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Methods for the determination of limit of detection and limit of quantitation of the analytical methods
Alankar Shrivastava, Vipin B Gupta
January-March 2011, 2(1):21-25
The quality of an analytical method developed is always appraised in terms of suitability for its intended purpose, recovery, requirement for standardization, sensitivity, analyte stability, ease of analysis, skill subset required, time and cost in that order. It is highly imperative to establish through a systematic process that the analytical method under question is acceptable for its intended purpose. Limit of detection (LOD) and limit of quantification (LOQ) are two important performance characteristics in method validation. LOD and LOQ are terms used to describe the smallest concentration of an analyte that can be reliably measured by an analytical procedure. There has often been a lack of agreement within the clinical laboratory field as to the terminology best suited to describe this parameter. Likewise, there have been various methods for estimating it. The presented review provides information relating to the calculation of the limit of detection and limit of quantitation. Brief information about differences in various regulatory agencies about these parameters is also presented here.
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Semen swallowing is safe: Oral sex focus
Rajiv Saini
July-September 2010, 1(3):30-30
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Moringa : The herbal gold to combat malnutrition
Ram Chand Dhakar, Sheo Datta Maurya, Brijendra K Pooniya, Narendra Bairwa, Manisha Gupta, Sanwarmal
July-September 2011, 2(3):119-125
This document is aimed to describing the potential benefits of the Moringa tree as a nutrient. Moringa tree can be utilized in treating the malnutrition in a local and cost-effective manner. Malnutrition causes a great deal of human suffering and is associated with more than half of all deaths of children worldwide. Malnutrition severely affects the socio-economic development of a nation because a work force that is stunted both mentally and physically may have a reduced work capacity. Thus nutrition plays an important role in the reproduction of poverty from one generation to the next. Not only is the Moringa oleifera tree extraordinary in that all parts of the tree are edible, but the most amazing aspect of the tree is its exceptionally high nutritional value. The leaves of the Moringa tree are an excellent source of vitamin A, vitamin B vitamin C and other minerals. The leaves are also an outstanding source of calcium, protein, potassium and iron. The content of amino acids such as methionine and cystine is also high. Carbohydrates, fats and phosphorous content are low making this one of the finest plant foods to be found. Present review focuses on the potential benefits of Moringa oleifera in treatment of malnutrition.
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Career opportunities in pharmacy profession: An informative note
Sandhya Bawa, Rajiv K Tonk, Jyoti A Tonk, Suresh Kumar, Sumit Singla
April-June 2010, 1(2):7-11
Pharmacy profession is always a choice of preference for the medical aspirants. There are many other medical or paramedical courses but from career opportunities point of view, the profession of pharmacy provides many job opportunities in this field. Now a day's pharma industry and profession are experiencing rapid changes due to advanced technological innovations and ideas sweeping across the world. Hence these changes give better oppor­tunities to the pharmacy profession and it is imperative to evolve a Pharmacy academic programme that suits the basic thirst of the nation like providing medicines at affordable prices, community health care etc.
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Multitude potential of wheatgrass juice (Green Blood): An overview
Swati Padalia, Sushma Drabu, Indira Raheja, Alka Gupta, Mamta Dhamija
April-June 2010, 1(2):23-28
Wheatgrass is the young grass of the common wheat plant Triticum aestivum Linn., family Poeaceae (Graminae). It is commonly known as the "green blood" due to its high chlorophyll content which accounts for 70% of its chemical constituents. It contains a plethora of vitamins, minerals, amino acids and vital enzymes like superoxide dismutase and cytochrome oxidase. The vitamin content makes it an important adjuvant in anti-allergic and anti-asthmatic treatment, while the enzymes play a pivotal role in the anticancer approach of this herbal drug. A notable feature of the wheatgrass juice is its bioflavonoid content which are the naturally occurring antioxidants and ac­count for many of its clinical utilities such as management of inflammatory bowel disease and as a general detoxi-fier. However, the most remarkable feature of the wheatgrass juice is its high chlorophyll content. Chlorophyll bears structural similarity to hemoglobin and has been found to regenerate or act as a substitute of hemoglobin in hemoglobin deficiency conditions. This might be the reason behind the utility of wheatgrass in clinical conditions like thalassemia and hemolytic anemia. The present article focuses onto the various studies emphasizing the multi­tude potentials of wheatgrass.
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Quercetin- A Flavanoid
Aarti Sharma, Himanshu Gupta
January-March 2010, 1(1):10-15
Quercetin is the most abundant form of the flavonoids. It gain attention when quercetin was found to cause DNA mutations which can then contribute to cancer treatment. Quercitrin is present in the bark of Quercus tinctoria (American Oak). It is generally available in natural sources. In this article we have tried to simplify the basic un­derstanding of quercetin, its synthesis, structure activity relationship, chemical reaction etc. It will help students to understand basic concept and chemistry of quercetin.
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Internet pharmacy: Need to be implemented in India
Ankit Anand, Neha Sethi, Gopal Sharon, George Mathew, Rajendra Songara, Prashant Kumar
January-March 2010, 1(1):16-25
In this era of science and technology, computer plays an important role in community. Today, computer is so ubiq­uitous in pharmaceutical research and development. The advent of the internet has had a significant impact on the formation of an information-driven, rapid-paced society. The number of internet users reached 150 million in only five years compared to 13 years for television and 38 years for radio. Consumer expectation for access, conven­ience, and speed has made the cyberspace superhighway a medium for knowledge exchange and for e-commerce. The internet offers a wide variety of health services and products to healthcare professionals as well as to the pub­lic. Online pharmaceutical sales have reached more than nearly $50 billion. This is a dramatic increase when com­pared to the $1.9 billion in 1999. At the click of the mouse, medications can be ordered and delivered conveniently to your door. Internet has evolved into a self-organizing media, capable of multiple interactions within. A large number of consumer products including drugs are being advertised and sold over the Internet. Though the market­ing of drugs over the Internet is an inevitable outcome of the booming e-economy, it poses unique ethical, legal and quality challenges- the prime cause being the anarchic structure of the Internet. These challenges are important from the consumer, physician and regulator perspectives. This paper begins with a summary of historical considera­tions and the shifting organization of internet pharmacy. The advantages and disadvantages of internet pharmacy practice are listed. Internet pharmacy is not only affordable but also can be source of easy availability of medicine.
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Nanoemulsion: A new concept of delivery system
Nitin Sharma, Mayank Bansal, Sharad Visht, PK Sharma, GT Kulkarni
April-June 2010, 1(2):2-6
Nanoemulsion has been identified as a promising delivery system for various drugs including biopharmaceuticals. Nanoemulsion is a heterogeneous system composed of one immiscible liquid dispersed as droplets within another liquid. The droplets size of nano emulsion is between 20 to 500 nm. Diameter and surface properties of droplets of nanoemulsion plays an important role in the biological behavior of the formulation. Small droplet sizes lead to transparent emulsions so that product appearance is not altered by the addition of an oil phase. In this paper various aspects of nanoemulsion have been discussed including advantages, disadvantages and methods of preparation. Furthermore new approaches of stability of formulation, effect of types and concentration of surfactant, process variables and method are also discussed to improve the stability of nanoemulsion formulation
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Method development and validation: Skills and tricks
Ashok Kumar, Lalit Kishore, Navpreet Kaur, Anroop Nair
January-March 2012, 3(1):3-11
Because of the introduction of a lot of drugs in the market every year, it becomes necessary to develop newer analytical methods for such drugs. Method development can take a number of forms. At one extreme, it involves adapting an existing method or making minor changes so as to make it suitable for the new application, as for developing a method for the estimation of drugs using a complex analytical technique like HPLC. After the development, there is a need of method validation. Method validation is defined as the process of proving that an analytical technique is acceptable for the intended use and this is an important requirement for analytical purpose. Validation is done according to the guidelines of ICH and FDA. Here, in this review, we have discussed method development and the various parameters used for method validation, namely accuracy, precision, limit of detection, limit of quantification, specificity, robustness, ruggedness, and range.
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An account of phytochemicals from Plumbago zeylanica (Family: Plumbaginaceae): A natural gift to human being #
Navneet Kishore, Bhuwan B Mishra, Vinod K Tiwari, Vyasji Tripathi
July-September 2012, 3(3):178-198
The semi-climbing sub-shrub Plumbago zeylanica (family: Plumbaginaceae) is a widely accepted ethnomedicine around the world including India, Pakistan, Bangladesh, Sri Lanka, and Australia. The plant is credited with potential therapeutic properties including antiatherogenic, cardiotonic, hepatoprotective, and neuroprotective properties. The present review highlights the various medicinal and pharmacological aspects along with recent updates on phytochemical contents of the plant.
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Promising future of probiotics for human health: Current scenario
Gaurav Tiwari, Ruchi Tiwari, Saurabh Pandey, Preeti Pandey
January-March 2012, 3(1):17-28
Probiotics are nonpathogenic microorganisms mostly of human origin which, when administered in adequate amounts, confer a health benefit on the host and enable to prevent or improve some diseases. Probiotics may be a natural temporary constituent of the resident intestinal microflora, but their concentration is not sufficient for therapeutic purposes. The microbiota, the intestinal epithelium, and the mucosal immune system constitute the gastrointestinal ecosystem. All three components are essential for complete functional and developmental maturity of the system. Probiotics are defined as live microbial food ingredients that have a beneficial effect on human health. The use of antibiotics, immunosuppressive therapy, and irradiation, among other means of treatment, may cause alterations in the composition and have an effect on the gastrointestinal tract flora. Therefore, the introduction of beneficial bacterial species to GI tract may be a very attractive option to re-establish the microbial equilibrium and prevent disease. The efficacy of probiotics in acute enteric infections and post-antibiotic syndromes is now established and there is emerging evidence for a role in necrotizing enterocolitis, irritable bowel syndrome, periodontal diseases, and some forms of inflammatory bowel disease.
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An assessment of groundwater quality using water quality index in Chennai, Tamil Nadu, India
I Nanda Balan, M Shivakumar, PD Madan Kumar
April-June 2012, 3(2):146-150
Context : Water, the elixir of life, is a prime natural resource. Due to rapid urbanization in India, the availability and quality of groundwater have been affected. According to the Central Groundwater Board, 80% of Chennai's groundwater has been depleted and any further exploration could lead to salt water ingression. Hence, this study was done to assess the groundwater quality in Chennai city. Aim : To assess the groundwater quality using water quality index in Chennai city. Materials and Methods: Chennai city was divided into three zones based on the legislative constituency and from these three zones three locations were randomly selected and nine groundwater samples were collected and analyzed for physiochemical properties. Results: With the exception of few parameters, most of the water quality assessment parameters showed parameters within the accepted standard values of Bureau of Indian Standards (BIS). Except for pH in a single location of zone 1, none of the parameters exceeded the permissible values for water quality assessment as prescribed by the BIS. Conclusion: This study demonstrated that in general the groundwater quality status of Chennai city ranged from excellent to good and the groundwater is fit for human consumption based on all the nine parameters of water quality index and fluoride content.
  11,673 1,268 7
Alstonia scholaris: It's Phytochemistry and pharmacology
Pawan Kaushik, Dhirender Kaushik, Neha Sharma, AC Rana
April-June 2011, 2(2):71-78
Complementary therapies based on herbal medicines are the world's oldest form of medicine and recent reports suggest that such therapies still enjoy vast popularity, especially in developing countries where most of the population does not have easy access to modern medicine. Alstonia scholaris (L.) R.Br (Apocynaceae) is an evergreen tropical tree native to Indian sub-continent and South East Asia, having grayish rough bark and milky sap rich in poisonous alkaloid. It is reported to contain various iridoids, alkaloids, coumarins, flavonoids, leucoanthocyanins, reducing sugars, simple phenolics, steroids, saponins and tannins. It has been reported to possess antimicrobial, antiamoebic, antidiarrheal, antiplasmodial, hepatoprotective, immunomodulatory, anticancer, antiasthmatic, free radical scavenging, antioxidant, analgesic, anti-inflammatory, antiulcer, antifertility and wound healing activities. In other parts of the world, it is used as a source cure against bacterial infection, malarial fever, toothache, rheumatism, snakebite, dysentery, bowl disorder, etc. Reports on the pharmacological activities of many isolated constituents from A. scholaris (L.) R.Br are lacking, which warrants further pharmacological studies.
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Multifunctional role of green blood therapy to cure for many diseases
Vipin Kumar Singhal, Anil Kumar Singhal, K Jagatheesh, K Padmavathi, N Elangoran, Om Prakash Bangr, Riya Sil
January-March 2012, 3(1):12-16
Juice of wheat (Triticum aestivum L., Poaceae) grass is termed as green blood. Wheatgrass is a variety of grass that is used like a herbal medicine for its therapeutic and nutritional properties. The aim of this study is to concise the health benefits of green blood therapy. As wheatgrass juice (WGJ) bears a close resemblance to the "hemoglobin" in our blood, the juice is called as "green blood" and the therapy using it is called as "green blood therapy". The WGJ is a complete food and contains carbohydrates, proteins, all essential minerals, and vitamins. Wheatgrass therapy is recommended for patients suffering from chronic diseases such as asthma, atherosclerosis, Parkinson's disease, joint pains, TB, constipation, hypertension, diabetes, bronchitis, insomnia, eczema, sterility, hemorrhage, obesity, and flatulence. It is also useful in the treatment of cancer. The WGJ is immediately absorbed into the bloodstream and gives energy. On an empty stomach, it is assimilated into blood in about 20 min. The energy lasts throughout the day. It is practically a fountain of youth and found to have numerous health benefits. This review is concluded that the WGJ has higher degree of curative index and this can be last alternative therapy when the all therapy will fail.
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Supercritical fluid chromatography-A Hybrid of GC and LC
Neha Sethi, Ankit Anand, Garima Jain, Kona S Srinivas, Kaushal K Chandrul
April-June 2010, 1(2):12-22
High performance specifications and unique functionality of chromatographic techniques is a demand of pharmaceutical industry and research. This leads to the origin of Supercritical Fluid Chromatography (SFC). It is a rapidly expanding analytical technique. The main feature that differentiates SFC from other chromatographic techniques is the replacement of either the liquid or gas mobile phase with a supercritical fluid mobile phase. It is considered a hybrid of GC and LC technique. High diffusion coefficient and low viscosity of supercritical fluids is responsible for high speed analysis, high efficiency and high sensitivity. Low mobile-phase flow rate, density programming and compatability with GC and LC detectors make SFC a versatile chromatographic technique in analytical re­search and development. It has a unique characteristic of analyzing thermo labile or non-volatile substances. This review highlights the role of supercritical fluid chromatography in the separation of polymers, thermally labile pesticides, fatty acids, metal chelates and organometallic compounds, chiral and achiral molecules, identification and analysis of polar samples, explosives, drugs of abuse and application of SFC in forensic science (fingerprint­ing).
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Polyhydroxyalkanoates: A way to sustainable development of bioplastics
Roopesh Jain, Susmit Kosta, Archana Tiwari
July-September 2010, 1(3):10-15
This paper reviews role of polyhydroxyalkanoates (PHAs) for sustainable development of bioplastics. PHAs are polyesters of hydroxyalkanoates with the general structural and have been investigated by biologists, chemists, engineers and other experts over the past many years. Bio-based and biodegradable plastics can form the basis for environmentally referable and sustainable alternative to current materials based exclusively on petroleum feed stocks. The result of the efforts made by scientist sought solution in form of biopolymer obtained either from microorganism or plants source and likely to replace currently used synthetic polymers as bioplastic. Applications of PHA as bioplastics, fine chemicals, implant biomaterials, and medicines have been developed. The PHA polymers promise to extend significantly the range of biomaterials as suitable alternative of plastic.
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Cross-contamination in dentistry: A comprehensive overview
Sagar J Abichandani, Ramesh Nadiger
January-June 2013, 4(1):51-58
Introduction: Cross-contamination and cross-infection can occur by direct contact with micro-organisms, indirect contact with contaminated objects, droplet transmission, and inhalation of airborne pathogens. In dentistry, operatory surfaces can routinely become contaminated with patient saliva, blood, and other fluids during treatment. Aims and Objectives: This review is aimed to identify cross-contamination and spread of infection by various means and the appropriate preventive measures to be implemented. This review will also highlight the various aspects that are neglected in various dental schools/dental practice or any dental set up that potentiate cross-contamination ultimately affecting the dentist, dental team and the patients. Materials and Methods: A review of the dental literature concerning cross-contamination was performed. Material appearing in the literature before 1996 was reviewed as exhaustively as possible and materials after 1996 were reviewed electronically. In Medline, key words like cross-contamination, sterilization, asepsis, infection, infection control, prevention were used in various combinations to obtain a potential reference for review. A total of 2245 English Language titles were found, many were repeated due to recurring searches. The headings were shortlisted and reviewed for detailed examination. Results: A comprehensive review to evaluate the methods of preventing cross-contamination in dentistry involving various aspects and challenges encountered in a dental set up was constructed which was missing in the references of the review. Conclusions: Awareness and the necessary precautions play a pivotal role in preventing the occurrence of cross-contamination. It is the responsibility of the entire dental team to work in unison to prevent the menace of cross-contamination and spread of infection.
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Polymeric micelles as a drug carrier for tumor targeting
Neha M Dand, Pranav B Patel, Anita P Ayre, Vilasrau J Kadam
July-December 2013, 4(2):94-101
Polymeric micelle can be targeted to tumor site by passive and active mechanism. Some inherent properties of polymeric micelle such as size in nanorange, stability in plasma, longevity in vivo, and pathological characteristics of tumor make polymeric micelles to be targeted at the tumor site by passive mechanism called enhanced permeability and retention effect. Polymeric micelle formed from the amphiphilic block copolymer is suitable for encapsulation of poorly water soluble, hydrophobic anticancer drugs. Other characteristics of polymeric micelles such as separated functionality at the outer shell are useful for targeting the anticancer drug to tumor by active mechanisms. Polymeric micelles can be conjugated with many ligands such as antibodies fragments, epidermal growth factors, α2 -glycoprotein, transferrine, and folate to target micelles to cancer cells. Application of heat and ultrasound are the alternative methods to enhance drug accumulation in tumoral cells. Targeting using micelles can also be done to tumor angiogenesis which is the potentially promising target for anticancer drugs. This review summarizes about recently available information regarding targeting the anticancer drug to the tumor site using polymeric micelles.
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A modification regarding the Kuppuswamy socioeconomic scale
Rahul Sharma, Narinder K Saini
January-June 2014, 5(1):84-85
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Nanotechnology based devices and applications in medicine: An overview
Elvis A Martis, Rewa R Badve, Mukta D Degwekar
January-March 2012, 3(1):68-73
Nanotechnology has been the most explored and extensively studied area in recent times. Many devices which were earlier impossible to imagine, are being developed at a lightning speed with the application of nanotechnology. To overcome the challenges offered by the most dreaded diseases, such as cancer or any disease involving the central nervous system or other inaccessible areas of the human body, nanotechnology has been proved to be a boon in making the treatment more target specific and minimizing the toxicities. This review describes a handful of important devices and applications based on nanotechnology in medicine made in recent times. This article also describes in brief the regulatory concerns and the ethical issues pertaining to nanomedical devices.
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A review on worldwide essential software resources in pharmacy
Prashant Y Mali, Shital J Panchal
January-March 2011, 2(1):11-20
Softwares are the collection of computer programs and related data that provide the instructions telling a computer what to do and thereby help to process the data as wanted. These programs are designed to address general and special purpose applications. Softwares used in pharmaceutical sciences cover wide subject areas such as pharmacology, pharmaceutical chemistry, pharmaceutics, pharmacognosy, pharmaceutical biotechnology, etc. Softwares used in pharmacology are mainly related to minimize the efforts needed in determining pharmacokinetic principles of particular drug in individuals, pathways of drug, and consequently, its adverse reactions. Applications of softwares in pharmaceutical chemistry are to elucidate various physiological properties of drugs and to predict activity values for new compounds within certain limits. They may be enormous assistance to those trying to generate the large databases from massive efforts in drug research. However, softwares used in pharmaceutics help for predicting the dissolution rate, biopharmaceutical characterization, accurate and precise stability profile, etc. of formulated dosage form. Softwares used in pharmacognosy give information on herb activity, interactions, mechanisms of action and supporting data underlying the use of herb for health. Wide applications of software in pharmaceutical biotechnology help to increase the predictability of results, identify genes, elucidate protein structure, identify genome responsible for expression of particular characteristics, etc. Hence, in the present article, we have enlisted the subject wise different software names, websites and their features used in the milieu of pharmacy.
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Drug delivery through soft contact lenses: An introduction
Gourishanker Jha, Amit Kumar
January-March 2011, 2(1):3-6
Current ophthalmic drug delivery systems are insufficient, specifically eye drops, which allow approximately 95% of the drug contained in the drops to be lost due to absorption through the conjunctiva or through the tear drainage. The use of soft contact lenses has been proposed as a method to deliver drugs to the eye in an efficient manner. The contact lenses restrict the drug from being lost to tear drainage by releasing the drug into two tear layers on either side of the contact lens, where it ultimately diffuses into the eye. By using loaded soft contact lenses, continuous drug release for extended period is possible. This paper focuses on the different methods of drug loading used throughout a polymer hydrogel.
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Pharmacological evaluation of ameliorative effect of aqueous extract of Cucumis sativus L. fruit formulation on wound healing in Wistar rats
Mithun Vishwanath K Patil, Amit D Kandhare, Sucheta D Bhise
October-December 2011, 2(4):207-213
Aim: The aim of present investigation was to formulate and evaluate the ameliorative effect of aqueous extract of Cucumis sativus L. fruit cream formulation on experimentally induced wounds in rats. Materials and Methods: The cream was formulated using soft white paraffin base containing 2.5%, 5%, and 10% w/w of aqueous extract of of Cucumis sativus L. fruit. Excision wounds of size 300 mm 2 and 2 mm depth were used for the study of rate of contraction of wound and epithelization. All the three formulations were evaluated for various pharmaceutical parameters such as pH, viscosity, spreadability, and acute skin irritation study. Epithelialization period, wound contraction, scar width, and histopathological evaluation parameters were used for pharmacological evaluation of wound healing activity of the formulation. Statistical Analysis: All the results were expressed as mean±SEM. Data analysis was performed using GraphPad Prism 5.0 software (GraphPad, San Diego, CA, USA). Statistical comparisons were made between drug-treated groups and disease control animals. Data of disease activity index were analyzed using one-way analysis of variance; Dunnett's multiple range test was applied for post hoc analysis, whereas data of wound area and percent wound contraction were analyzed using two-way repeated analysis of variance, Bonferroni's multiple range test was applied for post hoc analysis. A value of P<0.05 was considered to be statistically significant. Results: Cream formulation of AECS when applied topically did not show any sign and symptoms of skin irritation. The treatment with aqueous extract of C. sativus fruit cream formulation (2.5%, 5%, and 10% w/w) resulted in significance decrease (P<0.05, P<0.001, and P<0.001, respectively) in wound area, epithelization period, and scar width, whereas rate of wound contraction significance increased (P<0.001 respectively) when compared with control group animals. Conclusion: The present investigation demonstrates that by virtue of its antioxidant property and presence of the flavanoids content in C. sativus may responsible for wound contraction and elevated rate of epithelization in wound healing in laboratory animals.
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Bio-inspired nanomaterials and their applications as antimicrobial agents
Smita Sachin Zinjarde
January-March 2012, 3(1):74-81
In the recent decades, the interdisciplinary field of nanotechnology has expanded extensively. A variety of nanoparticles (NPs) have been used for a number of specialized applications. In this era facing a major problem of microorganisms developing antibiotic resistance, NPs are a lucrative option. Most physical and chemical processes of NP synthesis are associated with drawbacks and bio-inspired NPs have now become popular. This review summarizes the recent developments on the biosynthesis, characterization, and applications of NPs with particular reference to their use as antimicrobial agents. Reviewed here is the synthesis of gold and silver NPs (AgNPs) by a variety of biological forms and biomolecules as well as their effectiveness toward different fungal and bacterial pathogens. The use of gold NPs (bio-inspired by plants, fungi, and bacteria) and AgNPs, synthesized by carbohydrates (of plant, animal, and microbial origin), plant parts (bark, callus, leaves, peels, and tubers), fungi, and bacteria have been highlighted. In addition, the use of zinc oxide NPs (although not bio-inspired) as novel antimicrobial agents have also been discussed.
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Phenotypic methods for detection of various β-lactamases in Gram-negative clinical isolates: Need of the hour
Neena V Nagdeo, Navinchandra M Kaore, Vilas R Thombare
October-December 2012, 3(4):292-298
Background: Many clinical laboratories have problems detecting various β-lactamases. Confusion exists about the importance of these resistance mechanisms, optimal test methods, and appropriate reporting conventions. It is more imperative to use various phenotypic methods for detection of various β-lactamases in routine microbiology laboratory on day-to-day basis to prevent antimicrobial resistance by evidence-based judicious use of antimicrobials. Aims: In view of the multidrug-resistant organisms being reported world over, we planned a cross-sectional prospective analytical study to determine resistance mechanism by various β-lactamases in Gram-negative clinical isolates using various phenotypic methods. Materials and Methods: All nonrepeat, nonenteric clinical isolates of Gram-negative bacilli, resistant to at least two third-generation cephalosporins, were first screened by Novel disc placement method, and isolates showing multiple mechanisms of resistance and reduced zone of inhibition for imipenem were further confirmed for AmpC and metallo β-lactamases. Statistical Analysis: All the data was managed and analyzed in Microsoft Excel. Results: Out of 807 isolates tested, as many as 795 (98.51%) revealed the presence of extended-spectrum β-lactamases (ESBLs). Only 10 isolates of Escherichia coli and 2 of Klebsiella pneumoniae did not show production of ESBL. A total of 450 (55.76%) isolates produced single enzyme,while 345 (42.75%) strains revealed multiple enzyme production simultaneously. Only ESBL production was seen in 315 (39.03%) strains, only AmpC in 75 (9.29%) and only MBL in 60 (7.44%) strains, while ESBL and AmpC together were seen in 219 (27.14%) and AmpC plus MBL in 92 (11.40%) strains. However, ESBL plus MBL were never observed together. All three enzymes were simultaneously detected in 34 (4.21%) strains. Conclusion: This innovative method of disc placement makes it easy, affordable, and reliable method for routine use by basic microbiology laboratories for detection of various β-lactamases, pending confirmation for AmpC and metallo β-lactamase by three-dimensional test and double disc potentiation test, respectively.
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