In-vitro antigout potential of Alstonia scholaris flower, characterization and prospective ligand-receptor interaction of bioactive lead compound.

Gout is an arthropathic and inflammatory disease. The prevalence and incidence of such disease has risen in last decades. It is associated with life style thus it could be recognize as life style diseases. In the present study, the flower extract of Alstonia scholaris Linn R.Br., Flower was initially subjected to extraction, isolation which leads to purification of pure eight compounds. All these compounds were identified using various spectroscopic techniques. In-vitro Xanthine oxidase inhibition activity was performed to determine the antigout potential of lead compounds. Compound 8 showed significant activity among all i.e. 14.7 ± 0.43 as compare to standard allopurinol 6.77 ± 0.26. Accordingly, in-silico studies using Autodock vina 4 showed the ligand-protein interaction of luteolin with 3AX7. The docking simulations showed significant binding pocket sites of respective proteins 3AX7 with the least binding energy -10.2 kcal/mol. Consequently, molecular docking simulations for 100ns indicated robust evidence with their conformational structural interaction which serve as active sites for Lead compound. Principal Component Analysis indicated first three PCs capture 23.8%, 39%, and 49% of structural variance in protein. Therefore compound 8 could be consider for potential drug design and development in gout therapy.

Urease inhibition and DPPH radical scavenging potential of phytoconstituent from Alstonia scholaris and molecular docking interactions of bioactive luteolin with target proteins.

A polyphenolic flavone Luteolin (3',4',5,7-tetrahydroxyflavone) is found in various plants and is traditionally used in Chinese medicine. It is obtained from Alstonia scholaris (L.) R.Br Flower belonging to the family Apocynaceae while investigation. Various studies have been demonstrated the antioxidant or antiulcer potential of luteolin from different plant sources. In the present investigation the antioxidant or antiulcer effect of the Luteolin has been carried out using molecular docking simulations. The objective of this study was to analyze the antioxidant and antiulcer potential of luteolin obtained during isolation. The in vitro biological evaluation has been supported by the in silico studies using Autodock vina 4 shows the ligand-protein interaction of lute olin with 1HD2, 4GY7 and 3O1Q. Luteolin showed significant DPPH scavenging and urease inhibition activity i.e., 23.4 ± 0.87, 6.21±0.45 IC50 (uM) respectively as compared to the standard BHA and thiourea 44.2±0.45, 22.4±0.29 IC50 (uM) respectively. The docking simulations showed significant binding pocket sites with the respective proteins1HD2, 4GY7 and 3O1Q with the least binding energy -6.8, -8.0 and -8.2 kcal/mol respectively. Thus, Strong evidence has been presented with their confirmation structural interaction via molecular docking with proteins that serve as binding sites for available Luteolin molecule. The findings justify the application of the compound as a novel antioxidant and antiulcer agent.

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).

Attenuation of cisplatin-induced acute kidney injury by N-(2-Hydroxyphenyl) acetamide and its gold conjugated nano-formulations in mice.

The attenuation of cisplatin-induced acute kidney injury (AKI) in mice by N-(2-hydroxyphenyl) acetamide (NA-2) and NA-2-conjugated gold nanoparticles (NA2-AuNPs) was investigated. Male BALB/c mice (n = 54) were divided into nine groups having six animals in each group. Animals in groups 3-9 were pre-treated for 5 days with test compounds, whereas, animals in group 1 and 2 received normal saline. On day 4, animals in groups 2, 3, 4, 5, 6 and 9 were given single intra-peritoneal injection of CP at the dose of 5 mg/kg. After 72 hours of CP injection, all animals were sacrificed. Blood was collected for serum urea and creatinine estimation, and kidneys were harvested for histo-pathological examinations and qPCR studies for nuclear factor-κB p50, (NFκB) ; inducible nitric oxide synthase (iNOS); hemeoxygenase-1 (HO-1); and interleukin-6 (IL-6).NA-2 and NA2-AuNPs was observed to decrease the serum urea and creatinine levels. Both the test compounds reduced kidney injury damage score and improved histological architecture in the treated animals in dose dependent manner. Furthermore, the mRNA expressions of NFkB p50, iNOS and IL-6 genes were down-regulated, and HO-1 gene was up-regulated in the animals treated with the test compounds. It is concluded that NA-2 and NA2-AuNPs attenuates CP-induced AKI in mice models through anti-inflammatory and anti-oxidant mechanisms.

Light transmission properties of pharmaceutical liquid bottles and evaluation of their photoprotective efficacy.

The light sensitive pharmaceutical dosage forms are well protected from light by packing in light protective bottles especially the colored glass and plastic bottles. In the present study the transmission characteristics of transparent glass bottle, amber glass bottle, polyvinyl chloride amber plastic bottle (PVC) and low density polyethylene semi-opaque plastic bottles (LDPE) (empty and drug filled) have been evaluated and the data compared for compliance with Pharmacopoeial limits of percentage transmission. The variations in thickness affect the amount of light transmitted through the bottles. For an average thickness, the transmission of bottles was not uniform indicated the effect of manufacturing variables on the transmission of light. The drug filled bottles showed an increase in light transmission probably as a result of interaction between drug and bottle components. The leaching of any coloring agents from glass bottles or the pigments from plastic bottles into the solution during storage appeared to increase the transmission of light which could be detrimental to photosensitive drugs in a formulation. The light protective efficacy of bottles was in the order: Semi-opaque plastic (LDPE) > amber plastic (PVC) > amber glass. The photoprotection of aqueous solution of riboflavin as a model compound in these bottles has been studied and its shelf-lives and stability ratio were determined.

Efficiency of proteolytic enzymes in treating lumbar spine osteoarthritis (low back pain) patients and its effects on liver and kidney enzymes.

Lumbar spine osteoarthritis with 40-85% prevalence, degeneration of spine with remarkable narrowing of disc space and osteophytes formation trigger pain in lower back. Pain in lower portion of back is now considered to be the second most commonly treated health issue in primary health care setups. This pain causes disability, functional loss and job absentees. Commonly pain is managed pharmacologically by NSAIDS but resulted in severe gastric side effects. The purpose of this trial was to appraise the properties of bromelain and papain, the vegetal proteolytic enzymes, in comparison with standard drug on LBP patients. Forty men and women with lumbar spine osteoarthritis were recruited and divided into group 1, received aceclofenac 100mg tablet b.i.d as standard treatment, group 2, patients treated with aceclofenac 100 mg tablet b.i.d and enzyme supplements 250 mg b.i.d for 6 weeks. All the participants were evaluated for their body mass index, vital signs and liver/kidney enzymes before and after treatment. Moreover intensity of pain were also measured through visual analogue scale (VAS) and oswestry low back pain questionnaire (ODI) before treatment (0 week), 3rd week and 6th week of treatment. The enzyme group patients showed significantly diminished pain scores VAS from 7.10±1.29 to 5.85±1.531*** (P=0.001), ODI score from 56.2±8.70 to 51.6±8.125*** (P=0.000), significantly diminished enzymes; ALP from 210.00±55.24 to 196.90±51.02 (P=0.054*) and serum creatinine from 0.97±0.153 to 0.87±0.139 (P=0.035*) and improved quality of life. Hence, this study suggested that the enzyme supplements for 6 weeks have prolonged beneficial carry-over effects in comparison to standard treatment without producing any change in BMI (P>0.05) and vital signs (P> 0.05).

The novel effect of propranolol in glibenclamide induced hepatotoxicity.

Glibenclamide (GBC) has been associated with hepatotoxicity in humans. This study conducted on rabbits to evaluate the hepatotoxicity of GBC alone and in combination therapy with propranolol (PPL). Liver enzymes like alanine transaminase (ALT), alkaline phosphatase (ALP) and gamma-glutamyltransferase (γ-GT) and bilirubin (BRB) are used to evaluate hepatotoxicity associated with GBC. Histological findings, micrometry and scanning electron microscopy (SEM) used to find hepatotoxicity by GBC and with PPL. GBC caused significant elevation of liver functions as compared to control (p<0.005). PPL reduced the level of serum ALT, ALP, γGT and BRB when administered with GBC (p<0.005). The results prevailed that there is a significant change in hepatic cells structure and significant change in its diameter of nucleus (p<0.05). The necrosis and granuloma with decreased in number of hepatic cells were observed in GBC treated rabbits. However, the combination of GBC with PPL has shown healthy and nearly similar structure as that of controlled group and confirmed by SEM microscopy. PPL reduced the blood flow to hepatic portal system and thus, avoid the noxious substances to liver. It is affirmed that the use of PPL offered beneficial effect on hepatotoxic drugs.

Storage of vaccines in different health centers and pharmacies at Karachi, Pakistan: The handling errors.

The fundamental human desire is prevention from the infectious disease. Vaccine is the basic element to fulfill this human craving. The proper handling of vaccine can minimize the degradation. The main aim of this study is to find out the knowledge of respondents and storage condition and handling of marketed vaccines in different private clinics, hospitals and community pharmacies in Karachi, Pakistan. The present study revealed that ~77% have been allowed to visit their vaccine storage compartment. The cumulative knowledge of respondents regarding recommended temperature, shake test, thermal, freezing and photo degradation were 49.34%, 24.01%, 95.78%, 54.35% and 47.23% respectively. The major mishandling like the medicines, food and drinks and even laboratory specimen have been noticed 76.3%, 61.62% and 5.35% respectively. Moreover, water filled bottles on upper, lower and door shelves and ice packs or ice trays 3.43% and 14.18%, respectively, have been found in refrigerators and freezers, respectively. No expired vaccines have been found. The major consequence, in the study has been found to be mishandling by the absence of non-professional personnel. Either doctors or pharmacist (health associated professionals) as respondents were nearly all aware about the consequences of the mishandling of vaccines and have the knowledge about the recommended temperature, thermal, photo degradation, and freezing hazards of vaccines. The world-renowned immunization authorities have strongly recommended national/local immunization programs. On the basis of the present study, it is recommended to train the staff and prepare a national guideline for the storage and handling of vaccines, and forcefully implements the law to adopt the particular guideline.

In vitro antibacterial susceptibility of different brands of oral levofloxacin 250 mg tablet against Staphylococcus aureus and Escherichia coli.

Antibiotics are not only used in morbidity but also help in prevention of infection. The irrational use of broad spectrum antibiotics is now increasing the resistance against pathogens. This present study has been carried out to evaluate the in-vitro antibacterial effect of levofloxacin against clinical isolates. According to Clinical and Laboratory Standards Institute (CLSI) guidelines, minimum inhibitory concentrations 90% (MIC90) of the levofloxacin tested were evaluated by an agar dilution method. Total 63 clinical isolates Staphylococcus aureus (n=34) and Escherichia coli (n=29) were collected from different hospitals at Karachi and were evaluated MIC90 of eleven different brands of levofloxacin tablet (250 mg). Levofloxacin (Reference) was tested against E.coli standard (American Type Culture Collection) (ATCC=25922) with (MIC90; 0.03µg/ml) and compared with different eleven brands of levofloxacin tablets 250mg (MIC90; 0.5µg/ml -16.0µg/ml). Levofloxacin (Reference) sensitivity against S. aureus standard (ATCC=25923) is (MIC90; 0.12µg/ml) and similarly when it was compared with same levofloxacin tablets (MIC90; 0.5-16.0µg/ml). It has been concluded by the present study, a large number of strains of bacteria have shown better bactericidal action of different brands of levofloxacin and nearly all commercialized drugs were appropriate for therapeutic use.

Effect of solvent polarity on the extraction of components of pharmaceutical plastic containers.

A study of the extraction of polymeric material and dyes from the pharmaceutical plastic containers using various organic solvents was conducted to evaluate the effect of polarity on the extraction process. The plastic containers used included semi-opaque, opaque, transparent and amber colored and the solvent used were acetonitrile, methanol, ethanol, acetone, dichloroethane, chloroform and water. The determination of extractable material was carried out by gravimetric and spectrometric methods. The yield of extractable materials from containers in 60 h was 0.10-1.29% (w/w) and the first-order rate constant (kobs) for the extraction of polymeric material ranged from 0.52-1.50 × 10-3 min-1 and for the dyes 6.43- 6.74 x10-3min-1. The values of (kobs) were found to be an inverse function of solvent dielectric constant and decreased linearly with the solvent acceptor number. The extractable polymeric materials exhibited absorption in the 200-400 nm region and the dyes in the 300-500nm region. The rates of extraction of polymeric material and dyes from plastic containers were dependent on the solvent dielectric constant. The solvents of low polarity were more effective in the extraction of material indicating that the extracted material were of low polarity or have non-polar character. The dyes were soluble in acetone and chloroform. No plastic material was found to be extracted from the containers in aqueous solution.

Formulation and stabilization of norfloxacin in liposomal preparations.

A number of liposomal preparations of norfloxacin (NF) containing variable concentrations of phosphatidylcholine (PC) (10.8-16.2mM) have been formulated and an entrapment of NF to the extent of 41.7-56.2% was achieved. The values of apparent first-order rate constants (kobs) for the photodegradation of NF in liposomes (pH7.4) lie in the range of 1.05-2.40×10(-3)min(-1) compared to a value of 8.13×10(-3)min(-1) for the photodegradation of NF in aqueous solution (pH7.4). The values of kobs are a linear function of PC concentration indicating an interaction of PC and NF during the reaction. The second-order rate constant for the photochemical interaction of PC and NF has been determined as 8.92×10(-2)M(-1)min(-1). Fluorescence measurements on NF in liposomes indicate a decrease in fluorescence with an increase in PC concentration as a result of formation of NF(-) species which exhibits poor fluorescence. Dynamic light scattering has shown an increase in the size of NF encapsulated liposomes with an increase in PC concentration. The stabilization of NF in liposomes is achieved by the formation of a charge-transfer complex between NF and PC.

Formulation and stabilization of riboflavin in liposomal preparations.

A study of the formulation of liposomal preparations of riboflavin (RF) with compositional variations in the content of phosphatidylcholine (PC) and their entrapment efficiency (26-42%) have been conducted. Light transmission characteristics of the liposomal preparations have been determined to evaluate their effect on the amount of light passing through the system to initiate a photochemical reaction. Dynamic light scattering (DLS) and atomic force microscopy (AFM) have been used to study the physical characteristics of liposomes. The liposomal preparations of RF have been subjected to photolysis using visible light and the apparent first- order rate constant, kobs, for the degradation of RF have been determined. The values of kobs (1.73-2.29×10(-3)min(-1)) have been found to decrease linearly with an increase in PC concentration in the range of 12.15 to 14.85 mM. Thus, an increase in PC concentration of liposomes leads to an increase in the stability of RF. RF and its main photoproduct, lumichrome (LC), formed in liposomes have been assayed by a two-component spectrometric method at 356 and 445 nm using an irrelevant absorption correction to compensate for the interference of liposomal components. The fluorescence measurements of RF in liposomes indicate excited singlet state quenching and the formation of a charge-transfer complex between RF and PC. It results in electron transfer from PC to RF to cause photoreduction and stabilization of RF.

In vitro bactericidal activity of cefepime and cefpirome against clinical isolates at Karachi.

Antibiotics not only support to alleviate the infections but also facilitate to avert the multiplication of microbes. Due to the irrational use of antibiotics, the resistance of antibiotics has been augmented which results may increase in morbidity and mortality with the span of time. World renowned regulatory bodies like Food and Drug Administration (FDA), Center of Disease Control and Prevention (CDC), and World Health Organization (WHO) vigorously advocate the surveillance of the resistance of antibiotics. During the present study by Kirby-Bauer disk diffusion method 141 clinical isolates of Staphylococcus aureus (n=47, 33.34%), Escherichia coli (n=54, 38.3%), Proteus species (n=26, 18.4%), and Klebsiella pneumoniae (n=14, 9.92%) are evaluated against cefepime and cefpirome which comes of fourth generation cephalosporin. It has been found that cefpirome has better bactericidal activity than cefepime against E. coli and K. pneumoniae while cefepime has been possessed better antibacterial activity against S. aureus and Proteus species which were isolated from respiratory tract infections, blood stream infection, intra-abdominal and urinary tract infections, and skin and soft tissue infections. K. pneumoniae, E. coli, Proteus species, and S. aureus were 34.8%, 26.3%, 11.3%, and 37.7% resistance against cefepime respectively. S. aureus, E. coli, K. pneumoniae, Proteus species has shown 41.4%, 21.7%, 17.6%, and 8.9% resistance against cefpirome correspondingly.

Effect of acetate and carbonate buffers on the photolysis of riboflavin in aqueous solution: a kinetic study.

The photolysis of riboflavin (RF) in the presence of acetate buffer (pH 3.8-5.6) and carbonate buffer (pH 9.2-10.8) has been studied using a multicomponent spectrophotometric method for the simultaneous assay of RF and its photoproducts. Acetate and carbonate buffers have been found to catalyze the photolysis reaction of RF. The apparent first-order rate constants for the acetate-catalyzed reaction range from 0.20 to 2.86 × 10(-4) s(-1) and for the carbonate-catalyzed reaction from 3.33 to 15.89 × 10(-4) s(-1). The second-order rate constants for the interaction of RF with the acetate and the carbonate ions range from 2.04 to 4.33 × 10(-4) M(-1) s(-1) and from 3.71 to 11.80 × 10(-4) M(-1) s(-1), respectively. The k-pH profile for the acetate-catalyzed reaction is bell shaped and for the carbonate-catalyzed reaction a steep curve. Both HCO3(-) and CO3(2-) ions are involved in the catalysis of the photolysis reaction in alkaline solution. The rate constants for the HCO3(-) and CO3(2-) ions catalyzed reactions are 0.72 and 1.38 × 10(-3) M(-1) s(-1), respectively, indicating a major role of CO3(2-) ions in the catalysis reaction. The loss of RF fluorescence in acetate buffer suggests an interaction between RF and acetate ions to promote the photolysis reaction. The optimum stability of RF solutions is observed in the pH range 5-6, which is suitable for pharmaceutical preparations.

Resistance pattern of clinical isolates involved in surgical site infections.

Wound infections due to the incursion of microbes need to be averted or to heal the wounds by antibiotics. Antibiotics are not only aid in cure of infections but also help to prevent the flourishing and production of one or more species of microorganism, resultant in purulent discharge. This current study was carried out to evaluate the resistance pattern of clinical isolates from surgical site infections by the Kirby Bauer disc diffusion method. A total of 257 clinical isolates were collected from different hospitals in Karachi and evaluated by using fifteen antibiotics belonging to different groups. Staphylococcus aureus (n=87), Escherichia coli (n=76), Pseudomonas aeruginosa (n=56), Proteus (n=21) and Klebsiella (n=17) species are the most common clinical isolates of surgical site infections. Among the semi-synthetic penicillins, ampicillin was found to be resistant to nearly all clinical isolates but amoxicillin was moderately sensitive to S. aureus. Combinations of semi-synthetic penicillins are more sensitive than the penicillin alone. Co-amoxiclave exhibits superior sensitivity to all the surgical infection isolates except Pseudomonas aeruginosa which showed 68.75% resistance. Pseudomonas aeruginosa was highly resistant to cephalosporin except ceftraixone which showed 21.88% resistance. S. aureus was slightly responsive to cefazolin, cephradine, cefaclor, ceftizoxime, cefuroxime and ceftriaxone. E. coli, Gram-negative clinical isolate was showed 25% and 31.25% resistance to ceftriaxone and cefuroxime. In the Klebsiella species, 71.42% and 64.29% resistance to cefazolin and cefuroxime respectively, was observed. Aminoglycosides such as gentamycin and tobramycin were found to be more susceptible to all the clinical isolates. Quinolones like ofloxacin and enoxacin were showed good sensitivity to nearly all the clinical isolates.On the basis of the present study, it is recommended to adopt a rational use of antibiotics in prophylaxis and the utilization of a coordinated scheme of surgical wound inspections.