antifungal and antioxidant activities of some medicinal plants.

The purpose of this study was to evaluate the antibacterial, antifungal and antioxidant activities of medicinal plants. The antibacterial activity of methanolic extracts of three medicinal plants (Swertia chirata, Terminalia bellerica and Zanthoxylum armatum) were tested against Gentamicin (standard drug) on eleven gram positive and seventeen gram negative bacteria by agar well method. It was revealed that seven-gram negative and six gram positive bacterial species were inhibited by these plant extracts. Minimum inhibitory concentrations (MIC) of the extracts were determined by broth micro-dilution method. The significant MIC value of Swertia chirata was 20mg/ml against Serratia marcesens, Zanthoxylum armatum was 10 mg/ml against Aeromonas hydrophila and Terminali bellerica was 20mg/ml against Acinetobacter baumanii as well as Serratia marcesens. Antifungal screening was done for methanolic extracts of these plants by agar well method with the 6 saprophytic, 5 dermatophytic and 6 yeasts. In this case Griseofulvin was used as a standard. All saprophytes and dermatophytes were showed resistance by these plants extracts except Microsporum canis, which was inhibited by Z. armatum and S. chirata extracts. The significant MIC value of Zanthoxylum armatum was 10mg/ml against Microsporum canis and Swertia chirata was 10mg/ml against Candida tropicalis. The anti-oxidant study was performed by DPPH free radical scavenging assay using ascorbic acid as a reference standard. Significant antioxidant activities were observed by Swertia chirata and Zanthoxylum armatum at concentration 200µg/ml was 70% DPPH scavenging activity (EC50=937.5µg/ml) while Terminalia bellerica showed 55.6% DPPH scavenging activity (EC50=100µg/ml). This study has shown that these plants could provide potent antibacterial compounds and may possible preventive agents in ROS related ailments.

Phytochemical, antimicrobial, insecticidal and brine shrimp lethality bioassay of the crude methanolic extract of Ajuga parviflora Benth.

Methanolic extract of medicinal herb Ajuga parviflora Benth. was evaluated for phytochemical screening (the plant extract showed the presence of aromatic compounds, carbohydrates, glycosides, tannins, alkaloids, polyphenols, quinines and dions, aminophenols, steroids/sterols, flavonoids and terpenoids), antimicrobial activities against various strains of bacteria and fungi by using disc diffusion method and insecticidal activities against red flour beetle (Tribolium castaneum), wheat weevil (Sitophilis granaries) and their larvae. The crude extract showed anti-bacterial activity against all strains with a maximum zone of inhibition of 12mm diameter against Citrobacter and Pseudomonas aurogenosa. Standard drugs Ampicillin, Gentamicin and Amoxicillin were used in parallel. The crude extract did not show antifungal activity against the tested strains of fungi even at high doses. The crude methanolic extract was also used for insecticidal activity against the two types of insects and their larva. The extract showed no significant mortality in the tested strains. For brine shrimp lethality bioassay different concentrations 10, 100 and 1000ug/ml of the medicinal herb A. parviflora methanolic extract were used. After 24 hrs the percent mortality and LD50 value was calculated through probit analysis. The LD50 value of extract was 321.42µg/mL while that of standard drug cyclophosphamide was 16.09ug/ml.

Pharmacological and biological evaluation of extracts from Gratiola officinalis L. (Scrophulariaceae).

The crude extract of Gratiola officinalis and its n-hexane, chloroform, ethyl acetate, n-butanol and aqueous fractions were subjected to biological (Brine Shrimp Bioassay, Insecticidal and Phytotoxicity/Cytotoxic) and neuropharmacological (Head dip, Open field Forced swimming test, Sodium pentothal induced sleep) activities. Results obtained in this study indicated that at high concentration dose (1000µg/ml), all test samples showed 60-95% phytotoxicity. In crude extract, n-butanol and aqueous fractions produced more than 85% phytotoxicity. While low concentration (10µg/ml) dose showed 25-28% phytotoxicity in all test samples. The crude extract was devoid of any effect against the growth of Callosbruchus analis and Tribolium castaneum and caused 10 mortality of Rhyzopertha dominica. n-Hexane, chloroform, ethylacetate, n-butanol and aqueous fractions caused 50, 30, 40, 10 and 20% mortality respectively of C. analis where as chloroform, ethyl-acetate, aqueous and crude extract, n-hexane, ethyl-acetate fractions also caused low mortality (10%) of Tribolium castaneum and Rhyzopertha dominica respectively. In cytotoxic assay at 1000µg/ml concentration, n-butanol fraction produced 36.7% and the crude extract produced 13.3% mortality of brine shrimp, its aqueous fraction was inactive at all concentrations. The results of head dip, open field, mobility time and Pentothal Na induced sleep indicated that crude extract, n-butanol and ethylacetate fractions of G. officinalis had mild sedative effect. However aqueous fraction was found to produce a significant decrease in motor activities and potentiated the duration of sleep.

Biological screening of Scrophularia nodosa extract and its fractions.

Biological screening of Scrophularia nodosa crude extract and its fractions (hexane, chloroform, ethyl-acetate, n-butanol and aqueous) was carried out on phytotoxicity, cytotoxicity, antibacterial, antifungal and analgesic activities. Crude extract and its fractions produced 50-100% phytotoxicity at 1000µ/mL concentration whereas 25-77% phytotoxicity was observed at 10µ/mL concentration. The fractions exhibited significant antibacterial and antifungal effects. The non-toxic results of this plant were recorded in Brine Shrimps Bioassay method at all concentrations. Similarly no significant insecticidal activity was observed in crude extracts and fractions. Analgesic activity results of S. nodosa in mice were found highly significant in crude extract as compared to fractions. In writhing test crude extract at 500 mg/kg showed 65.6% highest inhibitory response in mice.

Spasmolytic effects of Scrophularia nodosa extract on isolated rabbit intestine.

Scrophularia nodosa (figwort), an indigenous medicinal plant grows in moist and cultivated waste ground. It contains saponins, cardioactive glycosides, flavonoids, resin, sugar and organic acids. It is traditionally used for anti-inflammatory purpose and in skin disorders. It has diuretic and cardiac stimulant properties. The present studies were carried out on crude extract of Scrophularia nodosa and its n-hexane, chloroform, ethyl acetate, n-butanol and aqueous fractions. During phytochemical studies seven known compounds of flavonoid nature were isolated from the chloroform fraction of crude extract of S. nodosa. The structures of these compounds were elucidated by spectroscopic (UV, IR, Mass (EIMS, HREIMS) and NMR ((1)H-NMR, (13)C-NMR, DEPT, and (1)H-(1)H, COSY, HMQC, HMBC and NOESY) techniques. Compound 1 was identified as 5, 4`-hydroxy-3, 6, 7-trimethoxyflavone, compound 2 as 5-hydroxy-3,6,7,4'-tetramethoxyflavone, compound 3 as Centaurein, compound 4 as 5-hydroxy-7,8,2',3',4'-pentamethoxyflavone (Serpyllin), compound 5 as Kaempferol 7-O-α-L-rhamnopyranoside, compound 6 as sakuranetin 4'-O (6''-O-α-L-rhamnopyranosyl)-ß-D-glucopyranoside (Vitexoside) and compound 7 as Spinoside. Crude extract and its fractions were tested on isolated rabbit intestine (in vitro) for their effects. The results of crude extract and its fractions in different doses showed the decrease in normal movement of the smooth muscles of rabbit intestine (jejunum). The chloroform fraction showed maximum relaxant effect (77.37%) at 15mg/ml dose and aqueous fraction showed 38.56% spasmogenic response which was not present in the crude extract. Further study was carried out on different fractions to investigate the possible mechanism of action of S. nodosa extract. For this purpose spasmolytic effect of different fractions were compared with agonist and antagonist activities of standard drugs including adrenaline, atropine andacetylcholine (1x10(-2), 1x10(-4) and 10(-6) M conc.). It is concluded that the chemical constituents present in S. nodosa having spasmolytic action are possibly acting through muscarinic receptors.

Differential inhibitory potencies of alcoholic extract of different parts of Dryopteris chrysocoma on inflammation in mice and rats.

Phytomedicinal therapy for inflammation is not new and it is highly effective for the treatment of various inflammatory disorders. The inflammation is one of the initial parameter for most of the disorders occurring in the body. The anti-inflammatory potential can be determined by using various techniques. Dryopteris chrysocoma is a male fern commonly found in damp and moist areas of Pakistan. The study was conducted on mice and rats by inducing inflammation with subcutaneous administration of formalin and Carrageenan in hind paw. The results were compared with standard drug Aspirin administered orally in the dose of 300 mg/kg and a decrease in hind paw volume was observed. The intensity of edema was observed in mice after formalin injection and the time of disappearance of edema was observed. In rats the inhibition of inflammation by root, leaves and stem extract was 51.19%, 41.66% and 30.95% respectively after administration of formalin. Similar inhibition of inflammation produced by root, leaves and stem extracts i.e. 57%, 42% and 35% respectively in Carrageenan treated rats. Root extract showed the highly significant results at p ≤ 0.05. The comparative study explored the root extract has more potent anti-inflammatory activity than leaves extract. The stem extract has less potent anti-inflammatory action.