Assessment of antidiabetic potential and phytochemical profiling of Rhazya stricta root extracts.

BACKGROUND: Diabetes mellitus is a chronic disease characterized by hyperglycemia that may occur due to genetic, environmental or lifestyle factors. Natural remedies have been used to treat diabetes since long and many antidiabetic compounds of varied efficacies have been isolated from medicinal plants. Rhazya stricta has been used for decades for the treatment of diabetes mellitus and associated ailments. Considering the folkloric use of R. stricta against diabetes, it was aimed to investigate the effectiveness of its root extracts against diabetes through in vitro assays and in vivo studies using animal model along with phytochemical profiling through GCMS. METHODS: Various fractions of Rhazya stricta obtained through column chromatography were evaluated for a variety of assays including α-glucosidase, Dipeptidyl peptidase-IV (DPP-IV), ß-secretase and Glucagon-like peptide-1 (GLP-1) secretion studies. For the in vivo studies the alloxan-induced diabetic mice were treated with root extracts and blood glucose levels, HbA1C, and other biochemical markers along with the histological study of the liver were done. The phytochemical identification was performed using an Agilent 7890B GC coupled to a 7010 Triple Quadrupole (MS/MS) system. GraphPad Prism software version 5.01 was used for statistical analysis. RESULTS: Majority of the extract fractions showed excellent results against diabetes by inhibiting enzymes DPP-IV (Up to 61%) and ß-secretase (Up to 83%) with IC50s 979 µg/ml and 169 µg/ml respectively with increase in the GLP1 secretion. The results of in vivo studies indicated a marked reduction in blood glucose and HbA1c levels along with positive effects on other parameters like lipid profile, liver functions and renal functions of extract-treated mice as compared to control. The histological examination of the liver demonstrated hepatoprotective effects against diabetes led changes and various classes of phytochemicals were also identified through GCMS in different fractions. CONCLUSION: The results revealed strong antidiabetic activity of R. stricta root with the potential to protect body organs against diabetic changes. Moreover, a variety of phytochemicals has also been identified through GCMS that might be responsible for the antidiabetic potential of Rhazya stricta root.

Pharmacological evaluation of Rhazya stricta root extract / Evaluación farmacológica del extracto de raíz de Rhazya stricta

The present study aimed to screen the Rhazya stricta Decne root for its antihyperglycemic and antioxidants potential through invitro assays along with phytochemical and elemental analyses. The crude extract was prepared through maceration and fractionated using solvent-solvent extraction technique. The spectroscopic studies indicated the presence of various phytochemical classes in the extract and its fractions. The antioxidant assays showed notable results along with a good concentration of phenolic and flavonoid contents. Enzyme inhibition assays demonstrated glucose-lowering effects by inhibiting the enzyme activity which could reduce post-prandial blood glucose level. The Dipeptidyl peptidase-IV (DPP-IV) inhibition assay results showed the novel DPP-IV inhibition activity of the plant extract and all fractions showed noteworthy enzyme inhibition and antihyperglycemic activity. Conclusively, the Rhazya stricta root extract displayed its antioxidant and antihyperglycemic potential due to the presence of various classes of phytochemicals and micro-nutrients.

El presente estudio tuvo como objetivo examinar la raíz de Rhazya stricta Decne por su potencial antihiperglicémico y antioxidante a través de ensayos in vitro junto con análisis fitoquímicos y elementales. El extracto crudo se preparó por maceración y se fraccionó usando una técnica de extracción solvente-solvente. Los estudios espectroscópicos indicaron la presencia de varias clases fitoquímicas en el extracto y sus fracciones. Los ensayos antioxidantes mostraron resultados notables junto con una importante concentración de contenido fenólico y flavonoide. Los ensayos de inhibición enzimática demostraron efectos reductores de la glucosa al inhibir la actividad enzimática que podría reducir el nivel de glucosa posprandial en sangre. Los resultados del ensayo de inhibición de Dipeptidyl peptidase-IV (DPP-IV) mostraron la nueva actividad de inhibición de DPP-IV del extracto de la planta y todas las fracciones mostraron una notable inhibición enzimática y actividad antihiperglicémica. En conclusión, el extracto de raíz de Rhazya stricta Decne mostró su potencial antioxidante y antihiperglicémico debido a la presencia de varias clases de fitoquímicos y micronutrientes.

Microbial and chemical analysis of illicit drugs samples confiscated from different areas of PakistanMicrobial and chemical analysis of illicit drugs samples confiscated from different areas of Pakistan.

The microbial and chemical analysis of illicit drug samples from different areas of Pakistan i.e. Quetta, Karachi, Lahore and Islamabad was conducted in a cross-sectional study at National Institute of Health, Islamabad. The drug samples were confiscated by Anti Narcotics Force (ANF), Pakistan. Microbial analysis was done by estimating bioburden which revealed the presence of gram negative and positive bacteria's, fungus, Streptococcus, Staphylococcus species. Trypton soya agar was used for total aerobic count, MacConkey agar for gram-negative bacteria, Sabouraud dextrose agar for fungus and Vogel-Johnson agar for Streptococcus and Staphylococcus species. Colour tests were applied to identify the drug samples. Qualitative and quantitative analysis of suspected samples of Heroin, morphine, cocaine and acetic anhydride was made by employing different chromatographic techniques i.e. Thin-layer chromatography (TLC) and High-performance liquid chromatography (HPLC). The samples were found to be adulterated with paracetamol, diazepam and Dextromethorphen. Acetic anhydride was adulterated with hydrochloric acid (HCl). There is lack of information providing structured advice on responses to the consequences of illicit drug adulteration. Robust and rehearsed interventions and communication strategies would provide a basis for response for a wide variety of organisations. Research into the usefulness of media warnings about adulteration of illicit drugs is required.

EVALUATION OF DRUG USE INDICATORS FOR NON-COMMUNICABLE DISEASES IN PAKISTAN.

Irrational drug use practices are a burden to healthcare facilities. Poor prescribing practices affect the overall management and cost of treatment of non-communicable diseases that are the major cause of mortality and morbidity worldwide. In an effort to improve prescribing practices, this study was designed to assess prescribing, consultation and facility indicators in healthcare facilities of Punjab and Sindh provinces of Pakistan from December 2012 to December 2013. In this cross-sectional study, random and convenient sampling were used to collected data from both private and public healthcare facilities. Quantitative data were collected using structured questionnaire, observations and prescription analysis, whereas qualitative information on factors influencing prescribing practices was obtained by interviewing medical practitioners. A total of 13693 prescriptions were obtained from 500 patient-prescriber encounters. Results show that history taking, physical examination and diagnoses were adequate while generic prescribing was four-fold less than drugs prescribed by brands. Average number of drugs prescribed was 4.63 with more prescribing tendency in private facilities. 45.07% prescription costs were less than Rs. 150. Sulfonylureas, statins and ACE inhibitors were highly prescribed drugs for diabetes, hyperlipidemia and hypertension. Prescribing practices were dominantly influenced by severity of disease (73% Punjab; 81% Sindh), patient age (75% Punjab; 68% Sindh) and availability of drugs (62% Punjab; 56% Sindh) whereby 91% practitioners in Sindh and 52% in Punjab rely on medical representatives as the source of drug information. Moreover, the pharmacy and therapeutic committees in all facilities were non-functional along with non-availability of essential drug list in 87% health facilities. Thus, there are considerable opportunities to improve the rational use of medicines in Pakistan including low prices for generics, physician education, prescribing guidelines and formularies.

COMPARATIVE BIOAVAILABILITY ANALYSIS OF ORAL ALENDRONATE SODIUM FORMULATIONS IN PAKISTAN.

Alendronate sodium, a bisphosphonate drug, it is used to treat osteoporosis and other bone diseases. The present study was designed to conduct comparative bioavailability analysis of oral formulations of aledronate sodium through an open-label, randomized, 2-sequence, 2-period crossover study. Healthy adult male Pakistani volunteers received a single 70 mg dose of the test or reference formulation of alendronate sodium followed by a 7 day washout period. Plasma drug concentrations were determined using a validated HPLC post column fluorescence derivatization method. AUC(01,) AUC(0-8,) C(max). and T(max) were determined by non-compartmental analysis and were found within the permitted range of 80% to 125% set by the US Food and Drug Administration (FDA). Results show that both in vitio and in vivo assays of all test brands were within the spec- ification of the US Pharmacopoeial limits and were statistically bioequivalent. No adverse events were reported in this study.

Antioxidant activity and inhibitory effect of some commonly used medicinal plants against lipid per-oxidation in mice brain.

BACKGROUND: The present study compares the protective properties of aqueous extracts of six medicinal plants, Phyllanthus emblica, Terminalia chebula (black and yellow), Terminalia arjuna, Balsamodendron Mukul and Alium sativum against lipid per-oxidation in mice brain. METHODS: The antioxidant activities were analyzed by lipid per-oxidation assay, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical assay, total antioxidant activity and metal chelation. RESULTS: The extracts (fruits and bark) showed inhibition against thiobarbituric acid reactive species (TBARS) induced by pro-oxidant (10 µM FeSO4) in mice brain. Moreover, the free radical scavenging activities of the extracts was evaluated by the scavenging of DPPH radical (IC50, 23.23 ± 1.2 µg/ml (Phyllanthus emblica), 20.24 ± 0.9 µg/ml (Terminalia chebula yellow) and 17.33 ± 1.1 µg/ml (Terminalia chebula black), 19.44 ± 0.45 µg/ml (Terminalia arjuna), 56.59 ± 2.1 µg/ml (Balsamodendron Mukul) and < 200 µg/ml (Alium sativum). CONCLUSION: The higher antioxidant and inhibitory effect of Terminalia chebula black in this study could be attributed to its significantly higher phenolic content, Fe(II) chelating ability, reducing ability and free radical scavenging activity. Therefore oxidative stress in brain could be potentially prevented by the intake of these plants.

Comparative phytochemical, hepatoprotective and antioxidant activities of various samples of Swertia Chirayita collected from various cities of Pakistan.

Medicinal plants are crucial for about 80% of the world population in developing and developed countries for their primary and basic health care needs owing to better tolerability, superior compatibility with human body and having lesser side effects. The present study was conducted on various solvent extracts of three plant samples of Indian and Nepali origin Swertia Chirayita (Roxb.) Buch-ham (Chiratia) collected from various places to establish their comparative phytochemical analysis, chromatographic profile, hepatoprotective and antioxidant activities. Nepali Swertia Chirayita was found to have finest Chromatographic profile (TLC). Phytochemical analysis revealed Alkaloids, flavonoids, saponins, ascorbic acid, glycosides, steroids and triterpenoids in all samples. Different solvent fractions of the methanolic plant extracts of Swertia chirayita were assessed for hepatoprotective activity by carbon tetrachloride-induced liver damage in rats. The grade of protection was measured by using biochemical parameters such as serum glutamate oxalate transaminase (SGOT/AST), alkaline phosphatase (ALP), serum glutamate pyruvate transaminase (SGPT/ALT) and total bilirubin. The in-vitro antioxidant activity of the extracts was also evaluated by the 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay. The methanolic and aqueous extracts, at a dose of 200mg/kg and 300mg/kg, produced significant (p<0.05) hepatoprotection by decreasing the activities of the serum enzymes and bilirubin while there were marked scavenging of the DPPH free radicals by the fractions. Decreased observed in the biochemical parameters suggests that the plant extracts possesses hepatoprotective as well as antioxidant activities without any significant variation amongst them. These activities reside mainly in the methanolic extract of whole plant.

Review: an exposition of medicinal preponderance of Moringa oleifera (Lank.).

Medicinal plants are believed to be a precious natural reservoir as they are assumed to have paranormal effects for the mankind. Moringa oleifera grows throughout most of the tropics and has numerous industrial and medicinal uses. This review acquaints with the consequence of fera (Moringaceae), a fast growing medicinal plant wide spread in tropical regions with height ranging from 5-10m. It has an enormous nutritional worth due to existence of vitamins and proteins. It is subsisted with many constituents. Its oil consists of oleic, tocopherols, stearic, palmitic, behenic and arachidic acid. Flavanoids and phenolics such as gallic acid, chlorogenic acid, ferulic acid, kaempferol, ellagic acid, quercetin and vanillin are present by means of leaf extract, being richest in phenolics and subsequent fruit and seed extract respectively, that are accountable for antioxidant activity of plant. Seeds have been pragmatic with active components as novel O-ethyl-4- (α -L-rhamnosyloxy) benzyl carbamate together with seven known compounds, 4 (α -L-rhamnosyloxy)-benzyl isothiocyanate, niazimicin, niazirin, ß-sitosterol, glycerol-1- (9 -octadecanoate), 3 -O- 6 -O-oleoyl- ß -D-glucopyranosyl-b-sitosterol, and ß - sitosterol- 3-X-O -ß -D-glucopyranoside , that have been discerned to inhibit EBV-EA (Epstein- Barr virus-early antigen), that is persuaded by the cancer promoter. M. oleifera leaves, gums, roots, flowers as well as kernels have been unanimously utilized for managing tissue tenderness, cardiovascular and liver maladies, normalize blood glucose and cholesterol. It has also profound antimicrobial, hypoglycemic and anti-tubercular activities.

Critique of medicinal conspicuousness of Parsley(Petroselinum crispum): a culinary herb of Mediterranean region.

WHO estimates, around 80% of the especially developing world is indigent on complementary and alternative medicines which are prodigiously derived from herbal material. Parsley (Petroselinum crispum) is an important culinary herb originated from the Mediterranean region. It possesses small and dark seeds with volatile oil content. Petroselinum crispum is now planted throughout the world due to its usage in food industry, perfume manufacturing, soaps, and creams. Its main constituents subsume coumarins, furanocoumarins (bergapten, imperatori), ascorbic acid, carotenoids, flavonoids, apiole, various terpenoic compounds, phenyl propanoids, phathalides, and tocopherol. Due to these constituents, it has been annunciated to possess a number of possible medicinal emblematics including, antimicrobial, antianemic, menorrhagic, anticoagulant, antihyperlipidemic, antihepatotoxic, antihypertensive, diuretic effects, hypoglycaemic, hypouricemic, anti oxidative and estrogenic activities. In Morocco, Parsley is mostly used as an elixir to treat arterial hypertension, diabetes, cardiac and renal diseases. Antioxidant and antibacterial activities of parsley, made it propitious in food systems. Its ELI17 gene has been corroborated as a particularly fast-responding gene. There is a requisite for extensive research to avail the maximal benefits of this significant medicinal plant. The aim of this review paper is to divulge the chemical constituents of parsley that are explicitly related to substantial medicinal facets.

Accentuating the prodigious significance of Eclipta alba - an inestimable medicinal plant.

Eclipta alba is a small branched perennial herb, which has been used as a traditional medicine in different countries mainly in tropical and subtropical regions of the world. The plant E. alba plays a significant role in the ayurvedic, traditional and unani systems of medicine. It is popularly known as "Bhringaraj". The herb has been known for its medicinal value and has been used as an analgesic, antimytotoxic, antihepatotoxic, antibacterial, antioxidant, antihaemorrhagic, antihyperglycemic and immunomodulatory and also recognized as a reincarnated plant. Broad range of chemical constituents have been detached from E. alba including coumestans, alkaloids, thiopenes, flavonoids, polyacetylenes, triterpenes and their glycosides. Pharmacological activities have been seen in the metabolites and extracts of this plant. Therefore this herb produces robust curative lead compounds, which would be propitious for humanity. The purpose of this review recapitulates all data related to E. alba considering its prodigious medicinal importance.

Real time and accelerated stability studies of Tetanus toxoid manufactured in public sector facilities of Pakistan.

Tetanus is an acute illness represented by comprehensive increased inflexibility and spastic spasms of skeletal muscles. The poor quality tetanus toxoid vaccine can raise the prevalence of neonatal tetanus. WHO has taken numerous steps to assist national regulatory authorities and vaccine manufacturers to ensure its quality and efficacy. It has formulated international principles for stability evaluation of each vaccine, which are available in the form of recommendations and guidelines. The aim of present study was to ensure the stability of tetanus vaccines produced by National Institute of Health, Islamabad, Pakistan by employing standardized methods to ensure constancy of tetanus toxoid at elevated temperature, if during storage/transportation cold chain may not be maintained in hot weather. A total of three batches filled during full-scale production were tested. All Stability studies determination were performed on final products stored at 2-8°C and elevated temperatures in conformance with the ICH Guideline of Stability Testing of Biological Products. These studies gave comparison between real time shelf-life stability and accelerated stability studies. The findings indicate long-term thermo stability and prove that this tetanus vaccine can remain efficient under setting of routine use when suggested measures for storage and handling are followed in true spirit.

Appraisal of multifarious brands of Cephradine for their in vitro antibacterial activity against varied microorganisms.

The astounding and exceptional growth of generic pharmaceutical Industry in Pakistan has raised certain questions for drug regulatory authorities contemplating their efficacy and quality. The current study focuses on assessing the in-vitro antimicrobial activity of 24 brands of Cephradine 500mg capsules against 4 different strains by employing standardized methods. Disk diffusion method was performed on all brands to look into the susceptibility and resistance patterns. Standard disk of 5µg Cephradine powder were used during evaluation. The zones of inhibitions were ranged from 24-40mm against S. aureus, 24-40mm against E. coli, 20-25mm against K. pneumonia and 19-23mm P. mirabilis. On the basis of mean value, the multinational brands were found to have better zone of inhibitions and were better than local Pharmaceutical companies but ANOVA cooperative study showed that all brands of Cephradine showed similar comparable results. Further investigations by employing MIC method, quality of raw material with special emphasis on the shelf-life, excepients and method of manufacturing will be needed to obtain more authenticated results. The price of National and Multinational brands ranges from Rs.156.00-212.00 for 10 capsules. It is concluded that Public health is at risk because of noticeable growing widespread curse of the manufacture and trade of sub-standard or below par pharmaceuticals. The pecuniary accountability of management of pharmaceutical agents is additionally apparent. The results of the study need to be made public to boost the confidence of medical profession about the quality of locally manufactured pharmaceuticals. It will succour the foreign exchange being incurred on the trade in of medicines.

Enzymatic saccharification and lactic acid production from banana pseudo-stem through optimized pretreatment at lowest catalyst concentration.

This work estimates the potential of banana pseudo-stem with high cellulosic content 42.2-63 %, for the production of fermentable sugars for lactic acid production through statistically optimized pretreatment method. To evaluate the catalyzed pretreatment efficiency of banana pseudo stem based on the enzymatic digestibility, Response Surface Methodology (RSM) was employed for the optimization of pretreatment temperature and time using lowest concentrations of H2SO4, NaOH, NaOH catalyzed Na2S and Na2SO3 that seemed to be significant variables with P<0.05. High F and R (2) values and low p-value for hydrolysis yield indicated the model predictability. The optimized condition for NaOH was determined to be conc. 1 %, temperature 130 (o)C for 2.6 hr; Na2S; conc. 1 %, temperature 130 (o)C for 2.29 hr; Na2SO3; conc. 1 %, temperature 130 (o)C for 2.41 hr and H2SO4; conc. 1 %, temperature 129.45 (o)C for 2.18 hr, produced 84.91 %, 85.23 %, 81.2 % and 76.02 % hydrolysis yield, respectively. Sulphuric acid provided 33+1 gL(-1) reducing sugars in pretreatment step along with 38+0.5 gL(-1) during enzymatic hydrolysis. Separate hydrolysis and fermentation of resulting sugars showed that the conversion of glucans into lactic acid reached 92 % of the theoretical yield of glucose.

Pakistani pharmacy students' perception about complementary and alternative medicine.

OBJECTIVES: To assess Pakistani pharmacy students' perceptions of complementary and alternative medicine (CAM), the frequency with which they use CAM, and barriers to use of CAM. METHOD: A CAM health belief questionnaire was administered to 595 students enrolled in a 5-year doctor of pharmacy program (PharmD) in Pakistan. RESULTS: Attitudes of students towards CAM were positive. Lack of evidence supporting CAM practices was considered to be the major barrier toward more students using CAM. A majority of students (79%) agreed that clinical care should integrate conventional medicine and CAM practices. Many CAM-based therapies, such as dietary supplements, massage, herbal medicines, and homoeopathic medicines were used by the students. Significant gender differences in attitude were observed, with male students having more conservative attitudes toward CAM use. A high percentage of students desired more training in CAM. CONCLUSIONS: Pakistani students exhibited positive attitudes about the value of CAM and most felt that CAM should be included in the PharmD curriculum.

Preparation and characterization of chloramphenicol niosomes and comparison with chloramphenicol eye drops (0.5%w/v) in experimental conjunctivitis in albino rabbits.

Niosomes has gained tremendous popularity as ultimate drug carrier. Lot of research work is being carried out on preparation of niosomes for ophthalmic use having no significant effect on vision and its sustained release pattern. Chloramphenicol niosomes were prepared using two different ratios of cholesterol, drug and surfactant, termed as EIN-1, EIN-2 by ether injection method and their entrapment efficiency, particle size. The in vitro drug release pattern was observed for ten hours. The EIN-2 showed 90% entrapment and released 81% of entrapped drug after 10 hours. Zeta potential & viscosity were determined and in-vivo comparison was made with Chloramphenicol eye drops where it exhibited Cmax of 15 µ g/ml. Stability studies were done to determine shelf life. MIC of selected strain of S. aureus was also determined. EIN 2 niosomal suspension was compared with Chloramphenicol eye drops in experimental conjunctivitis in albino rabbits. In-vitro studies are encouraging as niosomes released about 75% of total entrapped drug by EIN-1 and 81% of total entrapped drug by EIN 2. In vivo study shows that niosomes released the drug in eye in acceptable range and showed a sustained release pattern without affecting the vision. Niosomes were found ultimate ophthalmic drug carriers capable to release drug in sustained and determined pattern.

Trends in the use of complementary and alternative medicine in Pakistan: a population-based survey.

OBJECTIVES: The objective of this study was to evaluate the extent of use of complementary and alternative systems of medicine by different segments of society and to identify the diseases in which they have been found to be effective. METHODS: This was a questionnaire-based descriptive study carried out from April 2002 to March 2004, in selected urban and rural areas of four provinces of Pakistan representing general strata of population from various socioeconomic conditions. RESULTS: The overall trend in Pakistan shows that 51.7% (CI 54.3-49.1) chose complementary and alternative medicine (CAM) while 48.3% (CI 50.71-48.89) chose biomedicine. Of those who chose CAM, 20% (CI 21-19) also used biomedicine as well; 16% (CI 16.8-15.2) homeopathy, 12.4% (CI 13.02-11.78) unani medicine, 2.1% (CI 2.20-1.99) mind-body medicine (faith healing), 0.9% biologically based practices (home remedies, diet and nutrition) 0.05% energy medicine (Reiki), 0.05% Traditional Chinese Medicine, and 0.02% aromatherapy. CONCLUSIONS: About half of the studied population used CAM. The population estimates of use of CAM are within the range reported elsewhere. It reflects an increasing popularity of CAM in Pakistan as well. Combined use of biomedicine with CAM was common and often patients did not reveal the use of CAM to the biomedicine practitioners.