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Journal of Dental Herald
Original Article
Comparitative Evaluation Of Antibacterial Efficacy Of Herbal Extracts And Mouth Washes
Against Subgingival Plaque Bacteria. An Invitro Study.

A. Kishore Kumar , BinduPriya. S , C. Sravani , K. AmruthaSai , S. Poornodaya , N. Ravindra Reddy1 Assistant Professor, Dept of Periodontics, C.K.S Theja Institute of Dental Science and Research, Tirupathi.
2 Post Graduates, Dept of Periodontics, C.K.S Theja Institute of Dental Science and Research, Tirupathi.
3 Post Graduates, Dept of Periodontics, C.K.S Theja Institute of Dental Science and Research, Tirupathi.
4 Post Graduates, Dept of Periodontics, C.K.S Theja Institute of Dental Science and Research, Tirupathi.
5 Post Graduates, Dept of Periodontics, C.K.S Theja Institute of Dental Science and Research, Tirupathi.
6 Professor and HOD, Dept of Periodontics, C.K.S Theja Institute of Dental Science and Research, Tirupathi.
Periodontitis is a more severe destructive gum disease which may progress irreversibly in breaking down supporting periodontal structures. The
principal etiological factor in periodontal disease is dental plaque. Dental plaque is seen complex of a several hundred species of bacteria living
together forming an adherent biofilm. Bacterial flora in plaque is variable according to the site such as subgingival / supragingival .subgingival
plaque is composed of predominantly gram –ve anaerobic organisms (Tannerella forsythia, Porphyromonas gingivalis and Treponimadenticola)
are considered to be the most predominant organisms in subgingival plaque. Aim of the study was to test in vitro comparison of antibacterial activity
of herbal extract of centellaasciatica with commercially available mouth rinses against subgingival plaque bacteria. Agar well diffusion was the key
process to evaluate the antimicrobial potential of aqueous extract of the plant. MIC data showed results as bark extracts of centellaasciatica
exhibited +ve significant inhibition of bacteria even at low concentrations. Commercially available mouth washes containing synthetic and semi
synthetic active agents have several disadvantages like staining on the teeth, irritation during use, high degree of alcohol content etc. to overcome
the above disadvantages naturally occurring antibacterial herbs can be used. The herbal medicines are normally considered safer than the non
herbal medicines.
Key Words
Subgingival, Supragingival, Plaque, Predominantly, Anaerobic, Porphyromonas, Gingivalis, Treponimadenticola
appropriate method of improving the condition of the inflamed Periodontal disease is recognized as a major public health tissues. One of the major problems associated with problem throughout the world and is the most common cause conventional treatment of systemic administration of of tooth loss in adults. Periodontal disease is a general term antibiotics is the distribution of drug throughout the body, used to describe several pathological conditions that affect the which is not required and it can also give rise to toxicity supporting structures /tissues of teeth . Pain, discomfort and problems. One method of minimizing the distribution of cosmetic considerations are some of the factors that therapeutic agents in the body is through the use of local drug demonstrate severity of the problems associated with dental delivery system. Many antibacterial are applied directly to the diseases and hence, it is of utmost importance to minimize and mouth for the treatment of periodontal diseases. Mouth rinses, control dental disease . Periodontal treatment aims to cure irrigating solutions and sustained release devices are some of inflamed tissue, reduce the number of pathogenic bacteria and the local delivery systems. Mouthrinsesis most frequently eliminate the diseased pockets. Mechanical therapy, recommended to patients whose mechanical oral hygiene chemotherapy and systemic administration of antibiotics are procedures are inadequate for the plaque control. The success some of the clinical methods being utilized currently. of any mouthrinse depends on its ability in inhibiting or killing Conventional therapy includes scaling – removal of the a varied number of pathogenic oral and periodontal calculus and the plaque, curettage clearing the inflamed soft microflora.Chlorhexidine, the most commonly used tissue, and root planning - removal of necrotic tissues on the compound in various mouthrinses is a proved antimicrobial root surface.Periodontal diseases are associated with bacterial agent. This is used in combination with a variety of compounds infections; therefore antibacterial treatment seems to be an under different brand names. But prolonged use of Chlorhexidine can cause several side effects including staining of teeth, gastro-intestinal problems, gingivitis, dry mouth and Address For Correspondence:
periodontal disease some of which even help in preventing it from occurring. There are a number of herbs that can help Assistant Professor, Dept of PeriodonticsC.K.S Theja Institute of Dental Science and eliminate inflammation and infection associated with periodontal diseases. There is some evidence indicating the beneficial effect of plant extract on gingival inflammation and plaque accumulation or subgingivalperiodontopathic Journal of Dental Herald (January 2014, Issue:1, Vol.:1).
Collection Of Mouth Rinses
Centellaasiatica(L.) gotu kola is an ethnomedical plant used in Three commercially available mouthrinses with different different continents by diverse ancient cultures and tribal chemical combinations were taken as M1, M2 and M3 and groups. The Chinese prescribed the leaves in curing Ciprofloxacin (CF): Standard CF antibiotic (HIMEDIA) was leucorrhoea and toxic fever while in India, this plant is used in dissolved in 1ml of autoclaved, deionised double distilled the Ayurvedic system of medicine, usually described under the water in respective concentrations used as control.
name of Mandukaparni, to treat various diseases: asthma, bronchitis, dropsy, elephantiasis, gastric catarrh, kidney Screening For Antimicrobial Activity
troubles, leprosy, leucorrhoea, skin diseases and urethritis. It is Antimicrobial effectiveness of various mouthwashes and plant found to be quite effective in treatment of mouth ulcers, extract were assessed by Agar diffusion method by cutting healing of wounds and promote connective tissue growth. wells from seeded agar and then filling with extract obtained Asiaticoide and hypaphorine are the mixture of from plant and mouthwashes. Microbial samples were reduced pentacyclictriterpene of Centellaasiatica. It is found that according tostandard methods and the microbial topical and oral applications of asiaticoside improved wound suspensionswere prepared according to tube 0/5 McFarland healing in guinea pigs. A preliminary and follow-up study by standard(equivalent turbidity 1/5×10 CFU/ml). This Sastravaha et al investigated the effect of biodegradable chips suspension was inoculated onto Wilkins charlgren agar impregnated with Centellaasiaticaand P. granatum.
medium and MullerHinton agar mediumfor estimation of MIC as per CLSI -2012 standards. Then wells with 6 mm diameter The objective of the study is to screen and compare the were punchedusing sterile cork-borer on the culture spread antibacterial activity of plant extract of Centellaasciatica and plate.40mg/ml concentration of plant extract and 40µl of each commercially available mouth rinses on periodontal mouthwash were dispensed into wells and 40µl of 40mg/ml ciprofloxacin as positive control and the negative control was (5% DMSO). Plates were then incubated at 37?C. After Materials And Methods
incubation, the antimicrobial activities of the test materials Test microorganisms:
were determined by measuring the diameter of the zones of The periodontal organisms (T.f and P.g) were isolated from inhibition in millimeters with a HI-Antibiotic zone scale (HI- deepest pockets (> 6mm) of Periodontitis patients from the Department of Periodontics, CKS Theja College, Tirupati by removing supragingival plaque with a sterile cotton pellet and Statistical Analysis
subsequently collecting the subgingival plaque sample with a Minimum Inhibitory Concentration (MIC) was expressed in sterile paper point ISO-40 (densply) left in the periodontal terms of Mean ± SD values of all three repeats of each test pocket for 20 seconds and then the paper point was commuted sample. ‘p’ value less than 0.005 and is considered to be very to laboratory for further analysis in a sterile poly vinyl screw cap vial containing 1ml of reduced transport fluid (HI-MEDIA). The test organisms were confirmed by standard microbial, biochemical and conventional PCR analysis.
The antimicrobial efficacy of plant extract and mouth rinses
with ciprofloxacin taken as control is shown in Table-1.
Plant Material
Comparison of antibacterial efficacy of various mouthrinses with plant extracts Leaves of Centellaasciatica was collected locally in June 2012 and were identified and approved by department of Botany Sri Extract Preparation
Leaves of selected plant was extensively washed under running tap water for removal of dust particles and epiphytic hosts normally found on the surface, followed by washing with sterilized distilled water. They were further air-dried on filter paper at room temperature and then powered with the help of sterilized pestle and motar under aseptic condition. Dry Discussion
powder was further extracted by following methodologies.
Gingivitis and periodontitis, as the multifactorial diseases are mainly derived by interaction between invasions of causative Aqueous Extract
bacteria and host immune response of varied degrees. Plaque 10ml air dry powder of the respective plant parts were mixed induced gingivitis is the most common form of gingivitis and is well in 100ml of sterilized distilled water and kept at room induced by accumulation of material plaque containing more temperature for 24hrs on an orbital shaker with 500 rpm. The than 300 types of bacterial species. Removal of dental plaque solution was further filtered using muslin cloth. The filtrate that forms an important part of controlling and treating was centrifuged at 5000 rpm for 15min. The supernatant thus periodontal disease, which brings about both qualitative as obtained was filtered through Whattman’s filter no.1 under well as quantitative changes in the subgingivalmicroflora. The strict aseptic conditions and the filterer was collected in a pre- simple mechanical cleaning of the mouth has proven weighed sterilized test tube. Aqueous extracts were cotton insufficient to combat plaque formation; hence other means to plugged and stored in refrigerator at 4°c until further use.
control microbial action must also be taken. Liquids are better able to reach areas where toothbrushes would likely get stuck. Journal of Dental Herald (January 2014, Issue:1, Vol.:1).
Essential oil, chlorohexidine or CPC based product, using a Drug delivery devices: Fundamentals and applications. mouthwash will be more effective then brushing alone. Daily New York: Marcel Dekker 1998; 491-515.
use of any of the antimicrobial mouthwashes can help fight 4. Chlorhexidine Official FDA information, side effects and plaque buildup in the mouth and as a result fight gingivitis. u s e s . D r u g i n f o r m a t i o n o n l i n e h t t p : / The most common side-effect of chlorhexidine is the www.drugs.com/sfx/chlorhexidine-side-effects.html formation of extrinsic yellow-brown stains on the teeth and Plants are the natural reservoir of many 5. Lockwood B. Neutraceuticals A guide for healthcare antimicrobial agents. In recent times, traditional medicine as professionals. 2nd ed. London: Pharmaceutical Press an alternative form of health care and to overcome microbial resistance has lead the researchers to investigate the 6. Willershausen B, Gruber I and Hamm G. The influence of antimicrobial activity of medicinal plants. So as an herbal ingredients on plaque index and bleeding tendency alternative natural compounds such as plant extracts are used as natural mouth rinses typically do not contain: Alcohol, 7. Kaim JM, Gultz J, Do L and Scherer W. An in vitro Sugar, Artificial colors, Artificial sweeteners (such as investigation of the antimicrobial activity of an saccharine), Stannous fluoride, a processed form of fluoride herbalmouthrinse.J. Clin. Dent. (1998) 9: 46-48 that can stain teeth.In the present study plant extract 8. Maquart FX, Bellon G, Gillery P, Wegrowski Y, Borel JP. Centellaasciatica was tested for its antimicrobial activity Stimulation of collagen synthesis in fibroblast cultures by a against periodontal pathogens by comparing three triterpene extracted from Centellaasiatica. Connect Tissue mouthwashes taking ciprofloxacin as control. 40µl all the three mouthwashes showed excellent antimicrobial activities, of 9. S a s t r a v a h a G , Yo t n u e n g n i t P, B o o n c o n g P, those M1 showed the highest zone of inhibition, taking Sangtherapitikul P. Adjunctive periodontal treatment with ciprofloxacin as control and the plant extract Centellaasciatica Centellaasiatica and Punicagranatum extracts. of 40mg/ml showed the minimal zone of inhibition of 12mm. Apreliminarystudy. J IntAcadPeriodontol 2003; 5(4):106- The active constituents of Centella asciatica have been found 10. Poosattarbejehmir A: focusing on periodontitis as a to have antimicrobial effects. As perRishikesh et al the vasculupathay : the therapeutic possibilities from the methanol extract of Centellaasiaticashowed activity against a perspective of a dentistry student. JPMBS 2011, 12: wide a variety of microorganisms.Sastravaha et al in his 5Newman Mg, 11. TakaeHh, Klokkevold Pr, CaranzzaFa: preliminary and follow-up study investigated the effect of Clinical Periodontology. St Louis, Saunders: Eslevier; biodegradable chips impregnated with Centellaasiaticaand P. granatum in periodontal pockets showing that clinical signs of 12. Bernstein d, Schiff g,purice a. In vitro virucidal inflammation are reduced.The study concluded that effectiveness of 0.12% chlorehexidinegluconate mouth Centellaasciatica plant extract has a significant antimicrobial effect against periodontal bacteria.It can be used as a possible 13. Loe H, Schiott C. The effect of mouthrinses and topical alternative for mouth rinses to overcome the side effects of application of chlorhexidine on the development of dental chemical agents.However further extensive research and plaque and gingivitis in man. Journal of Periodontal development work should be undertaken on the active components of plant extracts and its products for their better 14. RenuSolanki Some Medicinal Plants with Antimicrobial economic and therapeutic utilization.
Activity. International Journal Of Comprehensive Pharmacy 2010 ; 01:04.
15. Rishikesh, Md. MofizurRahmanPhytochemical screening The antimicrobial potency of plants is believed to be due and in vitro antimicrobial investigation of the methanolic tannins, saponins, phenolic compounds, essential oils and extract of centellaasiaticaleaves. IJPSR, 2012; 3(9): 3323- flavonoids. This in vitro study gives us natural antimicrobial agents which can help us to control periodontal infections. The effects of these extract may benefit if incorporated in toothpastes, mouth rinses and dental products to reduce Source of Support : Nill, Conflict of Interest : None declared plaque. Further studies are required to better evaluate the effect of these extract if used as mouth rinse. In vivo clinical testing is essential to conform in vitro results. More research into herbals in dentistry can give us simple, effective solutions to periodontal diseases.
1. Schwach-Abdellaoui K, Vivien-Castoni N, Gurny R. Local
delivery of antimicrobial agents for the treatment of periodontal diseases. Eur J Pharm Biopharm 2000; 50: 83-99.
2. Friedman M, Golomb G. New sustained release dosage form of chlorhexidine for dental use.J Periodontal Res 1982; 17: 323 328.
3. Steinberg D, Friedman M. Sustained release drug delivery devices for treatment of dental diseases; In: Tyle P, Ed. Journal of Dental Herald (January 2014, Issue:1, Vol.:1).

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