International Journal of

Pharmaceutical and Clinical Research

e-ISSN: 0975 1556

p-ISSN 2820-2643

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1. Identification of Potent Lead Molecules for Furin Receptor Through HTVS and Molecular Docking
Harsha Vardhan P, Vikas Reddy V, Mourya Reddy Ch, Rathi Suganya P
Objective: Furin is the family of proprotein convertase and localized in Trans Golgi Network (TGN) and cell surface. It is the major processing enzyme of the secretory pathway. Furin inhibitors are known to be therapeutic agents since it is involved in viral and bacterial activation, alzimer’s disease and cancer. Furin is the family of proprotein convertase and localized in Trans Golgi Network (TGN) and cell surface. It is the major processing enzyme of the secretory pathway. Furin inhibitors are known to be therapeutic agents since it is involved in viral and bacterial activation, alzimer’s disease and cancer. In this study, High Throughput virtual screening is used to screen natural and synthetic compounds and the short listed compounds are taken for checking the ADMET property. Methodology: The crystal structure of furin protein target was obtained from RCSB protein data bank (PDB Id: 4RYD) and protein preparation is done. Ligand from Zinc database are downloaded and prepared and taken for HTVS. Then the short listed compounds were taken for studying the ADMET property. Results: ZINC19799372, ZINC20411483, ZINC20412114, ZINC20411616 has highest docking score when compared with other small molecules. The drug-likeness and pharmacokinetic properties and other descriptors such as molecular weight, H-bond donors, H-bond acceptors, logP (octanol/water) and human oral absorption for the screened hits were predicted by Qikprop. The selected screened compounds ZINC19799372, ZINC20411483, ZINC20412114, ZINC20411616 has satisfactory percentage of oral absorption of >80% and are known to be potential lead compounds. Conclusion: In this study, we have screened the dataset of 1000 natural compounds and predicted the drug-likeness properties to identify the potent inhibitors against furin, and were also able to visualize the interactions of protein-ligand complex.

2. Evaluation of TNF-Α Expression After Adipose-Derived Mesenchymal Stem Cells (ASCs) Treatment Combined with Freeze-Dried Amniotic Membrane Wrapping in Rat Sciatic Nerve Defect Model
Utomo P, Permatasari N, Fibrianto Y H, Widodo M A
Nerve regeneration presents a clinical challenge to biomedical engineering. Recently, adipose-derived stem cells (ASCs) are known as one of the strategies in tissue regeneration. ASC has good potential to differentiate into neurogenic cells with capability in producing neuronal markers. This study aimed to evaluate the role of application of ASCs on sciatic nerve healing in association with the expression of TNF-α. Thirty-six Spraque-Dawley rats weighing approximately 250 g were divided into 6 control group and 6 experimental groups (n:3). The 6 experimental group was divided into evaluation on the 1st,3rd,5th,7 th 14 th and 21 st day after surgery. ASCs was applied to the injury site (dose 1 x 106 cells/nerve) with the freeze-dried amniotic membrane. Histopathology was evaluated with Hematoxylin-eosin and Masson’s Trichrome staining. Application of ASCs could down-regulate the TNF-α expression, decrease inflammation and fibrous collagen formation during nerve healing. Moreover, it can conclude that ASCs with freeze-dried amniotic membrane wrapping can decrease the inflammation and perineural fibrous collagen formation during Sciatic nerve healing

3 Drug-Hormone Interference Affects Pituitary-Thyroid-Adrenal Axis in Urinary Tract Infection
Ahmed A Abd-Rabou, Hanaa H Ahmed, Emad F Eskander
Nitrofurantoin has been used commonly as an antimicrobial treatment for urinary tract infections. Although the combination of lung and liver cytotoxicity is very rare, it may occur up on treatment with nitrofurantoin. Nitrofurantoin-associated pulmonary interactions are common among females. The anticholinergic and antimuscarinic effects of Flavoxate hydrochloride enhance the selective relaxant activity of strong smooth muscle on the urinary tract to relief pain and vesicourethral spasms.  It used for urinary frequency and incontinence associated with the urinary tract inflammations. Although various studies indicated the presence of burden effects of Nitrofurantoin and Flavoxate, there is a lack of literature studied the impact of those drugs on pituitary-thyroid-adrenal axis in urinary tract infections. Blood samples were withdrawn via venipuncture from male and female patients at the end of the treatment period and healthy controls to measure serum Follicle-stimulating hormone (FSH), Estradiol (E2), Luteinizing hormone (LH), Prolactin, Progesterone, Dehydroepiandrosterone (DHEAS), Testosterone, Thyroid-stimulating hormone (TSH), Triiodothyronine (T3), Thyroxine (T4), Fasting Insulin, and Cortisol. The circulating hormonal levels were measured quantitatively using Enzyme-link immunosorbent assay (ELISA). Hormonal profiling showed that there were remarkable signatures could be of great interest to underline some recommendations and guidelines optimizing the drug dosage to avoid burdens associated with those drugs as well as maintain the physiological and psychological performances of both sexes.

4 Altered Acetylcholinesterase and Na+-K+ATPase Activities in Different Areas of Brain in Relation to Thyroid Gland Function and Morphology Under the Influence of Excess Iodine
Jagadis Mandal, Arijit Chakraborty, Amar K Chandra
Background and objectives: Thyroid hormone exercises vital role in the event of development of central nervous system in stage of fetal life, childhood, and maintains tropic action in adulthood and also in aging. Deficiency in thyroid hormones responsible for impairment of mental performance. Inadequate iodine intake was is the major health problem. Use of iodized salt is now in vogue globally including India to eradicate iodine deficiency disorders (IDDs). This salt iodization should be at par to the iodine content of environment or water. Hilly areas have low iodine content whereas in coastal areas have higher iodine content in vegetables and water; however the people of both the areas are consuming uniform iodine containing salt. Therefore there is a chance of consuming excess iodine of the people in environmentally iodine sufficient areas. So far the effect of excess iodine specially in brain in relation to thyroid gland function yet have not been extensively investigated. Materials and method: For the study 36 number of rats weighing 120±10g of 90±5 days old, primarily divided into two groups on the basis of the duration of treatment viz. 30 days group and 60 days group respectively and on the exposure of the dose of iodine was divided in to two sub-group, excess iodine exposed group (100HI), was gavaged orally potassium iodide (KI) at the dose of 7 mg/Kg body weight and excessive iodine group (500HI) was gavaged orally KI at the dose of 35mg/Kg body weight. Results: Our overall result revel that those doses of excessive iodine causes a state of hypothyroidism as manifested by serum T3 serum T4 and TSH levels and thyroid morphology and also alters of cholinergic homeostasis in different areas of brain as revealed by acetylcholinesterase and Na+-K+ ATPase activity. Conclusion: Excessive iodine with higher dose and duration causes hypothyroidism which is responsible for alteration of cholinergic homeostasis in different region of brain.

5 Enhanced Blood–Brain Barrier Transmigration Using the Novel Chrysin Embedded Solid Lipid Nanoformulation: A Salient Approach on Physico-Chemical Characterization, Pharmacokinetics and Biodistribution Studies
Aishwarya V, Sumathi T
Solid lipid nanoparticles are an alternative carrier system used to load the drug for targeting, to improve the bioavailability by increasing its solubility, and protecting the drug from presystemic metabolism. The present study aims at determining the physico-chemical characteristics, stability, in-vivo plasma pharmacokinetic and bio-distribution analysis of chrysin loaded solid lipid nanoparticles (CN-SLNs) for exploring the targeted brain delivery. FTIR spectra of CN-SLNs exhibited a possible chemical interaction between chrysin and the polymer matrix having the shift in the absorption peaks of C-O stretch and “oop” band C-H bend. Differential Scanning Calorimetric study of CN-SLNs showed the complete disappearance of the characteristic peak of chrysin which reveals that the drug is molecularly dispersed into the lipid. The stability study of CN-SLNs indicates the stability of the nanoformulations without changing its performance on storage for 6 months. Furthermore, the in-vivo pharmacokinetic study revealed that encapsulation of chrysin in solid lipid nanoparticles increased the oral bioavailability of chrysin to 5 folds when compared with that of free chrysin. In biodistribution studies, CN-SLNs could be detected in the evaluated organs, including liver, heart, spleen, lung, kidney and brain. Solid lipid nanoformulation significantly raised the chrysin concentration especially in the brain which emphasizes on the blood–brain barrier permeation. These findings thus provide further understanding for the possible therapeutic effects of CN-SLNs in further brain disorder related clinical research.

6 Antibiotic Resistance of Staphylococci Concerning Strains Included in Food Industry in Egypt
Arafa A A, Ibrahim E S, Fouad E A, Gaber E S
Antibiotic resistance increased in humans and in veterinary medicine which may occur from direct way like improper treatment with antibiotic or addition of sub therapeutic doses of antibiotic in ration but there is also an in direct way which is food product as it may contaminated with multiple antibiotic resistance bacteria or bacteria carried the antibiotic resistance genes even if not expressed as we detect in our study so the objective of this study is to isolate and identify staphylococci from raw cow milk, pasteurized milk, pasteurized yogurt, beef burger and minced beef meat and to examine the isolated staphylococci phenotypically against seven 7 anti-microbial agents considered the most important agents: ciprofloxacin (5 μg), erythromycin(15 μg), tetracycline (30 μg), methicillin (5 μg), rifampicin (5 μg), penicillin (10 IU) and vancomycin (30 μg). We investigated the presence of antibiotic resistance genes by PCR; the β-lactamase gene blaZ(PEN resistance),mecA (methicillin resistance) and the tetK (TET resistance) that confer resistance to tetracycline. In the present study we record S. aureus and coagulase negative staphylococci (CNS) with multiple antibiotic resistance attributes. Also we found isolates of S. aureus and CNS carried blaZ gene and mecA gene but not expressed when they examined by disc diffusion test which let us highlight on these isolates as it could be a source of transmission of resistant genes to human and other bacteria. We also for the first time record S. aureus with 100% resistance to tetracycline recovered from food product phenotypically and genotypically while resistance to tetracycline was 33.3% in S. epidermidis.

7 Antiobesity Effect of Benincasa hispida Fruit Extract in High Fat Diet Fed Wistar Albino Rats
Nadhiya K, Vijayalakshmi K, Gaddam Aadinath Reddy G
Aim: The study was designed to evaluate the efficacy of ethanolic extract of Benincasa hispida (EEBH) and its active fraction (AFBH) on high fat diet fed rats. Methods: Male wistar rats (200±10g) were divided into five groups of 6 rats. Group I (Normal control), Group II (High fat diet(HFD), Group III (HFD+300mg/kg ethanolic fruit extract), Group IV (HFD+100mg/kg active fraction from ethanolic fruit extract) and Group V (HFD+25mg/kg Orlistat (drug control). After induction of High fat diet, Physical parameters such as body weight, organ weight, fat pad weight, anthropometric parameters were measured. Biochemical parameters such as serum lipid profile, Glucose, insulin, Homeostatic insulin resistance (HOMOIR), Free fatty acids, Phospholipids, homocysteine, apolipoprotein-B(Apo-B), leptin and adiponectin levels were analysed. Enzyme parameters such as Aspartate transaminase (AST), Alanine transaminase (ALT), Alkaline phosphatase (ALP), Lactate dehydrogenase (LDH), Creatinine kinase (CK) and lipase were analysed. Histopathological changes in the adipose tissue were identified using haemotoxylin eosin stain and similarly histopathological changes in the liver were identified using haemotoxylin eosin stain and oil red O staining. Results: Active fraction of Benincasa hispida showed better result in reducing lipid levels such as cholesterol and Triglycerides (TG), Low density lipoprotein (LDL), free fatty acids, Phospholipids, Insulin, HOMO-IR, leptin and enzyme levels. Homocysteine and apolipoprotein B (Apo-B) was an important cardioprotective marker. These parameters also were reduced in AFBH and EEBH treated groups, EEBH and AFBH increases HDL and adiponectin levels, when compared with HFD fed groups.  Antilipidemic activity was observed with AFBH.

8 Fas-Ligand and Granzyme-b Levels in Children with Nephrotic Syndrome
Gamila S M Elsaeed, Manal F Elshamaa, Doaa M Salah, Enas Mokhtar, Tarek Farid, Eman A Elghoroury
There is an urgent need to develop alternative to kidney biopsy in children to diagnose nephrotic syndrome. Increased apoptosis plays a central role in the development of nephrotic syndrome. The aim of this study was to evaluate the blood levels of two markers of apoptosis, Fas-ligand (Fas-l) and granzyme-b in children with nephrotic syndrome. Thirty children with biopsy-proven nephrotic syndrome as well as twenty four healthy controls were evaluated in this study. Clinical and laboratory data of included cases and controls were collected. Serum Fas-ligand and granzyme-b levels were measured using specific enzyme-linked immunosorbent assay (ELISA). The results of the current study showed that children with nephrotic syndrome had significantly higher serum levels of Fas-ligand (183.29±43.34pg/ml) and granzyme-b (151.68±30.47pg/ml) than healthy control subjects (Fas-1=99.28±7.20pg/ml, and granzyme-b=86.10±5.68pg/ml) (P=0.000). A significant increase in Fas-ligand levels in Focal segmental glomerulosclerosis cases than that of minimal changes nephrotic syndrome cases (p=0.004) was found. Granzyme-b levels were not significantly differing between the two groups of nephrotic syndrome cases.  In conclusion, markers of the extrinsic pathway of kidney apoptosis as Fas-ligand and granzyme-b are potential novel biomarkers in children with nephrotic syndrome that could be used as non invasive alternative to kidney biopsy in assessment of those children.

9 Synthesis and Molecular Docking Study of 4-Chlorophenylquinazoline-4-[3h]-One Derivatives as COX-2 Inhibitor
Tutuk Budiati, Suko Hardjono, Melanny Ika Sulistyowaty
According to data from the World Health Organization in 2014, cancer was the second biggest cause of death after heart disease. Several attempts have been made to produce the anticancer drug candidate. Modification of molecules against cancer drugs have been done so they became more effective and efficient. One mechanism of action of cancer drugs is to inhibit COX-2. Quinazolinone derivative compounds has anticancer activity, so this study synthesized some phenylquinazolinone derivatives. Virtual screening was carried out through docking the design compounds into the binding site of COX-2 enzyme (PDB code 3LN1) to predict if these compounds had analogous binding mode to the COX-2 inhibitor. Results obtained in the form of bond energy, indicated by the value of RS. Novel of phenylquinazolinone (4a-h) have been synthesized using anthranilic acid as starting material in three steps reaction. The purity of synthesized compounds was tested by TLC and m.p. data. The structures of the synthesized compound were identified using UV, IR and 1H-NMR spectra. The small RS value indicated a molecular bond that was stable and predictable had high activity. The smallest RS value was -122.54 kcal / mol and synthesized ranged between 70-85%.

10 Formulation and Evaluation of Moxifloxacin Ocusert
Gadhave M V, Pawar Sagar Dhondiram, Khilari Shobha Sopan, Gaikwad D D, Jadhav S J, Vjsm’s
Moxifloxacin,an antibacterial is effective against bacterial conjunctivitis and is new fourth generation of fluoroquinolone antibacterial drug. Ocuserts prepare by using different combination of polymer for better ocular drug delivery and increased retention time. In which ocusters have shown advantages to overcome disadvantages of conventional dosage form. Conventional ophthalmic formulations like solution, suspension, and ointment have many disadvantages which result into poor bioavailability of drug in the ocular cavity. The ocuserts were prepared by solvent casting technique. ocusert were characterized using different analytical techniques UV spectroscopy, FTIR, DSC. The drug content of the ocusert was found to be varied between 96.00 – 97.90 %. Ocusert provide better in vitro controlled release i.e. 77.13%was found to be better than the other formulations over an extended period of 8 hours and showed excellent microbiological activity for 3 days and remained stable and intact at ambient conditions.

11 An Efficient Encapsulation of Thymoquinone Using Solid Lipid Nanoparticle for Brain Targeted Drug Delivery: Physicochemical Characterization, Pharmacokinetics and Bio-Distribution Studies
Surekha R, Sumathi T
The present study was framed to determine the physicochemical characteristics, stability, in-vivo plasma pharmacokinetics and bio-distribution analysis of thymoquinone encapsulated solid lipid nanoparticle (TQ-SLNs) for the brain targeted drug delivery. Fourier transform infrared spectroscopy (FTIR) of TQ-SLNs revealed a possible chemical interaction between the drug and the lipid molecules by showing a shift in the absorption peak of C-H in-plane bend and “oop” band C-H bend stretch of thymoquinone in the lipid core. The thermal behavior of TQ-SLNs revealed a melting enthalpy value of ∆H=131.7 J/g and attained peak at 69.5°C that found to have a greater stability and reduced lipid crystal matrices verifies efficient drug incorporation. Further, the stability analysis of TQ-SLNs showed a stable encapsulation enhancement effect for 6 months. In-addition, the in-vivo pharmacokinetic study showed nearly a 5-fold increase in the bioavailability of TQ-SLNs after orally administered to the animals. Further, the drug distribution analysis of TQ-SLNs was found to accumulate more in the brain than other organs, hence suitable for the brain targeted drug delivery.

12 Chemical Constituents of Flacourtia rukam Zoli. & Moritzi Fruit
Consolacion Y Ragasa, Jo Madeleine Ann Reyes, Theresa Joyce Tabin, Maria Carmen S Tan, Irving D Chiong, Robert Brkljača, Sylvia Urban
Chemical investigation of the dichloromethane extract of Flacourtia rukam Zoli. & Moritzi (Syn.  Flacourtia euphlebia Merr.) led to the isolation of monogalactosyl diacylglycerols (1), β-sitosteryl-3β-glucopyranoside-6β-O-fatty acid esters (2), β-sitosterol (3) and triacylglycerols (4) from the pulp; 3 and chlrorophyll a (5) from the fruit peel; and 4 from the seeds. The structures of 15 were identified by comparison of their NMR data with literature data

13 Impact of Tobacco Smoking on Renal Function Tests in Asymptomatic Individuals
Fateheya M Metwally, Asmaa Mahmoud Mohammed
Objectives of this work was studying the effect of smoking on renal function tests among apparently healthy smokers. Study design: This is a cross sectional study was implemented on 53 active tobacco smokers and 72 nonsmokers (control group). Methods: we studied the effect of smoking on renal functions using both conventional renal function tests namely; serum urea, serum creatinine, estimated glomerular filtration rate and one of the urinary early renal biomarkers namely; N acetyl-B-D glucosaminidase as an indicator for renal impairment. The results revealed statistically significant higher activity of the Urinary N-Acetyl-B-D- glucosaminidase activity (NAG index), (P-value=0.001) among smokers when compared to their matched nonsmokers. The proportion of smokers having elevated NAG activity (45.2%) was significantly higher than that among non-smokers (18.4%) with statistically significant difference (p-value=0.01). Conclusion: Results of this study revealed detection of early renal effects among asymptomatic tobacco smokers which couldn’t be detected by routine kidney function tests.

14 Effect of Progressive Muscle Relaxation and Aerobic Exercise on Anxiety, Sleep Quality, and Fatigue in Patients with Chronic Renal Failure Undergoing Hemodialysis
Elham Amini, Iraj Goudarzi, Reza Masoudi, Ali Ahmadi, Ali Momeni
Insomnia, anxiety, and fatigue are more common in hemodialysis patients than in healthy people and affect patients’ quality of life. In the present study, the ‏ effects of progressive muscle relaxation (PMR) and aerobic exercise on anxiety, sleep quality, and fatigue in patients with chronic renal failure undergoing hemodialysis were evaluated. In this double-blind clinical trial, 100 hemodialysis patients were randomly assigned to three groups: PMR, aerobic exercise, and control. Patients performed relaxation and aerobic exercise daily for 60 days. Questionnaires of anxiety, sleep quality, and fatigue were completed by participants before and after the interventions. Data were analyzed by Stata software. PMR program significantly decreased general anxiety, trait anxiety, state anxiety, and Beck anxiety and aerobic exercise significantly reduced beck anxiety. PMR program and aerobic exercise both significantly improved sleep quality in hemodialysis patients. PMR program significantly reduced Rhoten fatigue but did not affect Piper fatigue. Aerobic exercise had no effect on Rhoten and Piper fatigue. Results showed better function of PMR compared to aerobic exercise in improving the symptoms of anxiety, sleep disorders, and fatigue in hemodialysis patients. Given that fatigue and sleep quality cause severe anxiety and somehow undermine quality of life in hemodialysis patients, taking into account non-pharmacological treatments such as aerobic exercise particularly PMR is a highly economical but efficient and efficacious strategy to manage several problems of these patients. Healthcare teams can incorporate these safe programs in care designs.

15 Comparative Medication Histories Between Newly Hospitalized Hypertensive Patients and Hypertensive Outpatients in Cote D’Ivoire
Danho Pascal Abrogoua, Annick Grah, Armand Angely Koffi, Elisée Doffou, Beugré Kouassi, Toussaint Toutou
Introduction: The aim of medication history is to specify behaviors that can influence treatment and it is part of an aid to educational diagnosis and medication reconciliation. The objective of our study was to identify factors that may contribute to hospitalization, poor therapeutic response or the occurrence of drugs adverse events in newly hospitalized hypertensive patients (NHP) and hypertensive outpatients (HOP). Methods: A comparative descriptive cross-sectional study was carried out from January 2014 to June 2015 in hospitalization units (department of internal medicine at teaching hospital of Treichville and department of neurology at teaching hospital of Cocody) and ambulatory follow-up unit (medical consultation department at Heart Institute of Abidjan) with hypertensive patients in Cote d’Ivoire. A semi-directive interview was conducted with a questionnaire of medication history with the following factors: factors related to the understanding of treatment (FUT), factors related to the monitoring of the treatment (FMT), factors related to dietetic-hygienic measures (FDHM), factors related to the effects of treatment (FET), factors related to patients (FP). Results: A total of 50 NHP and 100 HOP were recruited. For FMT, regular monitoring by a physician [NHP (52%) vs HOP (64%); p= 0.00006)], occasional discontinuation of medication [NHP (54%) vs HOP (27%); p = 0.001], satisfaction with the mode and forms of drug administration [NHP (72%) vs HOP (88%); p = 0.014], monitoring by several doctors [NHP (24%) vs HOP (0%); p = 0.001] and the frequency of biological assessments [NHP (34%) vs HOP (87%); p= 0.001] differed significantly between both groups. Among factors related to hygienic-dietetic measures, only the presence of stress and anxiety differed significantly [NHP (88%) vs HOP (72%), p = 0.02]. For factors related to the effects of treatment, the perception of adverse drug effects greater than beneficial effects differed significantly [NHP (2%) vs HOP (11%); p = 0.047]. Allergy to certain drugs [NHP (12%) vs HOP (33%); p = 0.0057] is a factor related to patients that differed significantly between both groups. However, most of FUT, FP, FET and FDHM differed in a non-significant way between NHP and HOP. Conclusion: Factors related to NHP are elements to be considered in an educational program of HOP or on leaving hospital.

16 Isolation and Identification of Flavonoid Fractions from the Leaves of Volkameria inermis and its In-vitro Cytotoxic Study
Lavanya Krishnadhas, Santhi R, Annapurani
The current study aims to optimize the solvent for the isolation of flavonoid fractions and identification of flavonoid components through HPTLC and HPLC techniques and it’s in vitro cytotoxic study against Ehrlich Ascites Carcinoma (EAC) cell lines. In vitro cytotoxic assay tryphan blue dye exclusion was carried out against EAC cell lines. The results revealed that ethyl acetate was found as the best solvent for the isolation of flavonoid fractions and several kinds of flavonoids such as quercetin and kaempferol were found as the major flavonols in this plant. In vitro cytotoxic study suggests that the flavonoid fractions of this plant possess potent antitumor activity against EAC cell lines.

17 Comparative Evaluation of Antimicrobial Properties of Leaf Extracts of Achyranthes aspera Plant and Chlorohexidine Against Streptococcus mutans, Enterococcus faecalis and Whole Salivary Samples of Children in Mixed Dentition Age Group
Deshpande R R, Kontham U R, Shep S V, Gupta S L
The use of plants for healing purposes predates human history and forms the origin of much modern medicine. There are many well established medicinal plants which have been evaluated and are known to have significant therapeutic properties. However, antimicrobial efficacy of many other lesser known plants is yet to be verified. In this study, the ethanolic extract of leaves, Achyranthes aspera is evaluated for antimicrobial activity against test samples of Streptococcus mutans, Enterococcus faecalis and human saliva samples. The salivary samples were collected from children in mixed dentition age group with moderate caries activity. Antibacterial assay was carried out by assessing the growth inhibition potential of plant extracts against tested samples using agar diffusion method. The results were comparatively evaluated with Chlorhexidine using it as a standard chemotherapeutic agent. The results depict that the plant extracts have marked activity against the tested samples showing significant zones of growth inhibition. Thus, this in vitro study supports use of ethanolic leaf extracts of Achyranthes aspera as a preventive and therapeutic remedy for dental caries and its sequelae.

18 Secondary Metabolites from Ficus ampelas Burm.F.
Consolacion Y Ragasa, Maria Roxanne Macuha, Mariquit M De Los Reyes, Emelina H Mandia, Chien-Chang Shen
Chemical investigation of the dichloromethane extracts of Ficus ampelas Burm.f. has led to the isolation of squalene (1),  mixtures of β-amyrin fatty acid esters (2a) and α-amyrin fatty acid esters (2b) in a 1:2 ratio and β-sitosterol (3a) and stigmasterol (3b) in a 2:1 ratio, chlorophyll a (4), and saturated fatty acids (5) from the twigs; and 5, β-sitosteryl-3β-glucopyranoside-6β-O-fatty acid esters (6), and long-chain fatty alcohols (7) from the fruit. The structures of 17 were identified by comparison of their NMR data with those reported in the literature.

19 The Expression of TLR-2 and TLR-4 Protein in the Epithelial Cells of the Oral Mucosal Patients with Recurrent Apthous Stomatitis (RAS)
Diah Savitri Ernawati, Desiana Radithia, Bagus Soebadi, Aulanni’Am Aulanni’Am
Background: Toll-Like Receptors (TLRs) play an important role in the human immune system. The objective of this study was to disclose or discover RAS using etiopathogenetic molecular approach by observing the TLRs. Particular objective of this study was to understand the expression of toll-like receptors 2 (TLRs-2) and TLR-4 in epithelial cells of oral mucosa, and to investigate the role of toll-like receptors in the innate immunity. Methods: Human oral epithelial cells were obtained by scraping the oral mucosal from 40 patients with recurrent aphthous stomatitis and 10 healthy adult volunteers. The epithelial cells are made into smears. Immunohistochemistry was peformed for identification of TLR-2 and TLR-4 protein using monoclonal antibodies anti-TLR-2 and TLR-4. Result: TLR2 and TLR4 protein were expressed in the oral mucosal epithelial cells especially in surface and nuclear cells. The expression of the TLR-2 in patients with minor RAU was 41,02%, while major RAS is expressed 43,58%. RAS patients with positiv TLR-4 was 48,71% in major RAS, while in minor RAS TLR-4 was expressed 38,46%. Conclusion:This study is the first to establish the presence of both TLR-2 and TLR-4 protein on epithelial cells of oral mucosa, and their expression can be up-regulated in infectious conditions. These results show that TLR-2 and TLR-4 may play a important role in local host defense of oral mucosal.

20 Chemical Constituents of Melanolepis multiglandulosa (Reinw. Ex Blume)
Phelan G Apostol, Mariquit M De Los Reyes, Ian A Van Altena, Consolacion Y Ragasa
Chemical investigation of the dichloromethane extracts of Melanolepis multiglandulosa (Reinw. ex Blume) yielded taraxeryl fatty acid esters (1), squalene (2), (E)-3-alkenoic acids (3), β-carotene (4), a mixture of β-sitosterol (5a) and stigmasterol (5b), long-chain fatty alcohols (6), and long-chain hydrocarbons (7) from the leaves; and 7, triacylglycerols (8), and long-chain saturated fatty acid esters (9) from the twigs. The structures of 1-9 were identified by comparison of their NMR data with literature data.

21 Moderate Concentrations of TNF- α Induce BMP-2 Expression in Endothelial Cells
Oski Illiandri, Hidayat Sujuti, Nungki Permatasari, Setyawati Soeharto
Tumor necrosis factor-alpha (TNF-α) is best-known as a potent pro-inflammatory cytokine involved in many cardiovascular diseases. During vascular calcification, TNF-α has been reported topromote osteogenic differentiation of human vascular smooth muscle cells (VSMC) and mesenchymal stem cells (MSCs). In contrast, there is alack of data reporting the osteoinductive effect of TNF-α in endothelial cell. In this study, experiments were performed to investigate and determine theoptimum dose of TNF-α that induces expression of the osteoinductive factor bone morphogenetic protein (BMP-2) in endothelial cells. Human vein endothelial cells were treated with TNF-α at doses of 0, 2, 5, 10, or 20 ng/mL for2, 8, or 24 h time intervals. BMP-2 cell expression was evaluated using immunocytochemistry staining by calculating the percentage of BMP-2 positive cells. Apoptosiswas determined by counting the number ofpyknoticcells. In this study, we found that the optimum dose of TNF-α thatinduces BMP-2 expression in endothelial cells was 5 ng/mL at the 8 h time interval. Lower (2 ng/mL) orhigher (10 and 20 ng/mL) concentrations of TNF-α had minimal effects on BMP-2 expression. Moreover, higher concentrations of TNF-α treatment (10 and 20 ng/ml) at8 h and 24 h increased the presence of pyknotic endothelial cells, which representthefinal stage of apoptosis.

22 Antibacterial Activity of Several Plant Extracts Against Proteus species
Wurood Alwan Kadhim, Mohanad Jawad Kadhim, Imad Hadi Hameed
Infectious diseases are one of the major problems in developing as well as developed countries.  Plants produce a diverse range of bioactive molecules, making them rich source of different types of medicines. Most of the drugs today are obtained from natural sources or semi synthetic derivatives of natural products used in the traditional systems of medicine. Thus, it is a logical approach in drug discovery to screen traditional natural products. Approximately 20% of the plants found in the world have been submitted to pharmaceutical or biological test and a sustainable number of new antibiotics introduced in the market are obtained from natural or semi synthetic resources. Medicinal plants are finding their way into pharmaceuticals, cosmetics, and neutralceuticals. In pharmaceutical field medicinal plants are mostly used for the wide range of substances present in plants which have been used to treat chronic as well as infectious diseases. Long before mankind discovered the existence of microbes, the idea that certain plants had healing potential, indeed, that they contained what we would currently characterize as antimicrobial principles, was well accepted. Since antiquity, man has used plants to treat common infectious diseases and some of these traditional medicines are still included as part of the habitual treatment of various maladies. The drugs already in use to treat infectious disease are of concern because drug safety remains an enormous global issue. Most of the synthetic drugs cause side effects and also most of the microbes developed resistant against the synthetic drugs. To alleviate this problem, antimicrobial compounds from potential plants should be explored. These drugs from plants are less toxic, side effects are scanty and also cost effective. They are effective in the treatment of infectious diseases while simultaneously mitigating many of the side effects that are often associated with synthetic antimicrobials. Plants are rich source of antibacterial agents because they produce wide array of bioactive molecules, most of which probably evolved as chemical defense against predation or infection. A major part of the total population in developing countries still uses traditional folk medicine obtained from plant resources with an estimation of WHO that as many as 80% of world population living in rural areas rely on herbal traditional medicines as their primary health care, the study on properties and uses of medicinal plants are getting growing interests. In recent years, this interest to evaluate plants possessing antibacterial activity for various diseases is growing. The present antibacterial review of the plant extracts demonstrates that folk medicine can be as effective as modern medicine to combat pathogenic microorganisms. The millenarian use of these plants in folk medicine suggests that they represent an economic and safe alternative to treat infectious diseases.

23 Medicinal Plants Used for Treatment of Rheumatoid Arthritis: A Review
Mohanad Jawad Kadhim, Ashwak Falih Kaizal, Imad Hadi Hameed
The objective of this present review is to evaluate the therapeutic potential of Zingiber officinale in rheumatoid arthritis. We have also aimed to present a summary of mechanism of action of specific phytochemicals of Zingiber officinale to reduce the pain claimed by RA-affected patients. Rheumatoid arthritis (RA) is a chronic, inflammatory, autoimmune disease, which affects synovial tissue in multiple joints. Although conventional treatments of RA commonly alleviate the symptoms, high incidence of adverse reactions leads to research tendency towards complementary and alternative medicine. As various medicinal plants are traditionally used for the management of symptomatologies associated with RA in Persian medicine, we reviewed medicinal literature to confirm their efficacy in the management of RA. Key findings Scientific evidence revealed that traditional medicaments exert beneficial effects on RA through several cellular mechanisms including downregulation of pro-inflammatory cytokines such as TNF-a, IL-6 and NF-jB, suppression of oxidative stress, inhibition of cartilage degradation with destructive metalloproteinases and enhancement of antioxidant performance. Various active constituents from different chemical categories including flavonols, lignans, coumarins, terpenes, glycosylflavons, dihydroflavonols, phytoestrogens, sesquiterpene lactones, anthraquinones, alkaloids and thymoquinones have been isolated from the medicinal plants.

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