Assessment of Iris albicans lange as potential antimicrobial and analgesic agent.

Objective of the present work was to evaluate the presence of phytochemical constituents and pharmacological activities (antimicrobial and anti-nociceptive) of crude extract isolated from Iris albicans and its corresponding fractions. Extraction was made with methanol and extract was evaluated for the presence of different bioactive constituents, as per standard protocols. Extract and its corresponding fractions were evaluated for their antimicrobial and anti-nociceptive potential. Well diffusion method was used to evaluate the antibacterial effects while anti-nociceptive effect was studied using in-vivo models (acetic acid induced writhing, hotplate and tail immersion tests) at different dose level (100, 200 and 300 mg/kg of body weight) and compared with diclofenac sodium (dose = 10 mg/kg body weight). Results showed that the I. albicans extracts contained secondary metabolites including alkaloids, phytosterols, flavonoids, saponins, terpenoids, tannins, phenols, steroids, fixed oil, glycosides and carbohydrates. Furthermore, it was observed that different fractions in decreasing polarity order such as chloroform >n-Hexane > Ethyl acetate > Crude Methanol > Aqueous extract exhibited effective antimicrobial response against all test organisms. Results of the study showed that the extracts have significant antimicrobial and analgesic activity, providing evidence for its folklore use.

The hematological and histological studies for the hepatoprotective-like effect of the hydromethanolic extract and the fractions of Viola serpens Wall.

Traditionally, Viola serpens has been used in the treatment of several human disorders including liver diseases without any scientific evidence. As the current therapies are not very effective and face challenges of unwanted effects and patient compliance, therefore more effective and safe agents are highly needed. The current study aimed to evaluate the hepatoprotective potential of the crude extract and subsequent fractions of the whole plant in the in-vivo model using various hematological and histopathological parameters followed by an HPLC study for the identification of phenolic compounds. Rabbits (1000-1200 g) were used in the study. Paracetamol (2g) was used to induce hepatotoxicity in experimental rabbits. The plant extract was used in two doses (150 and 300 mg/kg body weights) for eight days. The hematological parameters AST, ALT and ALP values were determined along with the histopathology of the liver. Phenolic compounds were identified by high-performance liquid chromatography (HPLC) Agilent-1260 infinity from their retention time, UV spectra and available standards while quantification was done taking the percent peak area. The doses 150 and 300 mg/kg body weight seemed to be more effective. The hematological values and the histopathological slides show the hepatoprotective effect of the plant. Regeneration indicated the presence of nuclei, nuclear cleaning, prominent nucleoli, RBC's, central veins and plates of hepatocytes. The HPLC studies revealed the presence of a number of phenicol compounds. The crude extract and the subsequent fractions of the plant possess strong hepatoprotective activity, providing a scientific rationale for its uses in the treatment of liver toxicities.

Nephroprotective effect of the hydromethanolic extract and fractions of Viola serpens Wall. Histological and hematological evidence.

The current study was planned to examine the nephroprotective effect of the crude extract and its various fractions of Viola serpense Wall against paracetamol-induced toxicity in rabbits. The serum creatinine levels of all fractions, as well as the crude extract, were found to have a greater effect. The effect on urine urea by the n-hexane, ethyl acetate, n-butanol and aqueous fraction in high doses (300 mg/kg b.wt.) and crude extract and chloroform in low doses (150 mg/kg bwts.) were comparatively more effective and comparable to silymarin. The creatinine clearance of the fractions except for chloroform, aqueous at 300 mg/kg and the hydro-methanolic extracts at both doses were highly significant. The histological structures of kidneys in crude extract and chloroform-treated groups showed more improvement at the lower doses. The fractions n-hexane, ethyl acetate and n-butanolic exhibited an inverse dose relationship in the histology of the kidney. However, the aqueous fraction showed a dose-dependent nephroprotective effect. Finally, the crude extract and fractions significantly improved paracetamol-induced nephrotoxicity in rabbits.

In vivo acute toxicity, laxative and antiulcer effect of the extract of Dryopteris Ramose.

Dryopteris ramosa (D. ramosa) is one of the most traded medicinally important plants of Himalayan region. Apart from other uses, D. ramosa is traditionally also used to treat gastric ulcers and as a laxative. The present study was designed to investigate the role of methanolic crude extract of Dryopteris Ramosa (MEDR) in acute toxicity, against loperamide induced constipated mice model, antiulcer effect of methanolic extract of D. Ramosa and cholinomimetic like effect of methanolic extract of D. Ramosa. The crude extract was investigated for the presence of active compounds (secondary metabolites) such as alkaloids, flavonoids, carbohydrates, glycosides, terpenoids, phenolic compounds, saponins, and tannins following the standard methods. The antiulcer effect was investigated in mice using the ethanol induced ulcer model at various doses i.e. 50 mg/kg, 100 mg/kg and 200 mg/kg doses. Constipation was induced in the mice via loperamide (3mg/kg body weight). The control group received normal saline. Different doses of plant extracts (50, 100, 150 and 200 mg/kg body weight/day) were administered for 7 days. Various parameters like feeding characteristics, gastrointestinal transit ratio, body weight, fecal properties and the possible mechanism of action of D. Ramosa on intestinal motility were monitored. Various Phytochemicals like saponins, glycosides, flavonoids, tannins, phenols, carbohydrate, alkaloids and triterpenes were found in D. Ramosa. The acute toxicity study showed that MEDR was associated with no mortality except mild and moderate sedation at the highest tested doses (1500 and 2000 mg/kg). MEDR also showed significant antiulcer activity against ethanol-induced ulcerogenesis. The extract enhanced the intestinal motility, normalized the body weight of constipated mice and increased the fecal volume which are indications of laxative property of the herb. The 200 mg/kg body weight dose of the extract was found effective. The presence of various Phytochemicals such as flavonoids, glycosides and tannins might be responsible for the antiulcer activity of D. Ramosa. This study provides the scientific background for the folkloric use of D. Ramosa as antiulcer agent. The laxative action of the extract compares positively with Duphalac, (standard laxative drug). These findings have therefore evidence scientific background to the folkloric use of the herb as a laxative agent.

Phytochemical profiling and antioxidant potential of Daphne mucronata Royle and action against paracetamol-induced hepatotoxicity and nephrotoxicity in rabbits.

The paracetamol-induced injuries of liver and kidneys in animals are mostly used to screen out the hepato and nephroprotective effect of extract or other therapeutic agents. In the present study total phenolic and flavonoid contents, in vitro antioxidant, and in vivo hepato/nephroprotective (on paracetamol-induced intoxication in experimental rabbits) potentials of the Daphne mucronata leaves methanolic extract were determined. For the identification of possible phytochemicals, HPLC (high performance liquid chromatography) analysis was carried out and a total of eight phenolic compounds; malic acid, gallic acid, chlorogenic acid, epigallocatechin gallate, quercetin, morin, ellagic acid, and rutin were identified. D. mucronata extract at doses of 250 and 500 mg/kg body weight were given for eight days to paracetamol intoxicated rabbits and the observed results were compared with standard Silymarin. The level of liver enzymes like aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, serum triglyceride, serum cholesterol, serum bilirubin, and kidneys biomarkers like serum urea, uric acid, and creatinine, as well as lipid peroxidation malondialdehyde contents were increased while the antioxidant enzymes like reduced glutathione and total antioxidant capacity were decreased. Furthermore, histopathological analysis of the liver and kidney tissues of control and treated groups also confirmed the hepatoprotective and nephroprotective effect of the D. mucronata which was most probably due to its high antioxidant phenolic and flavonoid phytoconstituents.

Zoonotic and non-zoonotic helminths in black rats of rain-fed and irrigated areas of Swat, Khyber Pakhtunkhwa, Pakistan.

Present study was conducted to get information on helminth parasites of zoonotic importance among the black rats of district Swat, Pakistan. Two hundred and sixty nine rats were captured from agricultural ecosystem of the district using live captured traps from 2011 to 2013. Captured rats were anesthetized and surveyed for the presence of ectoparasites, then were carefully dissected for investigation of endoparsites. Helminth parasites of 8 species were identified. Presence of parasite was noticed in 23.7% of sampled rats. The infection rates of sampled rats was given in order of their infectivity as Syphacia obvelata 13(4.83%), Aspiculuris tetraptera 13(4.83%), Heterakis spumosa 12 (4.46%), Hymenolepis spp. 9(3.34%), H.diminuta 8(2.97%), Hymenolepis fusa 4(1.48%), Lutziella microacetabularae 4(1.48%) and Lutziella spp. 1 (0.37%). No significant difference (P < 0.4289) was found in prevalence of parasites among areas, crops, crop stages and sex of the host while adult rats were found more infected than sub-adults. S. obvelata and A. tetraptera were the most common species of helminths while Lutziella sp., 1 (0.37%) was found only in one host. Rattus rattus (the black rat) was regarded as the host of helminth parasites of zoonotic importance, therefore the hidden health hazards of this rodent species needed to be considered to prevent infectivity of humans. Current study was concluded that Rattus rattus harbored a wide variety of helminth parasites which shows a hidden risk to inhabitants of the region. Monitoring rats' population in settle areas and educating the local community about the risk of rat borne parasitic diseases transmission through rats appears to be absolutely essential.

Antinociceptive and Anti-Inflammatory Like Effects of Berberis baluchistanica.

BACKGROUND: Numerous therapeutic agents are in clinical practice for the treatment of inflammatory and painful conditions, but their applications have been challenged by various side /toxic effects. Therefore, new effective and safe therapies are most warrant, for which medicinal plant could be a significant alternative. Berberis baluchistanica has traditionally been used as analgesic and anti-inflammatory without any scientific background. OBJECTIVE: The current study was designed to evaluate the analgesic and anti-inflammatory like effects of extract B. baluchistanica in animal models. METHODS: For the study of antinociceptive effect, an extractof B. baluchistanica (100, 200 and 300 mg/kg i.p.), was tested in acetic acid-induced writhing and formalin tests. While for the anti-inflammatory action, carrageenan-induced paw edema, cotton pellet induced granuloma and xylene induced ear edema tests were used. RESULTS: The results showed the significant dose-dependent antinociceptive effect of extract of B. baluchistanica in acetic acid-induced writhing and formalin-induced flinching behavior tests. However, the effect was strongly antagonized by the injection of naloxone, suggesting the expression via opioidergic receptors. Similarly, strong anti-inflammatory action was illustrated in carrageenan- induced paw edema, cotton pellet induced granuloma and xylene induced ear edema tests and thus provided evidence for the versatile phytochemical nature of its phytochemical. CONCLUSION: In short, it is concluded that the extract of B. baluchistanica exhibited significant antinociceptive and anti-inflammatory effects in animal models. Further detailed studies on the efficacy and safety as well as on the phytochemical investigation are required to ascertain its clinical uses.

Involvement of selective GABA-A receptor subtypes in amelioration of cisplatin-induced neuropathic pain by 2'-chloro-6-methyl flavone (2'-Cl-6MF).

Cisplatin-induced peripheral neuropathic pain is a common adverse effect of chemotherapy. The present study evaluated the effects of 2'-chloro-6-methylflavone (2'-Cl-6MF) at recombinant α1ß2γ2L, α2ß1-3γ2L, and α3ß1-3γ2L GABA-A receptor subtypes expressed in Xenopus oocytes and subsequently evaluated its effectiveness in cisplatin-induced neuropathic pain. The results showed that 2'-Cl-6MF potentiated GABA-elicited currents at α2ß2/3γ2L and α3ß2/3γ2L GABA-A receptor subtypes. The potentiation was blocked by the co-application of flumazenil (a benzodiazepine (BDZs) site antagonist). In behavioral studies, mechanical allodynia was induced by intraplantar injection of cisplatin (40 µg/paw) in Sprague Dawley rats, and behavioral assessments were made 24 h after injection. 2'-Cl-6MF (1, 10, 30, and 100 mg/kg, i.p.), was administered 1 h before behavioral evaluation. Administration of 2'-Cl-6MF (30 and 100 mg/kg, i.p) significantly enhanced the paw withdrawal threshold and decreased mechanical allodynia. The standard drugs, gabapentin (GBP) at the dose of 70 mg/kg, and HZ 166 (16 mg/kg), i.p. also significantly enhanced the paw withdrawal threshold in mechanical allodynia. Pretreatment with pentylenetetrazole (PTZ) (15 mg/kg, i.p.) and flumazenil reversed the antinociceptive effect of 2'-Cl-6MF in mechanical allodynia indicating GABAergic mechanisms. Moreover, the binding mechanism of 2'-Cl-6MF was rationalized by in silico modeling tools. The 3D-coordinates of α2ß2γ2L and α2ß3γ2L were generated after homology modeling of the α2 subtype and 2'-Cl-6MF was at predicted binding sites of the developed models. The α2 model was compared with the α1 and α3 subunits via structural and sequence alignment. Molecular docking depicted that the compound binds efficiently at the neuromodulator binding site of the receptors. The findings of this study revealed that 2'-Cl-6MF ameliorated the manifestations of cisplatin-induced neuropathic pain in rats. Furthermore, we also conclude that GABAergic mechanisms may contribute to the antinociceptive effect of 2'-Cl-6MF. The molecular docking studies also confirm the involvement of the BDZs site of GABA-A receptors. It was observed that Ile230 of α2 stabilize the chlorophenyl ring of 2'-Cl-6MF through hydrophobic interactions, which is replaced by Val203 in α1 subunit. However, the smaller side chain of Val203 does not provide hydrophobic interaction to the compound due to high conformational flexibility of α1 subunit.

Smart nanocrystal of indomethacin: Nanonization and characterization through top down method of media milling.

Indomethacin is potent and effective drug belongs to NSAID group having low bioavailability. To address this issue the novel method is Nanosuspensions which can be achieved through bottom up and top down methods. The drug concentration, batch size and crystallinity retention are the problems associated with bottom up method consequently top down method was applied. In current project batch size of 350 ml was prepared by mixing 3.5% of Indomethacin with polymer solution. Then it was introduced into Dena⌖ having 0.2µm yttrium reinforced zirconium beads. The effect of milling time was observed for sixty minutes. Stable nanocrystals with particle size of 161nm ±1.90 with PDI of 0.229 ±0.06 were produced. The DSC and PXRD confirmed the crystallinity of created nanocrystals. The pattern of particle size reduction was initially abrupt and then gradual. The two months Stability studies at 4°C and at 25°C revealed that polymers combination (PVP-K30, HPMC-6cps, SDS) were effective in marinating the stability. The SEM and TEM studies unfastened that nanocrystals were homogenously distributed with discrete crystalline morphology. The fabricated nanocrystals demonstrated marked dissolution rate compared to the raw and marketed formulations. It is demonstrated that it is useful for industry due to high drug concentration, large batch size and retention of distinct characteristics.

Loading AKBA on surface of silver nanoparticles to improve their sedative-hypnotic and anti-inflammatory efficacies.

Aim: Acetyl-11-keto-ß-boswellic acid (AKBA) is a potent anti-inflammatory compound limited by its low water solubility and bioavailability. To load AKBA on silver nanoparticles (AgNPs) to improve bioavailability and water solubility of the compound. Materials & methods: AKBA-AgNPs were chemically synthesized and characterized by UV-Vis spectrophotometry, Fourier transform infrared spectroscopy, scanning electron microscopy and transmission electron microscopy. AKBA and AKBA-Ag were studied for their sedative-hypnotic and anti-inflammatory efficacies. Results: Pretreatment with AKBA or AKBA-Ag caused significant dose-dependent sedative-hypnotic effects at 5 and 10 mg/kg intraperitoneal. The effects of AKBA-loaded AgNPs caused pronounced changes in mice compared with those of AKBA, and the AKBA-AgNPs demonstrated anti-inflammatory effects that were superior to those of AKBA. Conclusion: The loading of AKBA on nanoparticles improved its pharmacokinetic effects, and capacity for drug delivery.

Soil-transmitted helminth infections in school children of three districts of Malakand region, Khyber Pakhtunkhwa, Pakistan.

Information on prevalence of soil-transmitted helminth (STH) infections among school children is scarce in Pakistan. This study was aimed to investigate the prevalence of soil-transmitted helminth in school children of three districts in, Khyber Pakhtunkhwa, Pakistan. A total of 300 stool samples were examined from August 2015 to August 2016 using direct smear (Normal saline and Lugol's Iodine solution) and the concentration methods. One hundred and eighty seven (62.3%) pupils were found infected with soil-transmitted helminths. One hundred and forty five (77.5%) were infected with single parasite and forty two (22.4%) with multiple infections. Ascaris lumbricoides 125 (66.4%), Trichuris trichura 50 (26.5%) and Ancylostoma duedenale 13 (6.91%) were detected. The children above 8 years in age were more parasitized than below 8 years (p=0.7832; P>0.05). Males were found more parasitized than females (p=0.9315; P>0.05). Children in lower Dir district were found more infected followed by Swat and upper Dir (P< 0.0001; p<0.05). No significant relationship was found among the examined and that of infected children for ages and sex in all the districts. Malakand division is an area with poor hygiene located in temperate zone near the border of Afghanistan and China. The prevalence of reported nematode parasites here compared with the same studies is unexpectedly high. These types of studies should continue time to time to know the hazardous nature of such parasitic infections for the betterment of the human health.

Enhanced dissolution rate of Ketoprofen by fabricating into smart nanocrystals.

The low bioavailability of Ketoprofen is associated with its hydrophobic nature that can be solved by nanonization. For this purpose, a polymeric solution with drug concentration of 3.5% w/w was formulated. The produced solution was milled for 60 minutes in DENA® mill which contains 0.2µ m yttrium reinforced zirconium beads. The Physicochemical properties, characterization including stability studies of the prepared nanoparticles were carried out using pharmacopeial techniques of zeta potential, PXRD, DSC, SEM and TEM. Results suggest that stable crystalline nanocrystals with a size of 169±1.98nm with PDI of 0.194±0.04 and zeta potential of -22.0±2.25mV were produced. Moreover, enhances in-vitro release rate of 78.6% for the processed Ketoprofen was achieved in first 5 min as compared to raw form and marketed drug which released only 22.9% and 33.1% of drug respectively. The 60 days stability studies at 4oC & 25°C revealed that polymers PVP-K30-HPMC-6cps-SDS were effective in stabilizing the nanocrystals. Comparatively stable ketoprofen nanocrystals were successfully produced by DENA® mill with marked enhanced dissolution rate. It proved a useful for commercialization technique due to high drug concentration and retention of distinct characteristics at large scale.

In vitro antioxidant and hepatoprotective activities of Paeonia emodi (Wall.) rhizome methanol extract and its phenolic compounds rich fractions.

The present study aimed to quantify the total phenolic content in Paeonia emodi rhizome methanol extract and its fractions and then evaluate the in vitro antioxidant and hepatoprotective activities of fractions rich in phenolic compounds. Maximum quantity of total phenolic content was observed in butanol (112.08±5.5 mg GAE/g dw) and chloroform fraction (107.0±3.5 mg GAE/g dw) followed by methanol extract (94.2±4.4 mg GAE/g dw), aqueous fraction (92.9±2.5 mg GAE/g dw), ethyl acetate (62.3±8.3 mg GAE/g dw) and n-hexane fraction (51.6±7.2 mg GAE/g dw). The fractions rich in total phenolic content were evaluated for in vitro antioxidant activity based on 2, 2-diphenyl-1-picryl-hydrazyl (DPPH) scavenging assay. The butanol and chloroform fraction showed significantly (P<0.05) higher radical scavenging activity with IC50 values of 6.5 and 7.05±2.5 ppm respectively. Positive correlation (R square=0.95) was observed between total phenolic content and in vitro antioxidant activity. The fractions rich in phenolic compounds were also evaluated for their hepatoprotective activity in paracetamol intoxicated mice. Five days oral administration of these fractions at a dose of 300 mg/kg body weight restored the serum ALT, AST and ALP levels of paracetamol intoxicated mice to normal level. From the results of the present research it was concluded that the butanol and chloroform fractions of P. emodi rhizome methanol extract are rich in phenolic compounds and strong antioxidant and effective in attenuation of hepatotoxicity.

Antimicrobial activities of rhizomes of Polygonatum verticillatum: attributed to its total flavonoidal and phenolic contents.

The current study was undertaken to evaluate the rhizomes of Polygonatum verticillatum against various pathogenic bacteria and fungi. Broad spectrum antibacterial activity was demonstrated by the crude extract of the plant and its subsequent solvent fractions; predominantly against Gram-negative bacteria. MICs of the extracts against Escherchia coli, Salmonella typhi and Shigella flexeneri were in the range of 1.5-40 µg/ml, 03-06 µg/ml and 03-40 µg/ml, respectively. The only sensitive Gram-positive bacterium was Staphylococcus aureus with MICs in the range of 75-80 µg/ml. The fungicidal activity was limited to Microspoum canis and Fusarium solani and the MICs were in the range of 350-360 µg/ml and 190-290 µg/ml respectively. The various fractions of rhizomes contained significant concentration of total flavonoidal and total phenolic contents that could be responsible for the current findings.