Biosynthesized Silver Nanoparticles Using Alnus nitida Leaf Extract as a Potential Antioxidant and Anticancer Agent.

Biogenic synthesis of silver nanoparticles (AgNPs) using plant extracts is gaining attention as a substitute to the conventional physical and chemical synthesis methods. This study reports a facile, cost-effective, and ecofriendly synthesis of AgNPs using leaf extract of Alnus nitida (A. nitida) and their antioxidant and antiproliferative activities. The biosynthesized AgNPs were characterized using various analytical techniques including UV-visible spectroscopy, energy-dispersive spectrometry, scanning electron microscopy (SEM), Fourier transform infrared (FTIR), X-ray diffraction (XRD), and dynamic light scattering. The antioxidant and cytotoxic potential of the extract and AgNPs was evaluated using different in vitro models. The UV-vis analysis revealed a surface plasmon resonance peak of 400 nm corresponding to the synthesis of AgNPs. SEM analysis confirmed the formation of heterogeneously dispersed particles of nano size, while the XRD and FTIR spectra confirmed the crystallinity and existence of different functional groups that helped in capping and stability of AgNPs. The antioxidant activity of AgNPs and extract, studied by 1,1-diphenyl 2-picryl hydrazyl (DPPH), fluorescence recovery after photobleaching (FRAP), 2, 2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and H2O2 scavenging assays, showed a dose-dependent effect. The AgNPs at 1000 µg/mL significantly scavenged DPPH, FRAP, ABTS, and H2O2 by 66.45, 74.65, 78.81, and 72.56% with an average IC50 value of 33.31, 18.50, 16.46, and 15.65 µg/mL, respectively. The cytotoxic potential investigated by MTT assay revealed promising antiproliferative effects against different cancer cell lines. The IC50 values of AgNPs on MDA-MB-231, A549, and Hep-G2 cells were 14.88, 3.6, and 5.38 µg/mL, respectively. The results showed that AgNPs were more effective against lung and hepatocellular carcinoma. The selectivity index showed that AgNPs remained highly selective in retarding the growth of A549 and Hep-G2 cells as compared to normal cell lines HPAEpiC and HRPTEpiC. Overall, this study showed that biosynthesized AgNPs were associated with considerable antioxidant and cytotoxic effects. Our work suggests that A. nitida-mediated AgNPs should be evaluated further in order to develop safe and effective formulations for the treatment of different degenerative diseases.

The effect of gender and ABCB1 gene polymorphism on the pharmacokinetics of azithromycin in healthy male and female Pakistani subjects.

In the current study, the possible outcome of gender difference and genotypic polymorphism of the ABCB1 gene encoding P-glycoprotein on the pharmacokinetics of azithromycin has been evaluated. An open-label, comparative pharmacokinetic study was done in healthy Pakistani volunteers (females (n = 8) and males (n = 8)). They were administered a single 500 mg oral dose of azithromycin. Blood samples (≈5 mL) were collected in heparinized tubes and the HPLC/MS/MS method was used to determine azithromycin plasma levels. ABCB1 polymorphism (single nucleotide polymorphisms) at C3435T, G26SST was performed using the RFLP-PCR method. The Student t test was applied to compare pharmacokinetic parameters of azithromycin between male and female human subjects (at 95% CI) using GraphPad Prism-8. A significant difference was observed in pharmacokinetic parameters between males and females, as Cmax in males (230 ± 80.2 ng/mL) was significantly higher than in females (224.9 ± 75.5 ng/mL), while [Formula: see text] was also significantly higher (p < 0.05) in males (2102 ± 200.3 ng·h-1·mL-1) compared to females (1825.7 ± 225.4 ng·h-1·mL-1). There was a significant variation in Cmax and AUC in three ABCB1 genotyping groups as well. Gender difference and ABCB1 gene polymorphisms have a significant impact on the pharmacokinetics of azithromycin, as they contribute to interindividual variability in therapeutic response.

Evaluation and assessment of prescribing patterns in elderly patients using two explicit criteria based screening tools: (The PRISCUS list and STOPP/START criteria).

BACKGROUND & OBJECTIVES: Rational prescribing can prevent medication errors and the associated harm, especially in old age patients, as they are being frequently prescribed with drugs for various ailments. Moreover, polypharmacy is a common practice in them. Therefore, a significant threat of potential drug interactions and adverse effects exist. Current study focuses on assessment of Potentially Inappropriate Medication (PIM) in medication prescribed to old age patients. METHODS: It was a forty days, descriptive and observational study conducted from August 15th 2017 to September 25th 2017 in which prescriptions given to elderly patients were reimbursed for collecting various sets of information. In order to assess PIMs (in Pakistani Set-up), STOPP/START addition 2008 (including examples of misprescribing, overprescribing and under prescribing) and the PRISCUS list (misprescribing and overprescribing) was used. Statistical analysis of results was performed using SPSS version 20. RESULTS: One hundred forty six cases of PIMs (including incorrect prescribing, overprescribing and under prescribing) were detected. It included incorrect prescriptions 104, under prescription 28 and over prescriptions 14. NSAIDS accounted for most incorrect prescriptions followed by benzodiazepines. Mostly NSAIDS were used for myalgia, backache and rheumatoid disorders. CONCLUSION: Current findings highlighted Potentially Inappropriate Prescribing (PIP), particularly of NSAIDs and under prescribing of statins in cardiovascular diseases. Study findings suggest introducing pertinent interventions at the stages involved in prescribing, prescription review and its follow up to reduce the PIP and PIMs.

Comparative pharmacokinetics of Omeprazole and its metabolites in poor and extensive metabolizer Pakistani healthy volunteers and a review of different studies.

This study was designed to evaluate a comparative single dose (40mg) pharmacokinetics (PK) of Omeprazole (OMP) and its two metabolites, 5-hydroxy Omeprazole (5-OH-OMP) and Omeprazole sulphone (OMP-S) in poor (PM) and extensive (EM) metabolizer Pakistani healthy adult volunteers. The frequency of CYP2C19 and CYP3A4 varies widely in different populations. The present study was conducted to evaluate the PK of OMP and its two metabolites in Pakistani population and to review different studies conducted after administration of single dose of OMP. Twenty two subjects were enrolled in this study and divided into two groups. The CYP2C19 phenotyping was evaluated by the metabolic ratio of OMP to 5-OH-OMP. It was a single dose, open label study and the blood samples from subjects were collected at different time intervals until 24 hours. The PK parameters were calculated using the PK-summit software. The metabolic ratio of area under the plasma concentration-time curve AUCOMP/5-OH-OMP was 1.86 ± 0.572 and13.84 ± 2.504 for EM and PM, respectively; maximum plasma concentration (Cmax) of OMP was increased by two folds for PM while the AUC∞ was increased by 3 folds; the Cmax and AUC∞ of 5-OH-OMP decreased for PM by 2 folds while there was 3 fold increase observed in the Cmax and AUC∞ of OMP-S. The PK of OMP and its metabolites in different populations were also discussed, and issues regarding CYP2C19 and CYP3A4 genotyping were also extensively reviewed. In EM of CYP2C19 the concentration of 5-OH-OMP is higher while that of OMP-S is lower. This study as well as reported studies reveals that in PM of CYP2C19 more drugs are available for CYP3A4 to be metabolized. A correlation between CYP2C19 EM and PM activity with CYP3A4 needs to be established.

In vitro Studies on Cytotoxic, DNA Protecting, Antibiofilm and Antibacterial Effects of Biogenic Silver Nanoparticles Prepared with Bergenia ciliata Rhizome Extract.

BACKGROUND: Many health hazardous diseases are caused by clinical pathogens. Drugresistant microbes are one of the major health problems in the world. To overcome the effect of infectious diseases new antimicrobial agent from nature has been explored which is environmentally friendly, less costly and more effective for the development of next-generation drugs. Bergenia ciliata and silver nitrate both have medicinal properties. OBJECTIVES: The aim of the current research was to evaluate the cytotoxic, and antibacterial effect of green synthesized nanoparticles using Bergenia ciliata rhizome against clinical bacterial pathogens. METHODS: Extract of Bergenia ciliata was prepared by maceration technique. Silver nanoparticles were synthesized using Bergenia ciliata rhizome extract. Synthesized silver nanoparticles were confirmed by UV-vis spectrophotometer, Scanning electron microscope (SEM) and Fourier Transform Infrared Spectroscopy (FTIR). The antibacterial, anti-biofilm, cell proliferation inhibition, DNA protection, brine shrimp lethality effects of synthesized nanoparticles were investigated. RESULTS: UV-vis spectrophotometer indicated the prelaminar synthesis of silver nanoparticles at 400 nm. The spherical shape of synthesized nanoparticles with 35 nm size was confirmed using SEM. Greatest zone of inhibition (6.0 ± 0.0 mm to 8.3 ± 0.57 mm) was recorded against all tested pathogens compared with the B. ciliata aqueous extract. Anti-biofilm analysis and MTT assay supported the results of the antibacterial activity. Silver nanoparticles protect the DNA degradation. CONCLUSION: Green synthesized nanoparticles had potent antibacterial activity and may provide a basis for the development of the new antibacterial drug.

Effect of clopidogrel on the hydroxylation and sulfoxidation of omeprazole: A single dose study in healthy human volunteers.

Based upon the known potential interaction between omeprazole (OMP) and clopidogrel (CLOP), the current study was designed to evaluate the effect of CLOP on disposition of OMP and its two major metabolites, 5-hydroxyomeprazole (5-OH-OMP) and omeprazole sulfone (OMP-S) in healthy clinical subjects. A randomized, open label, 2-period, crossover study was designed. Twelve volunteers were selected, of whom eight were extensive metabolizers (EM) of CYP2C19 and 4 were poor metabolizers (PM). They received single dose of OMP either alone or in combination with CLOP (single dose) and samples were collected periodically to calculate various pharmacokinetic parameters. Changes in most of the pharmacokinetic parameters of OMP, 5-OH-OMP and OMP-S were insignificant (P ˃ 0.05) both in EM and PM except for the maximum concentration (Cmax) of 5-OH-OMP and OMP-S in EM. The OMP Cmax and AUC0-∞ was increased both in EM and PM after concomitant administration of OMP with CLOP. The 5-OH-OMP Cmax was decreased in both EM and PM, demonstrating that CLOP inhibits hydroxylation of OMP. The OMP-S Cmax and AUC0-∞ were increased both in EM and PM showing that CLOP may induce sulfoxidation of OMP. It was concluded that CLOP may inhibit hydroxylation of OMP to a greater extent in EM than in PM, leading to higher OMP Cmax and AUC0-∞. Furthermore, the sulfoxidation of OMP may also be induced by CLOP. So, it is suggested that both these drugs should be carefully prescribed together to avoid any harm to the patients. (Application number13/EC/Pharm. Ref number 12/Pharm).

Assessment of antibiotic self-medication practice among public in the northwestern region of Pakistan.

BACKGROUND: Self-medication with antibiotics is a common practice, which may lead to the development of antimicrobial resistance (AMR)-a major health concern worldwide. The most common reason for the development of AMR is a lack of education and regulatory policies and the lack of community pharmacists. OBJECTIVE: To assess various factors that lead to self-medication with antibiotics, which might cause AMR and hinder effective healthcare. METHODS: A cross-sectional study was carried out using a predesigned questionnaire to collect data from 800 respondents. The respondents were selected by simple random sampling during November 2014 to January 2015 from different regions of Khyber Pakhtunkhwa (KPK), Pakistan. Only properly completed questionnaires were assessed for different variables. The collected data were analysed using SPSS V.16. RESULTS: 527 people completed and returned the questionnaire-a response rate of 66%. Self-medication with antibiotics was reported by 135 participants (26%), with a higher prevalence of men than women (48% vs 38%, respectively). The main reason for self-medication was previous experience with the same antibiotic (68%). The most commonly used antibiotics were amoxicillin-clavulanate (40%) and major indications for self-medication were sore throat (29%) and flu (24%). Of the 527 respondents, only 104 (20%) were aware of AMR. CONCLUSIONS: This study is the first to evaluate self-medication with antibiotics in KPK, Pakistan. In view of the high prevalence of self-medication, introduction of a public health policy through drug regulatory authorities, public awareness programmes/campaigns, patient education about AMR and appropriate use of antibiotics are critically required. The role of community pharmacists needs to be strengthened.

Variation in pharmacokinetics of omeprazole and its metabolites by gender and CYP2C19 genotype in Pakistani male and female subjects.

Pharmacokinetics (PK) variation of drugs in males and females may affect therapeutic effectiveness and safety. In current study the PK differences for omeprazole and its metabolites5-hydroxy-omeprazole and omeprazole-sulphone were evaluated in males and females. The current study also considered PK comparison of Pakistani subjects using the CYP2C19 genotype as variable. A single oral dose (40mg omeprazole), open-labeland, non-controlled clinical trial was arranged. Samples were quantified using reversed phase HPLC-UV method. CYP2C19 genotype of subjects was determined by tetra primer polymerization chain reaction (PCR) assay. There was a significant increase in Cmax (from 2 to 2.9µg/ml, p=0.004**), (from 6.67 to 8.74µg-hr/ml, p=0.05*) and elimination half-life (from 1.05 to 2.1 hr, p=0.0001*) of omeprazole in females compared with males. Cmax and of 5-hydroxy-omeprazole (0.0248* and 0.0001***, respectively) and omeprazole-sulphone (0.0001*** and 0.001**, respectively) was significantly higher in females than males when compared at 95% confidence interval. The Cmax and AUC of omeprazole showed a significant raise (p=0.01* and 0.04*, respectively) in Homz PMs (Homozygous Poor Metabolizers) compared with Homz EMs (Homozygous Extensive Metabolizers) and Htrz PMs (Heterozygous Poor Metabolizers) while Cmax and AUC of 5-hydroxy-omeprazolewas significantly higher (p=0.01* and 0.04*, respectively) in Homz EMs compared with Homz PMs and HtrzPMs. AUC of omeprazole was significantly higher in females while its elimination also took longer compared with males. AUC of omeprazole was significantly higher in Homz PMs indicating that CYP2C19* displayed genetically deficient metabolism in its homozygous state.

Impact of Menopause on Pharmacokinetics of Rosuvastatin Compared with Premenopausal Women.

BACKGROUND AND OBJECTIVE: Rosuvastatin is used to treat dyslipidemia and its use is quite frequent among postmenopausal women. Menopause significantly affects the pharmacokinetics of drugs, and altered drug response and therapeutic efficacy may be anticipated in postmenopausal women compared with premenopausal women. The current study is based on assessment of differences in pharmacokinetics of rosuvastatin between pre- and postmenopausal women of Asian ethnicity. METHODS: Volunteers were administered a single oral dose of rosuvastatin 40 mg in an open-label and non-controlled pharmacokinetic study. A reversed-phase HPLC method was applied for quantification of rosuvastatin in plasma samples. Student's t test was used to compare the pharmacokinetic parameters of rosuvastatin between pre- and postmenopausal women at the 95 % confidence interval. RESULTS: The C max (premenopausal = 58.2 ± 29.1, postmenopausal = 12.2 ± 3.1 ng/ml), [Formula: see text] (premenopausal = 272.6 ± 107.3 ng·h/ml, postmenopausal = 58.8 ± 16.6 ng·h/ml), and [Formula: see text] (premenopausal = 366.1 ± 169, postmenopausal = 66.4 ± 12.9 ng·h/ml) of rosuvastatin were significantly higher (p < 0.05) in premenopausal compared with postmenopausal women. The Vd/F of rosuvastatin was significantly higher (p < 0.05) in postmenopausal women compared with women, and CL/F was also significantly (p < 0.05) faster in postmenopausal women when compared at the 95 % confidence interval. CONCLUSION: Rosuvastatin plasma level was significantly higher in premenopausal compared with postmenopausal women, which raises the question whether the latter are getting due therapeutic results, as after the menopause women experience more frequent cardiovascular problems and dyslipidemia.

Pharmacokinetics of omeprazole and its metabolites in three phases of menstrual cycle.

Omeprazole (OMP) is effective in the treatment of gastric hyperacidity and is metabolized by CYP2C19 and CYP3A4. These enzymes are modulated by estrogen and progesterone which regulate the menstrual cycle. The variations in the pharmacokinetics (PK) of many drugs like amphetamine, benzodiazepines and caffeine have been reported during menstrual cycle. In present study, the PK of the omeprazole and its metabolites was investigated during various phases of the menstrual cycle. A single oral dose, open-label, non-controlled, pharmacokinetic study of omeprazole was conducted in healthy young/premenopausal females (n = 16). The PK of omeprazole, 5-hydroxy-omeprazole and omeprazole sulphone was evaluated in three phases of menstrual cycle. The blood samples were analyzed using reversed-phase HPLC coupled with UV detector and the PK data were evaluated. The activities of CYP2C19 and CYP3A4 were determined as AUC(OH-OMP)/AUC(OMP) and AUC(OMP-SUL)/AUC(OMP), respectively. Omeprazole showed significantly (p < 0.05) higher [Formula: see text] and CL/F in follicular and menstrual phases, respectively. The [Formula: see text] of 5-hydroxy omeprazole was also significantly (p < 0.05) higher in follicular phase. The metabolic ratios (MR) of 5-hydroxy omeprazole and omeprazole sulphone were lower in follicular phase compared with the luteal phase. The present study suggests that high estrogen levels of follicular phase may result in increased absorption of omeprazole. The lower MR for 5-hydroxy omeprazole and omeprazole sulphone in follicular phase as compared to luteal phase suggests that metabolism of omeprazole is low in follicular phase as compared to luteal phase, which is progesterone-dominant phase. However, the clinical significance for these findings needs to be determined.

Pharmacokinetic study of rosuvastatin in males and females.

Rosuvastatin is used to treat dyslipidemia and is metabolized by CYP2C9 that shows variable metabolic activity in males and females. Pharmacokinetics (PK) of drugs varies in males and females that may result in altered drug response and therapeutic efficacy. In current study, PK of rosuvastatin has been evaluated in males and females. A single oral dose (40 mg rosuvastatin), open-label and non-controlled PK study was arranged. A reversed phase HPLC method was applied for quantification of rosuvastatin in serum samples. PK parameters of rosuvastatin were compared in males and females by applying student t test at 95 % confidence interval. The C max, [Formula: see text]and [Formula: see text]of rosuvastatin was significantly higher (p < 0.05) in females compared with males. The Vd/F of rosuvastatin was insignificantly higher (p > 0.05) in males compared with females while CL/F was significantly (p < 0.05) faster in males when compared at 95 % confidence interval. Rosuvastatin plasma level was significantly high in females compared with males that may be a possible reason for higher incidence of cardiac myopathy and other side effects in females. The variation in PK of drugs in males and females may require dose adjustment for maximum therapeutic effectiveness and safety.

A new HPLC method for the simultaneous determination of ascorbic acid and aminothiols in human plasma and erythrocytes using electrochemical detection.

A new, simple, economical and validated high-performance liquid chromatography linked with electrochemical detector (HPLC-ECD) method has been developed and optimized for different experimental parameters to analyze the most common monothiols and disulfide (cystine, cysteine, homocysteine, methionine, reduced (GSH) and oxidized glutathione (GSSG)) and ascorbic acid present in human plasma and erythrocytes using dopamine as internal standard (IS). Complete separation of all the targets analytes and IS at 35°C on Discovery HS C18 RP column (250 mm × 4.6mm, 5 µm) was achieved using 0.05% TFA:methanol (97:3, v/v) as a mobile phase pumped at the rate of 0.6 ml min(-1) using electrochemical detector in DC mode at the detector potential of 900 mV. The limits of detection (3 S/N) and limits of quantification (10 S/N) of the studied compounds were evaluated using dilution method. The proposed method was validated according to standard guidelines and optimization of various experimental parameters and chromatographic conditions was carried out. The optimized and validated HPLC-ECD method was successfully applied for the determination of the abovementioned compounds in human plasma and erythrocytes. The method will be quite suitable for the determination of plasma and erythrocyte profile of ascorbic acid and aminothiols in oxidative stress and other basic research studies.

Simultaneous determination of rosuvastatin and atorvastatin in human serum using RP-HPLC/UV detection: method development, validation and optimization of various experimental parameters.

A novel, precise, accurate and rapid isocratic reversed-phase high performance liquid chromatographic/ultraviolet (RP-HPLC/UV) method was developed, optimized and validated for simultaneous determination of rosuvastatin and atorvastatin in human serum using naproxen sodium as an internal standard. Effect of different experimental parameters and various particulate columns on the analysis of these analytes was evaluated. The method showed adequate separation for rosuvastatin and atorvastatin and best resolution was achieved with Brownlee analytical C18 column (150×4.6 mm, 5 µm) using methanol-water (68:32, v/v; pH adjusted to 3.0 with trifluoroacetic acid) as a mobile phase at a flow rate of 1.5 ml/min and wavelength of 241 nm. The calibration curves were linear over the concentration ranges of 2.0-256 ng/ml for rosuvastatin and 3.0-384 ng/ml for atorvastatin. The lower limit of detection (LLOD) and lower limit of quantification (LLOQ) for rosuvastatin were 0.6 and 2.0 ng/ml while for atorvastatin were 1.0 and 3.0ng/ml, respectively. All the analytes were separated in less than 7.0 min. The proposed method could be applied for routine laboratory analysis of rosuvastatin and atorvastatin in human serum samples, pharmaceutical formulations, drug-drug interaction studies and pharmacokinetics studies.