scholarly journals Evaluation of the Antibacterial Efficacy of Azadirachta Indica, Commiphora Myrrha, Glycyrrhiza Glabra Against Enterococcus Faecalis using Real Time PCR

2016 ◽  
Vol 10 (1) ◽  
pp. 160-165 ◽  
Author(s):  
Suresh Anand ◽  
Mathan Rajan ◽  
Nagendrababu Venkateshbabu ◽  
Deivanayagam Kandaswamy ◽  
Yarramreddy Shravya ◽  
...  
Keyword(s):  
Real Time ◽  
Azadirachta Indica ◽  
Real Time Pcr ◽  
Negative Control ◽  
Glycyrrhiza Glabra ◽  
Antibacterial Efficacy ◽  
Neem Extract ◽  
Liquorice Extract ◽  
Group 5 ◽  
Group 2

Aim: To compare the antibacterial efficacy of Azadirachta indica (Neem), Commiphora myrrha (Myrrh), Glycyrrhiza glabra (Liquorice) with 2% Chlorhexidine (CHX) against E. faecalis by using Real Time PCR Materials and Methods: A total of fifty teeth specimens (n=50) were inoculated with E. faecalis for 21 days. Specimens were divided into five groups (Group 1: Myrrh, Group 2: Neem, Group 3: Liquorice, Group 4: 2% CHX and Group 5: Saline (negative control)). The intracanal medicaments were packed inside the tooth. After 5 days, the remaining microbial load was determined by using real time PCR Results: Threshold cycle (Ct) values of Myrrh extract, Neem extract, Liquorice Extract, 2% CHX and saline were found to be 30.94, 23.85, 21.38, 30.93 and 17.8 respectively Conclusion: Myrrh extract showed inhibition of E.faecalis equal to that of 2% CHX followed by Neem, Liquorice and Saline

Toxicity of bean cooking media containing EDTA in mice

2020 ◽  
Vol 36 (6) ◽  
pp. 436-445
Author(s):  
Sabry Ali El-Naggar ◽  
Karim Samy El-Said ◽  
Mona Elwan ◽  
Maysa Mobasher ◽  
Fotouh Mansour ◽  
...  
Keyword(s):  
Ethylenediaminetetraacetic Acid ◽  
White Blood Cells ◽  
Saline Group ◽  
Negative Control ◽  
Blood Levels ◽  
Group 4 ◽  
Liver And Kidney ◽  
Group 5 ◽  
Group 2 ◽  
Kidney Functions

The possible renal and hepatic toxicities of ethylenediaminetetraacetic acid (EDTA) in bean cooking media were studied using 100 male albino mice. Two sublethal doses of EDTA were used to explore their toxic effects; 20 mg/kg and 200 mg/kg, which corresponded to 1/100th and 1/10th of LD50, respectively. Accordingly, the toxicity study was performed using 50 mice, divided into five groups ( n = 10/group) as follows: group 1 (Gp1) served as a negative control and was orally administered normal saline; group 2 (Gp2) was administered the bean cooking medium; group 3 (Gp3) was administered EDTA (200 mg/kg); group 4 (Gp4) was administered bean cooking medium containing 20 mg/kg of EDTA; and group 5 (Gp5) was administered bean cooking medium containing 200 mg/kg of EDTA. The results showed no significant changes in liver and kidney functions in Gp2 while Gp3, Gp4, and Gp5 exhibited significant increases in adverse liver and kidney function markers. Hematocrit values were significantly decreased in Gp3 and Gp5, while the total white blood cells counts were significantly decreased in Gp3 and significantly increased in Gp5. The number of platelets was decreased in Gp3, Gp4, and Gp5. The blood levels of sodium (Na+), iron (Fe2+), and calcium (Ca2+) were decreased in Gp3, Gp4, and Gp5 due to the chelating effects of EDTA. The hepatic and renal architectures were disorganized in Gp3, Gp4, and Gp5 with some hemorrhagic manifestations in livers and kidneys of mice. These results demonstrate that EDTA in bean cooking is harmful in mice under the conditions of this study, and the potentially harmful effects in humans supports restricting its use.

scholarly journals Effects of sample handling on the detection of porcine reproductive and respiratory syndrome virus in oral fluids by reverse-transcription real-time PCR

2018 ◽  
Vol 30 (6) ◽  
pp. 807-812 ◽  
Author(s):  
Ashley C. Weiser ◽  
Korakrit Poonsuk ◽  
Sarah A. Bade ◽  
Phillip C. Gauger ◽  
Marisa Rotolo ◽  
...  
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
Real Time Pcr ◽  
False Negative ◽  
Rna Extraction ◽  
Free Water ◽  
Negative Control ◽  
Respiratory Syndrome Virus ◽  
Sample Handling ◽  
Oral Fluids

We evaluated effects of handling procedures on detection of porcine reproductive and respiratory syndrome virus (PRRSV) in oral fluids (OFs) by reverse-transcription real-time PCR (RT-rtPCR). The experiments were conducted using a composite sample of PRRSV-positive OF collected from 5-wk-old pigs vaccinated 15 d earlier with a modified-live PRRSV vaccine. Five pre-extraction sample-handling steps and all combinations thereof were evaluated: 1) thaw temperature (4°C or 25°C); 2) sample diluent (1:1 dilution with nuclease-free water or guanidinium thiocyanate–phenol); 3a) sonication of the sample (yes or no); 3b) temperature (4°C or 25°C) at which step 3a was conducted; and 4) temperature at which the sample was maintained after step 3b and until RNA extraction was initiated (4°C or 25°C). All combinations of the 5 sample-handling steps (i.e., 32 unique treatments) were tested in a completely randomized factorial design with 4 replicates and 1 negative control for each treatment. The entire experiment was repeated on 5 separate days to produce a total of 800 PRRSV RT-rtPCR results. Binary (positive or negative) data were analyzed by logistic regression and results (Ct) were analyzed using a generalized linear model. Overall, 1 false-positive result was observed among 160 negative controls (99.4% specificity), and 85 false-negative results were observed among the 640 known-positive samples (86.7% sensitivity). The most significant factor affecting test outcome was thaw temperature (4°C or 25°C); samples thawed at 4°C had higher positivity rate (94% vs. 80%, p < 0.0001) and lower Ct (36.2 vs. 37.5, p < 0.0001).

scholarly journals Evaluation of Mutagenic and Antimutagenic Activity of Methanol Extract of Cousinia thomsonii against Cyclophosphamide

2020 ◽  
pp. 53-57
Author(s):  
Ishfaq Shafi Khan ◽  
Khalid Bashir ◽  
Naveed Gulzar ◽  
Yaseen Maqbool Bhat
Keyword(s):  
Bone Marrow Cells ◽  
Positive Control ◽  
Negative Control ◽  
Male Rats ◽  
Free Access ◽  
Antimutagenic Activity ◽  
Male Wistar Rats ◽  
Group 5 ◽  
Group 2 ◽  
Group 1

The present study was conducted to evaluate the mutagenic and antimutagenic potential of Cousinia thomsonii (CT) extract in bone marrow cells of male wistar rats using some important parameters like micronucleated polychromatic erythrocyte (MnPCE), mitotic index (MI), chromosomal aberrations (CA) and polychromatic erythrocyte to normochromatic erythrocyte ratio (PCE/NCE). 30 male rats of wistar strain were divided into 6 groups with 5 rats each group. Group 1 rats were taken as negative control having free access to distilled water and rat feed. Group 2 rats were taken as positive control treated with mutagen cyclophosphamide (CP) at dose of 60 mg/kg b wt. for 2 days. Group 3 and 4 were treated with CT extract at dose of 100 and 200 mg/kg b wt. for 20 days. Group 5 and 6 were treated with 100 and 200 mg/kg b wt of CT extract for first 18 days and for last 2 days with CP at dose concentration of 60 mg/kg. It was found that rats treated with CT extract alone did not produce any significant changes in MnPCE, PCE/NCE ratio, CA and MI when compared with control treated rats (group 1). However in group 5 and 6 rats treated with CT extract in combination with CP a protective effect was observed against the cyclophosphamide induced cellular mutagenicity. In concluding remark Cousinia thomsonii was found to show antigenotoxic potential and also produce protective antimutagenic effects against CP induced chromosomal damage.

scholarly journals Antibacterial Efficacy of Casein Phosphopeptide-Amorphous Calcium Phosphate Compared to Calcium Hydroxide as Intracanal Medicaments against Enterococcus faecalis: In-vitro Study

2021 ◽  
Vol 15 (1) ◽  
pp. 257-261
Author(s):  
Rahaf Almohareb ◽  
Reem Barakat ◽  
Alhanouf Alshamsan ◽  
Manal Almutairi ◽  
Norah Alfuraih ◽  
...  
Keyword(s):  
Calcium Phosphate ◽  
Calcium Hydroxide ◽  
Enterococcus Faecalis ◽  
Amorphous Calcium Phosphate ◽  
Negative Control ◽  
Antibacterial Efficacy ◽  
Group 3 ◽  
Group 2 ◽  
Casein Phosphopeptide

Background: Endodontic infection may persist despite root canal instrumentation. Thus, the use of intracanal medicaments plays an essential role in eliminating resistant bacteria like Enterococcus faecalis, known to be associated with persistent infections in endodontically treated teeth. Although calcium hydroxide is the gold standard intracanal medicament, it has been reported that Enterococcus faecalis is immune to its effects. Therefore, several studies assessed the efficacy of other intracanal medicaments, but none to date evaluated Casein Phosphopeptide-Amorphous Calcium Phosphate. Objectives: This in-vitro randomized controlled study aimed to assess the antibacterial efficacy of Casein phosphopeptide-amorphous calcium phosphate as an intracanal medicament against Enterococcus faecalis and compared it to calcium hydroxide. Methods: 60 extracted single root canal permanent teeth were prepared and later divided into three equal groups according to the intracanal medicament used. Group 1: No intracanal medicament (negative control), Group 2: Calcium hydroxide paste, and Group 3: Casein phosphopeptide-amorphous calcium phosphate paste. The intracanal medicaments were placed on the canals for 7 days. The outcome of this procedure was measured by counting colony-forming units. Statistical analysis was carried out using One-Way ANOVA and Tukey’s Post Hoc Test to determine significant differences between the groups. Results: The mean bacterial count for Group 2 was significantly lower than Group 1 and Group 3. Calcium hydroxide showed significantly more antibacterial efficacy against Enterococcus faecalis than Casein phosphopeptide-amorphous calcium phosphate and the negative control groups. Conclusion: Casein Phosphopeptide-amorphous calcium phosphate is ineffective in inhibiting Enterococcus faecalis growth compared to Calcium hydroxide.

scholarly journals Detection of Salmonella typhimurium ATCC 14028 in Powder Prepared Traditional Medicines Using Real-Time PCR

2021 ◽  
Vol 4 (3) ◽  
pp. 178-183
Author(s):  
Alfi Sophian ◽  
Ratna Purwaningsih ◽  
Muindar Muindar ◽  
Eka Putri Juniarti Igirisa ◽  
Muhammad Luthfi Amirullah
Keyword(s):  
Salmonella Typhimurium ◽  
Real Time ◽  
Real Time Pcr ◽  
Positive Control ◽  
Negative Control ◽  
Pcr Analysis ◽  
Direct Pcr ◽  
Traditional Medicines ◽  
Pcr Method ◽  
Alternative Testing

The detection of Salmonella typhimurium ATCC 14028 using real-time PCR on powdered traditional medicinal products was carried out in the microbiology and molecular biology testing laboratory of the Food and Drug Administration in Gorontalo. This research aims to provide a reference for alternative testing methods in testing the products of traditional powder preparations on the market. The sample consisted of 10 traditional powder preparations spiked with positive control of S. typhimurium ATCC 14028 phase 2. The method used in the study was real-time PCR analysis using the SYBR® Green method, while DNA isolation using the direct PCR method. Data analysis was performed by analyzing the sample's melting temperature (Tm) curve and comparing it with positive control. The results showed that S. typhimurium ATCC 14028 was detected in samples at an average Tm value of 84.18°C, with ranges of 84.0-84.5°C. For positive control, the Tm value was at 85.2°C, while for the negative control, the Tm value was not detected. Based on these data, it can be concluded that S. typhimurium ATCC 14028 in traditional medicine products powder preparations can be detected using real-time PCR.

scholarly journals LAXATIVE EFFECT OF GREEN GEDI LEAVES INFUSES (Abelmoschus manihot (L.) Medik) ON MALE WHITE MICE (Mus musculus)

2019 ◽  
Vol 19 (2) ◽  
Author(s):  
Birgitta Vania Rarasati
Keyword(s):  
Experimental Study ◽  
Mus Musculus ◽  
Positive Control ◽  
Negative Control ◽  
P Value ◽  
Laxative Effect ◽  
Significant Difference ◽  
Group 5 ◽  
Group 2

Abstract. Background: Gedi plant (Abelmoschus manihot (L.) Medik) of the Malvaceae family is a plant which leaf is used by peoples for the treatment of several illnesses such as constipation. Purpose: To investigate the laxative activity of green gedi leaves infuses (Abelmoschus manihot (L.) Medik) on male white mice.Method: This study is an experimental study. Mice were divided in 5 groups of 6 animals each, first group as negative control (CMC Na 0.5%) while group 2, 3 and 4 were treated with green gedi leaves infuses (Abelmoschus manihot (L.) Medik) at doses of 130, 260 and 520 mg/kgBW, per as respectively and group 5 as positive control (bisacodyl). The laxative activity was determined based on the frequency of defecation, weight of feces and consistency of feces.Results: It is found that 520 mg/kgBW dose has the highest mean of defecating frequency (7.17) and 260 mg/kgBW dose has the highest mean of feces weight (0.20). Statically however, there is no significant difference between the groups overall with p value of 0.132 and 0.246 for defecating frequency and feces weight respectively. There is no difference between the groups in term of feces consistency with both not defecating and hard feces.Conclusion: The research concludes that there is no significant laxative activity between each groups after administration of several dosages in 6 hours on mice thus the use of green gedi leaves as laxative in society can not be proven empirically in the laboratory.Keywords: gedi leaf, Abelmoschus manihot (L.) Medik, laxative effect, constipation

Abstract 1412: Sonic Hedgehog Gene Transfer Promotes Angiogenesis and Myocardial Regional Blood Flow via PKC Dependent Netrin Expression

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Rafeeq P Ahmed ◽  
Husnain K Haider ◽  
Shujia Jiang ◽  
Muhammed R Afzal ◽  
Muhammed Ashraf
Keyword(s):  
Blood Flow ◽  
Stem Cell ◽  
Gene Delivery ◽  
Conditioned Medium ◽  
Real Time ◽  
Real Time Pcr ◽  
Regional Blood Flow ◽  
Control Groups ◽  
Group 3 ◽  
Group 2

Background: Hedgehog signaling effects endothelial and fibroblast cell migration and vascularization of various tissues during embryonic development. We propose here that stem cell based Shh gene delivery to the heart promotes neovascularization and enhances regional blood flow (RBF) in the infartced heart. Methods and Results: The pCMVShh plasmid was constructed and used for non-viral transfection of primary mesenchymal stem cell (MSC) culture. Successful transfection and expression of Shh in transfected MSCs ( Shh MSCs) was confirmed by immunoflourescence, real-time PCR and Western blotting. Real-time PCR based array showed upregulation of angiogenic factors in Shh MSCs including netrin (>62-fold) and iNOS (>100-fold). The expression of netrin and iNOS was PKC dependent and was abolished in the presence of 2.5 μ M chelerythrin. Transduction of iNOS gene into MSCs increased netrin expression implying a role for iNOS downstream of PKC. Western blot showed that 45-KDa PKM proteolytic subunit of PKC was involved in this activity. Shh MSCs conditioned medium was biologically active and caused greater migration and tube formation of human vascular endothelial in vitro in comparison with untransfected MSCs ( non-Shh MSCs) conditioned medium. For in vivo studies, 70 μ l DMEM without cells (group-1) or containing 1x10 6 male non-Shh MSC (group-2), or Shh MSCs (group-3) were transplanted into female rat model of acute coronary artery ligation. S ry -gene studies on day-4 after cell engraftment showed higher survival of Shh MSCs ( p < 0.05 vs non-Shh MSC). Immunostaining for vWillebrand Factor-VIII and smooth muscle actin showed robust angiogenesis with distinctly large vessels in group-3 ( p < 0.001 vs group-2). RBF measured by fluorescent microspheres was increased ( p < 0.01 ) and infarction size was attenuated ( p < 0.05 ) in group-3 as compared with control groups. Left ventricle ejection fraction (52.3±2.6%) and fractional shortening (21.8±1.2%) were more preserved in group-3 as compared with control groups. Conclusions: Ex-vivo Shh gene delivery improves angiogenesis and RBF in the infarcted heart to preserve global heart function through the upregulation of multiple angiogenic cytokines including iNOS and netrin.

scholarly journals Influence of the ultrasonic activation of irrigating solutions on sealer penetration into lateral root canals

2018 ◽  
Vol 66 (2) ◽  
pp. 117-121 ◽  
Author(s):  
Thalles Henrique Macedo BARBOSA ◽  
Markelane Santana SILVA ◽  
Daylana Pacheco da SILVA ◽  
Antônio Carlos Mendes de MOURA ◽  
Maria Ângela Arêa Leão FERRAZ ◽  
...  
Keyword(s):  
Lateral Root ◽  
Microscopic Analysis ◽  
Negative Control ◽  
Ultrasonic Activation ◽  
Root Canals ◽  
Analysis Group ◽  
Group 4 ◽  
Cement Penetration ◽  
Group 5 ◽  
Group 2

ABSTRACT Objective: This study analyzed the influence of the irrigating solutions ultrasonic activation on the obturator cement penetration into lateral root canals. Methods: Fifty maxillary molars (palatine root) were randomly assigned to 5 experimental groups (Group 1 (EDTA 17% + manual agitation for 5 minutes), Group 2 (EDTA 17% + ultrasonic activation for 15 seconds), Group 3 (NaOCl 1% + Ultrasonic activation for 15 seconds), Group 4 (EDTA 17% + ultrasonic activation for 15 seconds and NaOCl 1% + ultrasonic activation for 15 seconds) and Group 5 (negative control). The lateral root canals were made in the apical and middle third. The obturation occurred in a single session. The radiographic and microscopic analyzes were performed to evaluate the sealant cement penetration degree. Data were analyzed by the Mann-Whitney and Wilcoxon tests. Results: Apical and middle third radiographic analysis showed that Groups 2 and 4 presented better penetration of the sealant cement. In the microscopic analysis, Group 4 presented superior results in relation to the other groups in both thirds. In the apical third, radiographic and microscopic analyzes showed significant differences in the comparisons between Group 4 and Groups 5 (p = 0.019) and 3 (p = 0.023) and between Group 5 and Groups 2 (p = 0.012), 3 (P = 0.038) and 4 (p = 0.019), respectively. Conclusion: It was concluded that the ultrasonic activation of the NaOCl 1% + EDTA 17% irrigation solution provides greater penetration of the endodontic cement in lateral root canals.

scholarly journals Effect of Trigonelline in a Model of Apoptosis in Rat Retina

2021 ◽  
Author(s):  
Sevinç Arzu Postacı ◽  
Ülkü Çeliker ◽  
Nevin İlhan ◽  
Ferda Dağlı ◽  
Ali Dal
Keyword(s):  
Positive Control ◽  
Negative Control ◽  
Male Rats ◽  
Rat Retina ◽  
Significant Difference ◽  
Group 6 ◽  
Group 3 ◽  
Group 5 ◽  
Group 2 ◽  
Group 1

Abstract Purpose: To evaluate the efficacy of drop trigonelline and oral trigonelline (TG) treatment in a model of N-methyl D-aspartate (NMDA)-induced retinal apoptosis in rat retina. To compare with brimonidine tartrate (BT) drops with known retinal neuroprotective activity.Methods: 42 Wistar Albino male rats were randomly divided into 6 groups of 7 each. No action was applied to Group 1. Group 2 (negative control) was given intravitreal Phosphate Bufferd Saline (PBS) on the first day of the experiment and did not receive any treatment. Groups 3, 4, 5 and 6 were given intravitreal NMDA on the first day of the experiment. Group 3 (positive control) didn't receive post-injection treatment. For 21 days from the second day of the experiment, oral TG was given to group 4, TG drops were given group 5 and BT drops were given group 6. Histopathological and biochemical evaluations were performed in all groups.Results: Severe retinal degeneration was observed in group 3 compared to group 2 (p<0.001).There was no statistically significant difference between group 1 and group 2 (p>0.05). TUNEL, Brn3a and caspase3 staining in group 5 and group 6 were similar to group 2 (p>0.05). Group 5 and group 6 compared to group 3 were observed significant decrease in iNOS levels(p<0.05). Decreasing MDA levels and increasing SOD levels were detected in group 4,5,6 compared to group 3 (p<0.05).Conclusion: In our study, it was determined that TG drops showed similar retinal neuroprotective efficacy to BT drops.

scholarly journals Detection and differentiation of Newcastle disease virus and influenza virus by using duplex real-time PCR.

2013 ◽  
Vol 60 (3) ◽  
Author(s):  
Dawid Nidzworski ◽  
Edyta Wasilewska ◽  
Krzysztof Smietanka ◽  
Bogusław Szewczyk ◽  
Zenon Minta
Keyword(s):  
Influenza Virus ◽  
Newcastle Disease Virus ◽  
Real Time ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Real Time Pcr ◽  
Influenza A ◽  
Egg Production ◽  
Negative Control ◽  
Pcr Assay

Newcastle disease virus (NDV), member of the Paramyxoviridae family and avian influenza virus (AIV), member of the Orthomyxoviridae family, are two main avian pathogens causing serious economic problems in poultry health. Both are enveloped, single-stranded, negative-sense RNA viruses and cause similar symptoms, ranging from sub-clinical infections to severe diseases, including decrease in egg production, acute respiratory syndrome, and high mortality. Similar symptoms hinder the differentiation of infection with the two viruses by standard veterinary procedures like clinical examination or necropsy. To overcome this problem, we have developed a new duplex real-time PCR assay for the detection and differentiation of these two viruses. Eighteen NDV strains, fourteen AIV strains, and twelve other (negative control) strains viruses were isolated from allantoic fluids of specific pathogen-free (SPF), embryonated eggs. Four-weeks-old SPF chickens were co-infected with both viruses (NDV - LaSota and AIV - H7N1). Swabs from cloaca and trachea were collected and examined. The results obtained in this study show that by using duplex real-time PCR, it was possible to detect and distinguish both viruses within less than three hours and with high sensitivity, even in case a bird was co-infected. Additionally, the results show the applicability of the real-time PCR assay in laboratory practice for the identification and differentiation of Newcastle disease and influenza A viruses in birds.

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