Herbal Sources Research Articles

EVALUATION OF SOME TRADITIONAL THERAPEUTIC PROPERTIES OF USNEA LONGISSIMA (ASCOMYCOTA, LICHENIZED FUNGI): ANTIMICROBIAL, ANTIQUORUM AND ANTIOXIDANT

Usnea longissima, fruticose lichen growing in the temperate region, is well known traditional medicinal herb and a reservoir of some unique secondary metabolites. Traditionally it has been commonly used as an antimicrobial agent for the treatment of ailments like tuberculosis, ulcers and skin diseases, etc. In our study, we evaluate antimicrobial, antiquorum sensing (QS) and antioxidant activity of Usnea longissima secondary metabolites extracted in acetone, methanol, and 70% hydroalcoholic solvents. Antimicrobial activity of extracts was evaluated against fungus (Aspergillus niger, Candida albicans) and six bacterial (Agrobacterium tumefaciens, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aerogenosa, Staphylococcus aureus, Streptococcus mutans) strains. Whereas anti-QS activity was tested against biosensor strain Chromobacterium violaceum (CV) MCC 2290 and antibiofilm activity was checked in two gram-negative bacteria (P. aeruginosa and E. coli), two gram-positive bacteria (Bacillus cereus and Staphylococcus aureus) and fungus (Candida albicans). Results of experimental data indicated that methanolic extract exhibited maximum activity against A. tumefaciens (24±0.2 mm) and C. albicans (30±0.3 mm) with the minimum inhibitory concentration of 300µg/mL and 200µg/mL respectively. Interestingly, we also observed that all extracts effectively inhibited quorum sensing in C. violaceum and biofilm formation in Bacillus cereus, Escherichia coli, Pseudomonas aerogenosa, Staphylococcus aureus, at the concentration (100-300 µg/mL), for Candida albicans (50-150 µg/mL). Usnea longissima extract showed strong reducing power and hydroxyl radical activity and low DPPH scavenging activity. The present study suggested that U. longissima possesses potential antimicrobial and antiquorum sensing activity. These biological properties could be attributed to its significant antioxidant potential which validates its traditional use as an antimicrobial herbal source.

Concept of standard raw drug substitution in Traditional Siddha Medicine - A Review

Classical Siddha medicine recommends the usage of functionally parallel substitute raw drugs in the scarcity of the original material. The concept of standard raw drug substitution is designated by the Tamil term Matru sarakku, which emphasizes selecting alternatives either in herbal or non-herbal sources based on certain attributes. Functionally similar materials are suggested for balancing the deficit based on such parameters. Literature sources from Siddha medicine and other allied subjects were explored and documented for the inclusive understanding of the concept of ideal substitution. Few of the standard substitutes mentioned in the classical Siddha literature were evaluated in a scientific account for its justification. An outlook on different specimens as described in the ancient texts of Tamil medicine indicates the usage of substitutes from numerous plants, animals, and other metallo-mineral ingredients. The rationale of selecting alternatives primarily depends on the equivalent following traditional attributes like organoleptic entities, potency, division, general properties, specific actions, and medicinal uses of the material shared between the raw drugs. This must undergo a systematic evaluation by pharmacognostic and phytochemical studies to justify current practices of substitution. A systematic document in these lines will give a proper guideline for the effective employment of substitute drugs in the current scenario of dwindling official botanical sources for many Traditional Siddha formulations.

Antioxidant Role of Kaempferol in Prevention of Hepatocellular Carcinoma.

Reactive oxygen species (ROS) are noxious to cells because their increased level interacts with the body’s defense mechanism. These species also cause mutations and uncontrolled cell division, resulting in oxidative stress (OS). Prolonged oxidative stress is responsible for incorrect protein folding in the endoplasmic reticulum (ER), causing a stressful condition, ER stress. These cellular stresses (oxidative stress and ER stress) are well-recognized biological factors that play a prominent role in the progression of hepatocellular carcinoma (HCC). HCC is a critical global health problem and the third leading cause of cancer-related mortality. The application of anti-oxidants from herbal sources significantly reduces oxidative stress. Kaempferol (KP) is a naturally occurring, aglycone dietary flavonoid that is present in various plants (Crocus sativus, Coccinia grandis, Euphorbia pekinensis, varieties of Aloe vera, etc.) It is capable of interacting with pleiotropic proteins of the human body. Efforts are in progress to develop KP as a potential candidate to prevent HCC with no adverse effects. This review emphasizes the molecular mechanism of KP for treating HCC, targeting oxidative stress.

ACNPD: The Database for Elucidating the Relationships Between Natural Products, Compounds, Molecular Mechanisms, and Cancer Types.

Objectives: Cancer is well-known as a collection of diseases of uncontrolled proliferation of cells caused by mutated genes which are generated by external or internal factors. As the mechanisms of cancer have been constantly revealed, including cell cycle, proliferation, apoptosis and so on, a series of new emerging anti-cancer drugs acting on each stage have also been developed. It is worth noting that natural products are one of the important sources for the development of anti-cancer drugs. To the best of our knowledge, there is not any database summarizing the relationships between natural products, compounds, molecular mechanisms, and cancer types.Materials and methods: Based upon published literatures and other sources, we have constructed an anti-cancer natural product database (ACNPD) (http://www.acnpd-fu.com/). The database currently contains 521 compounds, which specifically refer to natural compounds derived from traditional Chinese medicine plants (derivatives are not considered herein). And, it includes 1,593 molecular mechanisms/signaling pathways, covering 10 common cancer types, such as breast cancer, lung cancer and cervical cancer.Results: Integrating existing data sources, we have obtained a large amount of information on natural anti-cancer products, including herbal sources, regulatory targets and signaling pathways. ACNPD is a valuable online resource that illustrates the complex pharmacological relationship between natural products and human cancers.Conclusion: In summary, ACNPD is crucial for better understanding of the relationships between traditional Chinese medicine (TCM) and cancer, which is not only conducive to expand the influence of TCM, but help to find more new anti-cancer drugs in the future.

Present Circumstances in Nutraceuticals: A Review

Nutraceuticals are biologically energetic phytochemicals that own health benefits. Nutraceuticals are considered to be fortified or enriched foods providing all the essential nutrients required for maintaining the most appropriate fitness. Nutraceuticals in vast are the food or a part of food gambling a vast function in enhancing and retaining everyday physiological function that continues healthy people. Nutraceuticals: on the basis in their herbal source, chemical grouping, categorized into 3 key terms-nutrients, herbal, dietary supplements. Nutraceuticals is used as a powerful instrument in preserving fitness and to behave in opposition to numerous sicknesses inclusive of obesity, cardiovascular sicknesses, cancer, osteoporosis, arthritis, diabetes, cholesterol and so forth. The goal of nutraceuticals in healthcare primarily based on their disease particular warning signs. In this paper we examine sources of nutraceutical and functional components in different foods and their assessment.

Authenticity control of pine sylvestris essential oil by chiral gas chromatographic analysis of \u03b1-pinene

Numerous terpenes present in essential oils (EOs) display one or more chiral centers. Within the same genus the enantiomeric ratio of these compounds can be different. Thus, the determination of enantiomers is a valuable tool to evaluate authenticity and quality of EOs. In here, the terpene profile of primary and commercial pine EOs was analyzed by conventional and chiral gas chromatography coupled to a flame ionization detector. The enantiomeric excess of ( ±)-α-pinene was determined and significant differences between primary and commercially available EOs were observed. Primary EOs of Pinus sylvestris L. showed a positive enantiomeric excess of (+)-α-pinene whereas commercial EOs labeled as P. sylvestris L. exhibited an enantiomeric excess of (−)-α-pinene. Thus, chiral analysis provides useful information on the authenticity of pine EOs and allows to uncover possible mislabeling, the use of the wrong herbal substance and sources of adulteration in pine oil.

Repurposing the inhibitors of COVID-19 key proteins through molecular docking approach

The severe acute respiratory syndrome coronavirus 2, famous as COVID-19, has recently emerged as a novel virus and imposed an unrecoverable loss to global health and the economy. At present, no effective drug against COVID-19 is available and currently available viral drugs targeting the viral key proteins of related RNA viruses have been found ineffective against COVID-19. This study evaluated the inhibitors of the viral proteases and other structural proteins, including Mpro (Main protease), RdRp (RNA-dependent RNA polymerase), and spike glycoprotein from synthetic and herbal sources. The molecular docking-based approach was used to identify and evaluate the putative inhibitors of key proteins involved in viral replication and survival. Furthermore, the pharmaceutical properties of these inhibitors were explored to predict the drug suitability as a therapeutic agent against COVID-19 by considering adsorption, distribution, metabolism, and excretion (ADME) using Lipinski’s rule or SwissADME. Trandolapril, Benazepril, and Moexipril were evaluated as the best non-carcinogenic and non-toxic potential inhibitors of spike glycoprotein, Mpro, and RdRp, respectively. The drugs showed significant binding affinities against the active sites of respective SARS_CoV-2 target proteins; hence, they can be used as potential therapeutic agents for the treatment of COVID-19.

Targeting miRNA by Natural Products: A Novel Therapeutic Approach for Nonalcoholic Fatty Liver.

The increasing prevalence of nonalcoholic fatty liver disease (NAFLD) as multifactorial chronic liver disease and the lack of a specific treatment have begun a new era in its treatment using gene expression changes and microRNAs. This study aimed to investigate the potential therapeutic effects of natural compounds in NAFLD by regulating miRNA expression. MicroRNAs play essential roles in regulating the cell's biological processes, such as apoptosis, migration, lipid metabolism, insulin resistance, and adipocyte differentiation, by controlling the posttranscriptional gene expression level. The impact of current NAFLD pharmacological management, including drug and biological therapies, is uncertain. In this context, various dietary fruits or medicinal herbal sources have received worldwide attention versus NAFLD development. Natural ingredients such as berberine, lychee pulp, grape seed, and rosemary possess protective and therapeutic effects against NAFLD by modifying the gene's expression and noncoding RNAs, especially miRNAs.

Chemical compositions, antibacterial, antifungal and cytotoxic effects of Alhagi mannifera five extracts.

Plants were used as medicines thousands of years ago. Conventional medicine use is increasing and many of the currently used drugs are extracted from herbal sources. In Palestinian traditional medicine, the Alhagi mannifera plant is used for the treatment of cancer. Our study aimed to extract this plant using five solvent fractions, identifying their chemical compositions, and evaluating their antimicrobial and cytotoxic effects. The successive technique was used to extract five solvent fractions of A.mannifera. While the spectral analysis was used to characterize quantitatively and qualitatively the chemical components of these extracts. The antimicrobial activity of plant extracts was evaluated against seven microbial strains using a broth micro-dilution assay. The cytotoxic activity was assessed using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay against cervical cancer cell line (HeLa). A total of 165 compounds were identified in A.mannifera different extracts. In the petroleum etherextract were found a total of 55 compounds. The major compounds were 2,5-cyclooctadien-1-ol (9.42%), 3-chloropropionic acid, heptyl ester (9.42%), carbonic acid, ethyl nonyl ester (9.42%) and chloroacetic acid. In methylene chloride extract a total of 11 compounds were found, and the major compounds were m-ainobenzenesulfonyl fluoride (14.35%), dodecane,2,6,10-trimethyl- (14.35%) and propanoic acid,2,2-dimethyl-,2-ethylexyl ester (14.35%). In chloroform extract, a total of 23 compounds were found. The major compounds were 5-ethyl-1-nonene (21.28%), and decanedioic acid, bis(2-ethylhexyl) ester (21.28%). In acetone extract were found a total of 47 compounds and the major compound was phenol,2,4-bis(1,1-dimethylethyl)- (5.22%). In methanol extract a total of 29 compounds were found and the major compounds were 3-o-methyl-d-glucose (10.79%), myo-inositol, 2-c-methyl- (10.79%), myo-inositol, 4-c-methyl- (10.79%), and scyllo-inositol,1C-methyl- (10.79%). All extracts showed antimicrobial activity. However, the petroleum ether extract showed the most potent antimicrobial effect against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, MRSA, and Candida albicans with minimal inhibitory concentration (MIC) values of 1.25, 1.25, 6.25, 0.325, 6.25, and 1.56μg/mL, respectively. De facto, chloroform extract followed by ether extract displayed potential cytotoxic activity with IC50 values of 0.2 and 1.2mg/mL, respectively. A.mannifera was found to contain a variety of phytochemicals and its chloroform extract showed a potent cytotoxic effect on HeLa cancer cells. In addition, petroleum ether showed potent antimicrobial agents and these extracts look promising as drug candidates. Further in vivo investigations should be conducted to provide the basis for developing new cancer and microbial infections treatments.

Withania Somnifera Linn as Herbal Source of Diverse Pharmacological Potential - A Comprehensive Review

Ashwagandha, also known as Indian winter cherry, belonging to the Withania Somnifera Linn. (Solanaceae) family, (commonly available as Churna). Africa, the Mediterranean, and India are all home to this species. It is 30-50cm high and mainly found in the drier parts of India. Since it raises haemoglobin (red blood count) and hair melanin. Withania Somnifera is known as "Rasayana," which means "strong rejuvenator" in Ayurvedic jargon. In ethnomedicine, it is a well-known health food and herbal tonic that is used to treat cardiovascular diseases. It can be used as a single herb or as part of polyherbal or herbo-mineral formulations for humans. It is traditionally used to treat various diseases such as inflammation, asthma, dyspepsia, hypertension, rheumatism, tumor, anxiety, hemopoietic, antimicrobial, depression, immunomodulation, antiulcer, hepatoprotective activities, Alzheimer’s, Parkinson’s disease, rejuvenating properties and syphilis. It has wide variety of chemical constituents such as withanoloides, flavonoid, saponins, glycosides, alkaloids, tannins, steroids have been studied. Despite the fact that previous review articles on this plant have been written, this review article is provided to bring together all of the most recent knowledge on its phytochemical and pharmacological activities.

Phytochemical analysis using GC-FID, FPLC fingerprinting, antioxidant, antimicrobial, anti- inflammatory activities analysis of traditionally used Eucalyptus globulus essential oil

Eucalyptus globules is an widely distributed in tropical and subtropical regions. It has been widely used as folk medicine, and folk cosmetic owing to its antioxidant values. Despite its importance, phytochemical and pharmacological studies remain infancy. This study was aimed at extraction of essential oil by steam-distillation and evaluation of bioactive components, antioxidant, antimicrobial, anti-inflammatory activities along with analysis by UV-VIS, FT-IR and Fluorescent techniques. Fast protein liquid chromatography (FPLC) was used to confirm the presence of polyphenols. Different antioxidant activities like DPPH., ABTS.+, .OH, superoxide, nitric oxide and reducing power of the essential oil. Essential oil was analyzed by UV-VIS, FT-IR and Fluorescent techniques. In vitro antimicrobial activity was also monitored. FT-IR fingerprint qualitative analysis was performed using commercial standards. Considerable amount of flavonoids were detected in essential oil. Oil exhibited considerable scavenging activities of ABTS.+, .OH, superoxide, nitric oxide and reducing power. UV-VIS, FT-IR analysis revealed the presence of polyphenolics in essential oil. Fluorescent spectroscopy revealed the presence of fluorophores in essential oil. FPLC and FT-IR fingerprint analysis revealed the presence of bioactive constituents like rutin, tannic acid, vanillic acid and ascorbic acid in the essential oil. A strong anti-inflammatory activity of oil was observed using fluorescent spectroscopy. An appreciable in vitro antibacterial activity against gram-negative bacteria like Acetobacter aceti and Pseudomonas aeruginosa was detected. The data provides the scientific support to the use of essential oil from Eucalyptus globules as a potent herbal source of bioactive compounds possessing natural antioxidant activities in food and pharmaceutical industries.

Preclinical Study on the Hepatoprotective Effect of Pollen Extract of Pinus brutia Ten. (Red Pine) in Mice and Phenolic Acid Analysis

Many agents, including those from herbal sources, have been sought as preventives or cures for hepatotoxicity. The pollen of Pinus brutia Ten., known as red pine (Pinaceae), is used against liver diseases in Anatolian folk medicine. In the current study, pollen ethanol extract of P. brutia was investigated for its possible hepatoprotective activity using a mouse model of CCl4-induced hepatotoxicity. Swiss albino mice were divided into five groups, and extract-treated groups were compared with a silymarin-treated group as the reference. The extract was tested at 100, 200, and 300 mg/kg (b.w.). Phenolic acids were analyzed using high-performance column chromatography (HPLC) in the extracts as pollens are usually known to be rich in phenolics. Our data revealed that the extract displayed the best hepatoprotection at a dose of 100 mg/kg when compared with silymarin (Legalon®), the reference drug. HPLC analysis indicated presence of protocatechuic acid (0.176 mg/g extract), p-hydroxybenzoic acid (0.001 mg/g extract), vanillic acid (VA) (0.537 mg/g extract), syringic acid (0.050 mg/g extract), and tr-cinnamic acid (0.310 mg/g extract), while the major phenolic acid was VA. The outcomes of this study allow us to conclude that red pine pollen extract can serve as a promising hepatoprotective agent. Among the phenolic acids analyzed in the pollen extract, vanillic acid as the major one besides some other phenolic acids detected seems to be responsible for its remarkable hepatoprotective effect.

Classification and Identification of Essential Oils from Herbs and Fruits Based on a MOS Electronic-Nose Technology

The frequent occurrence of adulterated or counterfeit plant products sold in worldwide commercial markets has created the necessity to validate the authenticity of natural plant-derived palatable products, based on product-label composition, to certify pricing values and for regulatory quality control (QC). The necessity to confirm product authenticity before marketing has required the need for rapid-sensing, electronic devices capable of quickly evaluating plant product quality by easily measurable volatile (aroma) emissions. An experimental MAU-9 electronic nose (e-nose) system, containing a sensor array with 9 metal oxide semiconductor (MOS) gas sensors, was developed with capabilities to quickly identify and classify volatile essential oils derived from fruit and herbal edible-plant sources. The e-nose instrument was tested for efficacy to discriminate between different volatile essential oils present in gaseous emissions from purified sources of these natural food products. Several chemometric data-analysis methods, including pattern recognition algorithms, principal component analysis (PCA), and support vector machine (SVM) were utilized and compared. The classification accuracy of essential oils using PCA, LDA and QDA, and SVM methods was at or near 100%. The MAU-9 e-nose effectively distinguished between different purified essential oil aromas from herbal and fruit plant sources, based on unique e-nose sensor array responses to distinct, essential-oil specific mixtures of volatile organic compounds (VOCs).

Virtual screenings of the bioactive constituents of tea, prickly chaff, catechu, lemon, black pepper, and synthetic compounds with the main protease (Mpro) and human angiotensin-converting enzyme 2 (ACE 2) of SARS-CoV-2

BackgroundCOVID-19 has mutation capability, and there are no specific drug therapies that are available to fight or inhibit the proteins of this virus. The present study aims to investigate the binding affinity of the bioactive and synthetic compounds with the main protease (Mpro) enzymes and angiotensin-converting enzyme 2 (ACE 2) by computational approach. PASS prediction, pharmacokinetics, and toxicological properties prediction studies were performed through the Google PASS prediction and Swiss ADME/T website. Besides, molecular docking studies were accomplished by BIOVIA Discovery Studio 2020, UCSF Chimera, and PyRx autodock vina.ResultsThe docking scores were inferred and the selected compounds showed results varying from −3.2 to −9.8 (kcal/mol). Theaflavin scored the highest docking score to the 5REB, 6VW1, and 1R42 enzymes and showed the binding affinity as −6.3 kcal/mol, −9.8 kcal/mol, and −8.6 kcal/mol, respectively. Again, kaempferol showed the best binding affinity to the 7BQY (−7.1 kcal/mol) and 6Y2FB (−6.6 kcal/mol) enzymes. All the chemical constituents showed better probability in action in pass prediction analysis. Besides, no ligands (except theaflavin) have any conflict with Lipinski’s rules of five, which authorized the drug probability of these ligands.ConclusionTherefore, the selected compounds could be considered a potential herbal treatment source against SARS-CoV-2.

Effect of drying and composition ratio of herbal tea prepared from Clitoria ternatea L. and Ocimum sanctum L. on its antioxidant capacity

Lemon Basil (Ocimum sanctum L.) and Butterfly Pea (Clitoria ternatea L.) have been prepared as an herbal tea source and evaluated their secondary metabolites content and antioxidant values. The purpose of this study was to determine the effect of drying time and composition ratio of herbal tea on their antioxidant value. The experimental design of Factorial Completely Randomized Design with nine treatment combination and three replications was applied. The results were analysed by ANOVA and Tukey’s HSD test. The results showed that the treatment affected the antioxidant value with the F-value > F-table (hypothesis accepted) at the test level 0.01 with a value of 94.26 > 3.71. The best treatment is P2K3 by showing an antioxidant value of 38.61 ppm. A total of 37.01 ppm concentration of combined herbal tea samples as a whole can reduce 50% of the radical effects of DPPH compounds. The secondary metabolite compounds contained in combination herbal teas are alkaloids, flavonoids, terpenoids, and saponins.

A new and rapid analysis method for the most important herbal squalene source: Comparison of UV–visible, fluorescence, and FTIR techniques for the quantification of squalene in amaranth seed oil

A new method for the quantification of squalene in amaranth seed oil (ASO) is proposed. The spectroscopic techniques of ultraviolet–visible (UV–vis), fluorescence, and Fourier transform infrared (FTIR) were tested using partial least square (PLS) calibration models. Based on the results obtained from the reference chromatographic method, two ASO samples with the highest and lowest concentrations among totally 40 ASO samples were selected to prepare the calibration set (CS) (n = 20). All ASO samples were also kept as prediction set (PS) for the control of the predictive capacity of the chemometric model to be developed. To quantify the squalene compound in ASO, a totally of 26 PLS calibration models were tested using all spectroscopic techniques with various spectral pretreatments, and the best PLS model was selected by evaluating their statistical values of root mean square error of calibration (RMSEC), root mean square error of cross-validation (RMSECV), and R-square. The best calibration model was obtained from normal spectra in FTIR spectroscopy (4000–400 cm−1) with the lowest RMSEC of 0.0930, RMSECV of 0.1301, and the highest R-square of 0.9989 for calibration. The results of this study revealed that FTIR spectroscopy coupled with PLS regression could be used for fast, accurate, and environment-friendly quantification of squalene present in ASO samples.

Rational of Invertebrates and Herbs Extracts for Protection and Management of Coronavirus (COVID 19)

A novel outbreak of coronavirus [severe acute respiratory syndrome– coronavirus 2 (SARS-CoV-2)] has caused a global coronavirus disease pandemic in 2019 (COVID-19), leading to tens of thousands of infections and thousands of deaths worldwide. Coronaviruses are implicated in illnesses of humans, vertebrates and invertebrates. There is currently no effective treatment for COVID-19. Due to the high rate of transmission between humans of this virus and its pandemics, it is necessary to determine the fundamentals of its replication, structure and pathogenicity for finding a way to special treatment or prevention. COVID-19 virus pathogenesis has not been fully grasped. The key point of the pathogenic effect of COVID-19virus is its effect on angiotensin-converting enzyme2 (ACE2) receptors that play a major role in virus entry into the cell causing ending lesions. ACE2 receptors present in many human cell types in many organs such as lungs, arteries, heart, kidneys, intestines, nervous system and skeletal muscles. From ancient times, antiviral agents extracted from natural herbal and invertebrate sources have been used to prevent and treat a number of viral infections including respiratory viral infections. COVID-19 management is believed to be based on two strategies; enhancement of the immunity to control viral replication or virulence and relieve associated symptoms. This review strengthens the use of natural extracts as valuable tools to develop new therapeutic drugs against COVID-19 (Fig.1).

Evaluation of Anti-quorum Sensing Potential of Saraca asoca (Family Caesalpiniaceae) against Chromobacterium violaceum and Pseudomonas aeruginosa PA01

Aim: The present study was performed to evaluate the anti-quorum sensing (QS) potential of traditional medicinal herb Saracaasoca (family Caesalpiniaceae) stem bark extract against Chromobacterium violaceum and Pseudomonas aeruginosa PA01.
 Study Design: First, the test sample (bark extract) was screened for anti-QS activity. Then systematic in-vitro and biochemical tests were performed to evaluate the effect of the test sample on the QS mediated virulence factors.
 Place and Duration of Study: All the experimental works were performed in Lab 311, pharmacology division, CSIR-NBRI Lucknow from June 2019 to October 2019.
 Methodology: The samples of Saraca asoca stem bark were washed, dried and extracted using 70% methanol. The minimum inhibitory concentration (MIC) of the prepared Sarca asoca bark extract was determined using the Alamar blue assay, and the anti-QS activity was screened using standard agar overlay method against CV 12472 at subinhibitory concentrations 100, 200 and 300 µg (< MIC value). SAE effect on biofilms formation was assessed by growing biofilms on glass slides in a static culture of PA01. Anti-virulence effect of SAE on the production of QS-regulated virulence factors such as Pyocyanin, proteases, elastases, rhamnolipid and alginate in Pseudomonas aeruginosa was determined using the supernatant of a 24 hours old broth culture of PA01 supplemented with SAE. Using the agar plate technique, the swimming and swarming motility assays were conducted on 0.3% and 0.5% agar plates respectively. One-way ANOVA was used to analyze the data, presented as mean ± SD (standard deviation) of three independent experiments.
 Results: Preliminary screening results showed significant QS inhibition against CV 12472 in an agar overlay disk diffusion assay in a concentration-dependent manner. Data from the biofilm assay showed loose, distorted, irregular PA01 biofilm formation at 200 µg (48%) and 300 µg (65%). SAE caused a significant drop in virulence factor production, with maximum reduction in pyocyanin (58%), proteases (67%), elastases (52%), rhamnolipid (53%), and alginate (44%) observed at 300 µg concentration. At SAE sub-lethal concentrations (200 and 300 µg), both the swimming and swarming motility of PA01 were significantly inhibited.
 Conclusions: The present study demonstrates the broad-spectrum anti-QS potential of SAE, reported for the first time, suggesting that SAE could be considered as an alternative herbal source to develop antimicrobial agents which can be either solitary or synergistically with conventional antimicrobial drugs.

Evaluation of Analgesic Effect of Unani Pharmacopoeial Formulation Habb-E-Suranjan and Raughan-E-Suranjan in Wajaul Mafasil (Joints Pain) Patients: An Open Prospective Clinical Trial

Wajaul Mafasil (joint pain) is the most prevalent and common cause of morbidity and disability not only in India but also throughout the world. In spite of various scientific developments in conventional medicine, there is no specific therapy or cure available for joints pain particularly when the disease has progressed to chronic stage. Therefore, it is the need of the hour to explore new formulations from herbal sources which are safe and effective. The present trial was undertaken to scientifically validate the Unani pharmacopoeial compound formulations of Habb-e-Suranjan and Raughan-e-Suranjan for its safety and efficacy in patients with joints pain.
 Patients with joint pain, who gave written informed consent, were recruited in the study from general OPD of RRIUM, Chennai with the prior approval of the Institutional Ethics Committee. 97 patients completed the study treatment. The treatment protocol was followed for 14 days and the clinical evaluation was done on seventh and fourteenth day of the treatment. At the end of fourteenth day intensity of individual parameters like joints pain, swelling, tenderness and restriction of movement showed statistically significant improvement (p<0.05). Overall therapeutic response of the treatment observed in this study was 100%. It was observed that 47.4% cases were completely relieved; 51.4% cases were relieved while 1.2% cases showed partial relief after the treatment. The laboratory investigations showed that the drugs used in the study were safe as evident by its maintenance. It may be concluded that Habb-e-Suranjan in combination with Raughan-e-Suranjan possesses significant analgesic property and may be a better option for the patients of joint pain.
 Keywords: Arthritis, Joints pain, Wajaul Mafasil, Habb-e-Suranjan, Raughan-e-Suranjan

The antibacterial effect of β-pinene derived from Citrus aurantifolia against oral Streptococcus mutans

Introduction: Streptococcus mutans has been known to play a major role in dental caries development. This tooth decay is the most common oral disease affecting people in the world. Hence, discovering the new herbal antibacterial agent seems to become promising yet challenging. One of herbal antibacterial source is Jeruk Nipis or namely Citrus aurantifolia, as it is believed to contain biological active compounds that may act as antibacterial to kill pathogenic oral microbes. This study was aimed evaluated antibacterial effect of β-pinene derived from Citrus aurantifolia against oral Streptococcus mutans. Methods: Type of research is quasi experimental research. The essential of C. aurantifolia essential oil was prepared and isolated using the hydro-distillation technique and further isolation of antibacterial compounds was conducted by combination column chromatography using organic solvent, and the structure was determined by infrared (IR), 1D NMR of 1H-, 13C-NMR and DEPT 135° in CDCl3, and UV-Vis spectrometer methods. The antibacterial activity was tested against Streptococcus mutans using the Kirby-Bauer method. Results: 0.75% yield was extracted from the essential oil of Citrus aurantifolia and after purification, an antibacterial compound was identified as β-pinene with the molecular formula C10H16. Furthermore, the Citrus aurantifolia exerted inhibition growth of Streptococcus mutans at concentration of 2000, 1000, and 500 ppm showed reduction in paper disk as much as 13.0, 11.9, and 11.6 mm respectively. Conclusion: Antibacterial effect of β-pinene derived from Citrus aurantifolia against oral Streptococcus mutans. This is proven that β-pinene derived from Citrus aurantifolia prospective as antibacterial compound that potentially can be used as another herbal antibacterial of choice to manage dental disease associated with Streptococcus mutans infection.