A health science researcher may sometimes wonder "why statistical methods are so important in research?" Simple answer is that, statistical methods are used throughout a study that includes planning, designing, collecting data, analyzing and drawing meaningful interpretation and report the findings. Hence, it is important that a researcher knows the concepts of at least basic statistical methods used at various stages of a research study. This helps the researcher in the conduct of an appropriately well-designed study leading to valid and reliable results that can be generalized to the population. A well-designed study possesses fewer biases, which intern gives precise, valid and reliable results. There are many statistical methods and tests that are used at various stages of a research. In this communication, we discuss the overall importance of statistical considerations in medical research with the main emphasis on estimating minimum sample size for different study objectives.

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The major objective in root canal treatment is to disinfect the entire root canal system. Cleaning, shaping, and use of antimicrobial medicaments are effective in reducing the bacterial load to some extent, but some bacteria do remain behind and multiply, causing reinfection. Taking into consideration the ineffectiveness, potential side-effects and safety concerns of synthetic drugs, the herbal alternatives for endodontic usage might prove to be advantageous. Over the past decade, interest in drugs derived from medicinal plants has markedly increased. Phytomedicine has been used in dentistry as anti-inflammatory, antibiotic, analgesic, sedative and also as endodontic irrigant. Herbal preparations can be derived from the root, leaves, seeds, stem, and flowers. The PubMed database search revealed that the reference list for natural medicaments featured 1480 articles and in dentistry 173 articles. A forward search was undertaken on the selected articles and author names. This review focuses on various natural drugs and products as well as their therapeutic applications when used as phytomedicine in dentistry.

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Securinega leucopyrus (Willd.) Muell. is known as Humari in India, Katupila in Sri Lanka and Spinous fluggea in English. It is a desert climatic plant used topically in paste form for healing of chronic and non-healing wounds. Application of Katupila Kalka (paste) is used commonly in the management of acute as well as chronic wounds in Sri Lanka as a folklore medicine. The aim of this study is to evaluate the role of herbal paste of Katupila in the treatment of Dushta Vrana (chronic wound). It is a single observational innovative case study. A female aged 40 years presented with a non-healing infected wound on her right buttock with a history of 2 months. On examination, there was a rounded wound having black color necrosed tissue and slough with foul smelling, measuring about 3 inch Χ 3 inch Χ 1 inch in diameter caused by pyogenic local infection. The routine laboratory investigations were within normal limit except hemoglobin and the swab culture test of the wound bed was reported infection of Staphylococcus aureus. This case study showed effective wound healing by topical application of Katupila paste and sesame oil.

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Background: Osteoarthritis (OA) is the most common joint disorder. In Ayurveda the disease Sandhigata Vata resembles with OA, which is described under Vatavyadhi. Treatment provides symptomatic relief, but the underlying pathology remains unchecked due to the absence of effective drugs. In the management of Sandhigata Vata, all the Acharyas have described the employment of Bahya Snehan, Swedana, Abhyantara Tikta Snehapana, Basti treatment and Guggulu Prayoga. Aim: To evaluate the effect of Ksheerbala Taila Anuvasana Basti in Sandhigata Vata Materials and Methods: In the present study, 30 patients of Sandhigata Vata were given Anuvasana Basti with Ksheerabala Taila. Subjective assessment of pain by visual analog scale and swelling, tenderness, crepitus and walking velocity were graded according to their severity. Results: Significant results (P < 0.05) were found in all the cardinal symptoms - Pain (Sandhiruja), Swelling (Shotha), tenderness, crepitus and walking velocity. Radiological findings showed no significant changes. Conclusion: Anuvasana Basti with Ksheerabala Taila was significant in the subjective symptoms of Sandhigata Vata.

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Background: Parikartika resembles fissure-in-ano which is very common among all ano-rectal disorders. In Ayurvedic texts, Parikartika is described as a complication of Vamana and Virechana as well as complication of Atisara. Ksharasutra was proved successful in the management of fistula-in-ano, piles, and there is a need to try its efficacy in fissure-in-ano. Aim: To evaluate the role of Ksharasutra suturing (KSS) in fissure bed in chronic fissure-in-ano. Materials and Methods: Total 100 patients of chronic fissure-in-ano were selected and randomly divided into two groups (50 in each group). In group-A, patients were undergone by KSS; while in patients of Group-B Lord's anal dilatation followed by KSS was done under spinal anesthesia. The KSS was done once and after slough out of Ksharasutra, the wound was treated for 4 weeks and assessment of the result was done on the basis of gradation adopted. Results: The pain relief on 14 ^th day in Group-A was 86% while in Group-B 100% was observed. As on 7 ^th day in Group-A, oozing was stopped in 68% patients, while in Group-B, oozing was stopped in 82% patients. On 21 ^st day, Group-B showed more healing (85%) as compared to Group-A (69%). In this study 68% of patients were cured. Conclusion: In Group-B (KSS with Lord's anal dilatation) patients were cured early as compared to patients of Group-A (KSS alone).

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Background: India is currently witnessing rising numbers of people in the middle-class who are obese. A lot of the Indian population has started relying on processed foods that contain a huge percentage of trans-fat, sugars, and other unhealthy and artificial ingredients. Obesity is considered the core of many diseases. Increased weight carries significant health risks for some cancers, diabetes, heart diseases and strokes. Junk food, alcohol and sedentary lifestyle are leading us to silent self destruction, making one in every five Indian men and women either obese or overweight. Aim: To determine the effect of Gomutra Haritaki on Sthaulya. Materials and Methods: An observational pilot study on the effect of Gomutra Haritaki, diet control and exercise in the management of Sthaulya (obesity) was conducted on 21 patients. Enrolled patients were screened on the basis of clinical findings and allocated into two groups. Trial group received Gomutra Haritaki (6 g/day in three divided doses) while control group received placebo capsules in the same dose for 8 weeks. Result: Statistically highly significant relief was found in weight reduction (P < 0.001), and body mass index (BMI) (P < 0.01) in both groups. Control group has shown better results against trial group. Conclusion: These results prove the impact of diet and exercise in the management of Sthaulya.

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Background: Concept of Saviryta Avadhi (shelf-life) of Ayurvedic dosage forms is well-defined in classics of Ayurveda. Information on this is scattered in initial classics of Ayurveda like Charaka Samhita, but focused well after 13 ^th Century AD in texts such as Vangasena Samhita, Sharangadhara Samhita and Yogaratnakara. Though the concepts have a strong background; considering the pharmaceutical development, a need is felt to re-evaluate the age old concepts by following current norms. Aim: To evaluate shelf-life of two samples of Rasayana Churna. Materials and Methods: Raw material was procured from The Pharmacy, Gujarat Ayurved University and authenticated at Pharmacognosy Laboratory. Individual drugs were powdered and blended in specified ratio to obtain Rasayana Churna (RC). RC was levigated with decoction of Amalaki, Guduchi and Gokshura to prepare Bhavita Rasayana Churna (BRC). Both the samples were subjected to accelerated stability studies by following standard guidelines. Results: Both the samples are bitter and astringent with characteristic odor. RC is creamish brown in colour, while BRC is dark blackish brown. These Organoleptic characters were unchanged till the 6 ^th month of study. On comparision, BRC is found to be more stable than RC. Conclusion: This signifies that, Bhavana not only incorporates additional therapeutic attributes, but also helpful in increasing shelf-life.

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Background: Palatability is an important factor for choice of food by an individual. Amlarasa (sour taste) is one of the main organoleptic entities in foods of present day, which always tempts the consumer to take it now and then. According to classical Ayurvedic texts, balanced intake of Amlarasa in diet helps to maintain physiological health, but its excessive intake produces some signs and symptoms such as dentine hypersensitivity, stomatitis, halitosis, heartburn, urticaria, papule and joint inflammation. Aim: To establish the relationship between excessive use of sour predominant diets and signs/symptoms produced by it. Materials and Methods: A case-control survey study was designed wherein total of 178 volunteers were interviewed personally. Subjects with particular symptoms consider as a cases while healthy volunteers as controls. To measure the excessive intake of Amlarasa, quantity and frequency of common food articles such as mango, tomato, lime, butter milk, tamarind, curd, fermented items etc., are taken into consideration. Data was arranged in to 2 Χ 2 table and odd ratio was calculated for each symptom. Results: Odds ratio for dentine hypersensitivity, stomatitis, halitosis, heartburn, urticaria, papule and joint inflammation with 95% confidence interval were found 1.95 (0.97-3.93), 2.45 (1.12-5.40), 2.76 (0.96-7.98), 2.21 (1.09-4.53), 0.86 (0.32-2.32), 2.28 (1.02-5.05) and 4.85 (1.09-10.24) respectively. Conclusion: Study reveals that Amlarasa is a risk factor for joint inflammation, dentine hypersensitivity, stomatitis, halitosis, heartburn and papules. Study supports the Ayurvedic classical claim regarding Atiyoga of Amlarasa.

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Background: The science of wound healing is advancing rapidly, particularly as a result of new therapeutic approaches. The wound healing effect of different herbal ointments have been enormous and are in wide practice these days. Aim: To evaluate the efficacy of Panchavalkala cream over wound debridement (wound infection and microbial load). Materials and Methods: Ghanasatwa (water extract) of the individual drugs of Panchavalkala was prepared and the extract formulated as herbal ointment. This was used to treat patients of infected chronic non healing wounds. The signs and symptoms of infection were graded before and during the course of treatment. Tissue biopsy to estimate the microbial load prior to and during the course of treatment was done. Results: The clinical symptoms like Slough, swelling, redness, pain, discharge, tenderness, and malodor in wounds showed statistically significant reduction following treatment. The microbial load of the wounds was also reduced significantly. Conclusion: In most of the cases, there was a progressive reduction in the microbial load with time, during the course of treatment indicating the efficacy of the formulation in reducing the microbial load and thus controlling infection, facilitating wound healing.

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Background: Cancer screening is the main weapon for early detection at a pre-invasive or premalignant stage. It has been reported that over 12 million people use some form of tobacco, which is one of the high risk factors and has hence become an alarming world-wide problem. Aim: To evaluate the effective diagnostic screening of disease in its early stage by inexpensive method and also to evaluate the effect of indigenous mouthrinse on reversal of pre-cancerous lesions. Materials and Methods: The screening for teenagers belonging to low socio-economic status was carried out. Suspected subjects were evaluated for the reversal of the lesions by use of Ayurvedic preparation as a mouthwash. From 13 to19 years working-child population of North India was selected for the study. Screening was performed by new method-visual inspection with acetic acid. The positive subjects were further investigated by pap smear and biopsy was done as a confirmatory histopathological report. In second phase, the subjects showing positive lesions were advised indigenous anti-cancer mouth rinse and its effect was evaluated after 6 month and 9 month of prescribing the rinse. Results: The total 1095 children were screened (831 boys and 264 girls). Out of total 34 teenager boys were diagnosed, as acetowhite positive lesion. All the acetowhite positive lesions were found exclusively in males. Histological findings after 9 month use of Triphala mouth rinse revealed no changes in cells in 23 (85.2%), hyperkeratinization in 2 (7.4%), hyperkeratinization and spongiosis was evident in 1 (3.7%), mild pleomorphism in 1 (3.7%) patient. Comparative evaluation from 0-9 month showed statistically highly significant test (P < 0.01). Conclusion: Use of different forms of tobacco and betel nut showed convincing relationship between developments of oral pre-cancerous lesions. Triphala was found to have great potential for reversal of these lesions.

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Background: Dyslipidemia is a disorder of lipoprotein metabolism, including lipoprotein overproduction or deficiency and it can be understood in the parlance of the closest conditions in Ayurveda, viz. Kapha Medo Margavarana (dyslipidemia), Atisthaulya (obesity) or Meda Roga and Prameha. Asanadi Ghanavati (AG) is a modified presentation of Asanadi Gana drugs referred in Ashtanga Hridaya and Gomutra Haritaki (GH) is described in Charaka Samhita under Shotha Chikitsa and Ashtanga Hridaya in Arsha Chikitsa. Aim: To evaluate and compare the clinical effect of AG and GH in Kapha Medo Margavarana. Materials and Methods: Patients with the high lipid profile were selected and randomly divided into two groups. In Group A (n = 30), patients were administered with tablet of AG 1 g (500 mg each) thrice a day for 8 weeks and in Group B (n = 30), tablet of GH in similar dose and duration. Effect of therapy was assessed by body circumference, Body Mass Index (BMI), cardinal symptoms like Anga-Gaurava, Bharavriddhi, etc., and lipid profile parameters. Result: AG decreased the serum cholesterol by 7.12%, Serum Triglyceride (S. TG) by 7.72%, Serum Low Density Lipoprotein (S. LDL) by 11.68%, Serum Very Low Density Lipoprotein (S. VLDL) by 7.73%, and had increased Serum High Density Lipoprotein (S. HDL) by 9.52%, with moderate improvement in 14.81% and mild improvement in 70.37% of patients. The GH decreased the serum cholesterol by 6.31%, S. TG by 9.61%, S. LDL by 12.55%, serum VLDL by 8.99%, and increased S. HDL by 10.52% with moderate improvement in 3.70%, and mild improvement in 74.07% patients. Conclusion: AG and GH are suggested to be used in Kleda Bahul Samprapti Janya Vyadhi and Ama Bahul Samprapti Janya Vyadhi respectively.

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Background: Antibiotics have been in use in the treatment of bovine mastitis since decades; however, their use is associated with cost issues and human health concern. Use of herbal drugs does not generally carry these disadvantages. Many plants/herbs have been evaluated in the treatment of bovine mastitis with additional property of immunomodulation in affected mammary gland. Aim: To evaluate a topical herbal drug in two breeds of cattle for its in-vivo immunomodulatory effect on cytokines production and antibacterial activity in bovine subclinical mastitis. Materials and Methods: The response to treatment was evaluated by enumerating somatic cell count (SCC), determining total bacterial load, and studying the expression of different cytokines (interleukin [IL]-6, IL-8, IL-12, granulocyte macrophage-colony stimulating factor, interferon (IFN)-γ and tumor necrosis factor [TNF]-α). Results: The pre- and post-treatment SCC in mastitic quarters statistically did not differ significantly, however, total bacterial load declined significantly from day 0 onwards in both the breeds. Highly significant differences (P < 0.01) were observed in all the cytokines on day 0, 5, and 21 postlast treatment in both the breeds. The expression level of all the cytokines showed a significant increase on day 5, while a decrease was noticed on day 21 in both the breeds of cattle. The comparison of cytokine expression profiles between crossbred and Gir cattle revealed a significant difference in expression of IL-6 and TNF-α. However, other cytokines exhibited a similar pattern of expression in both breeds, which was non-significant. Conclusion: The topical herbal drug exhibited antibacterial and immunomodulatory activities in subclinical mastitis and thus the work supports its use as alternative herbal therapy against subclinical udder infection in bovines.

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Background: Diabetes mellitus is major metabolic disorders of carbohydrate metabolism. This leads to alter the multiple organ system. Aims: To investigate the antidiabetic and antioxidant effects of the saponin rich aqueous extract of Scoparia dulcis (SRE-SD) using alloxan-induced hyperglycemic rat model. Material and Methods: The single dose of alloxan was injected for the induction of diabetes in rats. The SRE-SD and glibenclamide were administered for 15 consecutive days from the 3 ^rd day of alloxan administration. Quantity of food and water intake was measured at day 0, and 18. Further, body weight was recorded and blood samples were collected at different time intervals that is, day 0, 3, 8, 13, and 18. The oxidative biomarkers (i.e. thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH) and nitrite (NO ²⁻) levels were also estimated in the serum sample. Results: The SRE-SD showed a remarkable dose and time-dependent changes in alloxan-induced rise in the level of food consumption and water intake, serum glucose level, TBARS, NO ²⁻ and fall in the level of GSH. Further, significant attenuation was observed at 20 and 30 mg/kg of SRE-SD treated group. Conclusions: These findings demonstrate that SRE-SD has both antidiabetic and antioxidant effects on the experimental model of diabetes in rat.

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Background: Diabetes mellitus is a persistent health problem that requires innovative strategies to improve health and needs a multifactorial approach for the treatment. Saptarangyadi Ghanavati, a formulated Ayurvedic compound consists of herbs with anti-diabetic potential. Aim: To evaluate the hypoglycemic and antihyperglycemic activities of Saptarangydi Ghanavati. Materials and Methods: For hypoglycemic activity 18 Swiss albino mice were divided into three groups (6 in each). First group served as normal control, second group is the test drug and third is the standard control group. For antihyperglycemic activity 24 Swiss albino mice were divided into four groups (6 in each). First group served as water control, second negative control, third test drug and fourth as standard control group. Test drug Saptarangydi Ghanavati was suspended in water and administered to animals at the dose of 400 mg/kg. Glibenclamide was used as reference standard in both the models at the dose of 0.65 mg/kg. Results: Saptarangyadi Ghanavati showed mild reduction in Blood Sugar Level (BSL) at all the time intervals in normoglycemic mice; however, the observed decrease of BSL was found to be statistically non significant. In antihyperglycemic activity, even though the drug failed to cease the hyperglycemia in the first hour after the glucose overload, it attenuated the same in later hours in nonsignificant extent. Conclusion: The study reveals that Saptarangydi Ghanavati has moderate hypoglycemic and antihyperglycemic effect.

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Background: Pharmacodynamics, in Ayurveda has been described in terms of Rasadipanchaka. Rasa, on one side indicates the Bhautika composition of the drug and on the other side predicts the action. Different analytical techniques, pharmaceutical processes are being used in Ayurveda for the purpose of standardization of raw drugs. Aim: In this study an attempt has been made to apply chromatographic technique in determination of Kashaya (astringent) Rasa (taste). Materials and Methods: Two important Kashaya dominant drugs Kulattha (Dolichos biflorus Linn.) and Kanchanara (Bauhinia variegata Linn.), falling under Vichitra and Samana Pratyayarabdha category respectively, were subjected to physicochemical parameters and qualitative tests followed by High-Performance Thin-Layer Chromatography (HPTLC). In light of chromatographic fingerprinting; sample preparation protocol is modified to incorporate taste threshold in correlation. Column chromatography is used for first-level discrimination technique followed by HPTLC. Kashaya Rasa Dominant Zone (KsRDZ) was separated and subjected to TLC fingerprinting. The KsRDZ fraction was designated as Botanical Reference Material (BRM) in further analysis. Results: Ash value, Alcohol and water soluble extract value were more in B variegata as compared to D biflorus. Presence of tannin in both the samples was confirmed through qualitative test. The KsRDZ fraction separated at Rf 0.46 and 0.48 for Kulattha and Kanchanara respectively. Conclusion: The results showed that the planner chromatography technique seems very useful when BRM hypothesis was adjunct to method that explains the categorization according to traditional Rasa domain classification method.

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Background: Limphonia rugosa Roth. (Merr.), family-Scrophulariaceae is considered as a botanical source of classical Ayurvedic drug Bhringaraja by the traditional practitioners of Odisha and is being used for the management of various disorders. Aim: To study the antimicrobial activity of leaf of L. rugosa. Materials and Methods: Methanol extract of L. rugosa leaf (LRLM) has been studied, at various (5, 25, 50, 100, 250 μg/ml) dilutions, against medically important human pathogenic bacteria (two Gram-positive Staphylococcus aureus, Streptococcus pyogenes and two Gram-negative-Escherichia coli, Pseudomonas aeruginosa) and two fungal strains (Aspergillus niger, A. clavatus, Candida albicans) by using the agar disc diffusion method. A zone of inhibition of extract was compared with that of different standards such as ampicillin, ciprofloxacin, norfloxacin and chloramphenicol for antibacterial activity and nystatin and griseofulvin for antifungal activity. Results: The antibacterial and antifungal activities of the LRLM increased linear with the increase in concentration of extracts. When compared with standard drugs, the results revealed that, for bacterial activity S. pyogenes and S. aureus were more sensitive and in fungal activity C. albicans was more inhibited. The range of growth inhibition zone for all the sensitive bacteria was 11-20 mm and 13-19 mm for fungal strains. Conclusion: Methanolic extract of L. rugosa leaf is having antibacterial and antifungal activities.

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Background: Persistent immune suppression is reported in Head and Neck Cancers (HNC) even after treatment and a higher recurrence rate was observed in patients with poor CD3 count. Loco regional recurrences and second primary tumours are the common forms of failure in head and neck cancers. Several agents have been tried to overcome this problem without much benefit. In Ayurveda, several plant based products have been reported to have anti-tumour and immunomodulatory properties. Aim: To test the role of Varunadi Ghritha, as an immunomodulator in apparently healthy, treated and controlled HNC patients and to evaluate its effectiveness in preventing locoregional relapses and development of second primary tumours. Materials and Methods: Total 78 patients of treated head and neck cancers were randomly selected for intervention and control group. Patients in the intervention group (n = 38) received Varunadi Ghritha, 5gms twice daily for one year and followed up to two years. Patients in the control group (n = 40) were followed up at regular intervals. Immune parameters were assessed in the peripheral blood at base line and at the end of administration of the study compound. Results: In the intervention group, mean percentage increase in CD3, CD19 and CD16 positive cells were significantly higher after the administration of the study compound compared to the control group indicating an immunomodulatory effect of the study compound. A non-significant improvement in disease control was observed in patients with advanced stage of disease in the intervention group. Conclusion: Administration of Varunadi Ghritha resulted in an increase in T cell counts in patients with treated HNC.

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Background: In the present era, because of the life-style, the disorders such as hyperacidity and gastric ulcers are found very frequently. Satwa (starch) obtained from the rhizomes of two plants namely Curcuma angustifolia Roxb. and Maranta arundinacea Linn. are used in folklore practice for the treatment of above complaints under the name Tugaksheeree. Aim: To compare the anti-ulcerogenic activity of the above two drugs in pyloric ligation induced gastric ulcer in albino rats. Materials and Methods: A total of 18 Wistar strain albino rats of both sexes grouped into three groups. Group C served as pyloric ligated control group, Group I received starch of C. angustifolia suspension and Group II received starch of M. arundinacea for seven days. On 8 ^th day pylorus was ligated. After ligation the animals were deprived of food and water and sacrificed at the end of 14 h. The collected gastric contents were used for biochemical estimation and ulcer index was calculated from excised stomach. Results: Both the test drugs showed statistically significant decrease in the volume, increase in the pH, reduced the free acidity of gastric juice and decreased the peptic activity. The starch of C. angustifolia reduced a total acidity non-significantly while M. arundinacea reduced it significantly. Among the two drugs the M. arundinacea has effectively reduced the peptic activity, which is statistically significant. M. arundinacea shown statistically significant increase of total carbohydrates. Conclusion: Both the test drugs proved anti-ulcer activity and prevents the chance of gastric ulcer. Among these two M. arundinacea is more effective.

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