scholarly journals In Vitro Anti-Leptospiral Activity of Phyllanthus amarus Extracts and Their Combinations with Antibiotics

2021 ◽  
Vol 18 (6) ◽  
pp. 2834
Author(s):  
Che Ain Munirah Ismail ◽  
Zakuan Zainy Deris ◽  
Ruzilawati Abu Bakar ◽  
Nabilah Ismail
Keyword(s):  
Morphological Changes ◽  
Synergistic Effects ◽  
Microscopic Analysis ◽  
Modern Medicine ◽  
Sem Analysis ◽  
Penicillin G ◽  
Phyllanthus Amarus ◽  
Alternative Medicines ◽  
Leptospira Serovars

Despite modern medicine, there is an increasing trend for cases of the bacterial infection leptospirosis, and this has led to the exploration of alternative medicines from various sources including plants. The aim of this study was to investigate the in vitro anti-leptospiral activity of Phyllanthus amarus extracts alone and combined with penicillin G, ceftriaxone, and doxycycline. Antimicrobial susceptibility testing was performed using the microdilution broth technique upon methanol extract (ME), aqueous extract (AE), and antibiotics against the Leptospira interrogans serovars Australis, Bataviae, Canicola, and Javanica, to determine minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs). The results were analyzed using an ELISA microplate reader combined with microscopic analysis. Synergy testing using a checkerboard assay was performed to determine the fractional inhibitory concentration index values of extracts combined with antibiotics against leptospires. Scanning electron microscopy (SEM) was used to investigate morphological changes of leptospires caused by potential anti-leptospiral agents alone and combined with antibiotics. The MICs and MBCs for P. amarus extracts ranged from 100 to 400 µg/mL for AEs and from 400 to 800 µg/mL for MEs. Penicillin G was the most effective anti-leptospiral drug, with MICs and MBCs ranging from <0.01 to 0.78 and <0.01 to 3.13 µg/mL, respectively, followed by ceftriaxone, with both MICs and MBCs ranging from 0.05 to 0.78 µg/mL, and doxycycline, with MICs and MBCs ranging from 0.39 to 3.13 µg/mL and 12.5 to 25 µg/mL, respectively. Combinations of P. amarus extracts and antibiotics did not show synergistic effects on all tested Leptospira serovars, with some combinations demonstrating antagonistic effects. SEM analysis, however, showed distorted Leptospira surfaces. P. amarus AE performed better anti-leptospiral activity than P. amarus ME. The morphological effects of P. amarus extract alone and its combination with antibiotic on Leptospira cells revealed promising anti-leptospiral properties.

scholarly journals In Vitro Antileptospiral Activity of Trigona thoracia Propolis and its Synergistic Effects with Commonly Prescribed Antibiotics

2020 ◽  
Vol 19 (1) ◽  
Author(s):  
Siti Radziah Ismail ◽  
Salwani Ismail ◽  
Zakuan Zainy Deris ◽  
Nabilah Ismail
Keyword(s):  
Structural Changes ◽  
Inhibitory Concentration ◽  
Morphological Changes ◽  
Synergistic Effects ◽  
Antimicrobial Properties ◽  
Penicillin G ◽  
Pathogenic Leptospira ◽  
Leptospira Spp ◽  
Leptospira Serovars

Introduction: Trigona thoracica propolis is known to have antimicrobial properties, however its antileptospiral properties and its synergistic effects with commonly prescribed antibiotics are scarcely documented. This study aimed to evaluate the antileptospiral properties of Trigona thoracica against pathogenic Leptospira species (spp.) and to study its synergistic effects with commonly prescribed antibiotics. Materials and Methods: The tested Leptospira serovars were Australis, Bataviae, Canicola and Javanica. Aqueous extract propolis (AEP) and ethanolic extracts propolis (EEP) were used. Broth dilution methods were used to determine the Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and the synergistic effects between the propolis and the tested antibiotics. The synergistic effects was evaluated by using the fractional inhibitory concentration (FIC) index. Morphological changes of the treated Leptospira were observed under a Scanning Electron Microscope (SEM). Results: The AEP and EEP were found to have antileptospiral properties against the tested Leptospira spp. The synergy result showed that only combination of AEP and penicillin G against serovar Australis has demonstrated synergistic effect with the FIC index of 0.38. Morphological study using SEM showed significant structural changes of the treated Leptospira spp. Conclusions: The result suggests that Trigona thoracica propolis could potentially be used as either a complimentary or an alternative therapeutic agent against pathogenic Leptospira spp.

scholarly journals Isoalantolactone Enhances the Antimicrobial Activity of Penicillin G against Staphylococcus aureus by Inactivating β-Lactamase during Protein Translation

Pathogens ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 161 ◽  
Author(s):  
Yonglin Zhou ◽  
Yan Guo ◽  
Zhongmei Wen ◽  
Xinxin Ci ◽  
Lining Xia ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Inhibitory Concentration ◽  
Synergistic Effects ◽  
Protein Translation ◽  
Multidrug Resistant ◽  
Cd Spectroscopy ◽  
Penicillin G ◽  
Pcr Assay ◽  
Mechanism Of Inhibition

β-Lactamase-positive Staphylococcus aureus is one of the most prevalent multidrug-resistant pathogens worldwide and is associated with increasing threats to clinical therapeutics and public health. Here, we showed that isoalantolactone (IAL), in combination with penicillin G, exhibited significant synergism against 21 β-lactamase-positive S. aureus strains (including methicillin resistant S. aureus). An enzyme inhibition assay, a checkerboard minimum inhibitory concentration (MIC) assay, a growth curve assay, a time-killing assay, a RT-PCR assay and Circular Dichroism (CD) spectroscopy were performed on different β-lactamases or β-lactamase-positive S. aureus strains, in vitro, to confirm the mechanism of inhibition of β-lactamase and the synergistic effects of the combination of penicillin G and IAL. All the fractional inhibitory concentration (FIC) indices of penicillin G, in combination with IAL, against β-lactamase-positive S. aureus, were less than 0.5, and ranged from 0.10 ± 0.02 to 0.38 ± 0.17. The survival rate of S. aureus-infected mice increased significantly from 35.29% to 88.24% within 144 h following multiple compound therapy approaches. Unlike sulbactam, IAL inactivated β-lactamase during protein translation, and the therapeutic effect of combination therapy with IAL and penicillin G was equivalent to that of sulbactam with penicillin G. Collectively, our results indicated that IAL is a promising and leading drug that can be used to restore the antibacterial effect of β-lactam antibiotics such as penicillin G and to address the inevitable infection caused by β-lactamase-positive S. aureus.

Radiotherapy Reduces Microhardness and Mineral and Organic Composition, and Changes the Morphology of Primary Teeth: An in vitro Study

2018 ◽  
Vol 53 (3) ◽  
pp. 296-304 ◽  
Author(s):  
Lenita Marangoni-Lopes ◽  
Gabriela Rovai-Pavan ◽  
Carolina Steiner-Oliveira ◽  
Marinês Nobre-dos-Santos
Keyword(s):  
Primary Teeth ◽  
Morphological Changes ◽  
Artificial Saliva ◽  
Surface Hardness ◽  
In Vitro Study ◽  
Sem Analysis ◽  
Sem Images ◽  
Organic Composition ◽  
Nonnormal Distribution

Objective: We aimed to evaluate whether radiotherapy causes changes in the mineral composition, hardness, and morphology of enamel and dentin of primary teeth. Materials and Methods: Thirty specimens of primary teeth were subjected to radiotherapy. At baseline and after 1,080, 2,160, and 3,060 cGy, the specimens were subjected to microhardness, FT-Raman spectroscopy, and scanning electron microscopy (SEM) analysis. The pH of artificial saliva was determined, as were the calcium and phosphate concentrations. The data were subjected to the Shapiro-Wilk normality test, showed a nonnormal distribution, and were compared by the Kruskal-Wallis test. Results: The results showed that the microhardness of the enamel surface decreased after 2,160 cGy (281.5 ± 58 kgf/mm2) when compared to baseline (323.6 ± 59.5 kgf/mm2) (p = 0.045). For dentin, the surface hardness decreased after 1,080 cGy (34.9 ± 11.4 kgf/mm2) and 2,160 cGy (26 ± 3.5 kgf/mm2) when compared to baseline (56.5 ± 7.7 kgf/mm2) (p < 0.0001). The mineral and organic contents of phosphate (p < 0.0001), carbonate (p < 0.0001), amide (p = 0.0002), and hydrocarbons (p = 0.0031) of enamel decreased after 3,060 cGy (5,178 ± 1,082, 3,868 ± 524, 999 ± 180, and 959 ± 168 kgf/mm2, respectively). For dentin, we noticed a growing increase in phosphate v2, amide, and hydrocarbon content after 1,080 cGy (8,210 ± 2,599, 5,730 ± 1,818, and 6,118 ± 1,807 kgf/mm2, respectively) and 2,160 cGy (1,0071 ± 2,547, 7,746 ± 1,916, and 8,280 ± 2,079 kgf/mm2, respectively) and a reduction after 3,060 cGy (6,782 ± 2,175, 3,558 ± 1,884, and 3,565 ± 1,867 kgf/mm2, respectively) (p < 0.0001). SEM images showed cracks on enamel and degradation of peritubular dentin. Conclusion: We concluded that radiotherapy caused a reduction in surface hardness, changed mineral and organic composition, and promoted morphological changes on the enamel and dentin of primary teeth.

Microscopic Matrix Remodeling Precedes Endothelial Morphological Changes During Capillary Morphogenesis

2013 ◽  
Vol 135 (7) ◽  
Author(s):  
Claire McLeod ◽  
John Higgins ◽  
Yekaterina Miroshnikova ◽  
Rachel Liu ◽  
Aliesha Garrett ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Morphological Changes ◽  
Umbilical Vein ◽  
Microscopic Analysis ◽  
Matrix Remodeling ◽  
Collagen Gels ◽  
Surrounding Matrix ◽  
The Matrix ◽  
Umbilical Vein Endothelial Cells

The formation of microvascular networks (MVNs) is influenced by many aspects of the microenvironment, including soluble and insoluble biochemical factors and the biophysical properties of the surrounding matrix. It has also become clear that a dynamic and reciprocal interaction between the matrix and cells influences cell behavior. In particular, local matrix remodeling may play a role in driving cellular behaviors, such as MVN formation. In order to explore the role of matrix remodeling, an in vitro model of MVN formation involving suspending human umbilical vein endothelial cells within collagen hydrogels was used. The resulting cell and matrix morphology were microscopically observed and quantitative metrics of MVN formation and collagen gathering were applied to the resulting images. The macroscopic compaction of collagen gels correlates with the extent of MVN formation in gels of different stiffness values, with compaction preceding elongation leading to MVN formation. Furthermore, the microscopic analysis of collagen between cells at early timepoints demonstrates the alignment and gathering of collagen between individual adjacent cells. The results presented are consistent with the hypothesis that endothelial cells need to gather and align collagen between them as an early step in MVN formation.

scholarly journals Ligands of the Peripheral Benzodiazepine Receptor Are Potent Inhibitors of Plasmodium falciparum and Toxoplasma gondii In Vitro

2002 ◽  
Vol 46 (10) ◽  
pp. 3197-3207 ◽  
Author(s):  
Florence Dzierszinski ◽  
Alexandra Coppin ◽  
Marlene Mortuaire ◽  
Etienne Dewailly ◽  
Christian Slomianny ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Toxoplasma Gondii ◽  
Morphological Changes ◽  
Synergistic Effects ◽  
Benzodiazepine Receptor ◽  
Peripheral Benzodiazepine Receptor ◽  
Parasite Resistance ◽  
Antimalarial Agents ◽  
Parasite Replication

ABSTRACT The increase in resistance of the malaria parasite Plasmodium falciparum to currently available drugs demands the development of new antimalarial agents. In this quest, we have found that ligands to the peripheral benzodiazepine receptor such as flurazepam, an agonist of the benzodiazepine family, and PK11195, an antagonist derived from isoquinoline, were active against Plasmodium falciparum. These two compounds effectively and rapidly inhibited parasite growth in vitro, irrespective of parasite resistance to chloroquine and mefloquine. Treatment with both drugs induced a sharp and consistent decline in parasitemia, a complete inhibition of parasite replication, and the destruction of parasites within the host red blood cells. Using electron microscopy, we showed that dramatic morphological changes, involving swollen endoplasmic reticulum and the reduction of hemozoin, were consistent with parasite death. The potent activities of flurazepam and PK11195 were also evaluated for antagonist or synergistic effects with currently used antimalarial drugs such as chloroquine and mefloquine. Moreover, flurazepam was found to be active against Toxoplasma gondii, another member of the phylum Apicomplexa. Taken together, our results indicated that benzodiazepines could be considered promising candidates in the treatment of both malaria and toxoplasmosis.

scholarly journals Evaluation of Five Different Desensitizers: A Comparative Dentin Permeability and SEM Investigation In Vitro

2017 ◽  
Vol 11 (1) ◽  
pp. 15-33 ◽  
Author(s):  
Nasibe Aycan Yilmaz ◽  
Ertan Ertas ◽  
Hasan Orucoğlu
Keyword(s):  
Citric Acid ◽  
Morphological Changes ◽  
Artificial Saliva ◽  
Sem Analysis ◽  
Fluid Filtration ◽  
Dentin Permeability ◽  
Dentin Surface ◽  
The Difference ◽  
Self Etch

Background/Objective: The purpose of this study was to evaluate the efficacy and durability of five different dentin desensitizers (Gluma Desensitizer Powergel, Bifluorid 12, Gluma Self Etch Bond, D/Sense Crystal, Nupro Sensodyne Prophylaxis Paste with Novamin) on tubule occlusion and dentin permeability reduction in vitro. Method: The quantitative changes in permeability of 100 dentin discs were measured after desensitizer treatments and following post-treatments of 6% citric acid challenge for 1 min or immersion in artificial saliva for 24 hours under hydrostatic pressure generated by a computerised fluid filtration meter. Qualitative SEM analyses were also carried out. Results: Dentin permeability decreased after desensitizer application in all groups. Nevertheless, only the difference between ‘Gluma Self Etch Bond’ and ‘Nupro Sensodyne Prophylaxis Paste with Novamin’ groups was significantly different (p<0.05). Dentin permeability increased significantly after post-treatments (p<0.05). There was no statistically difference among the citric acid-subgroups (p>0.05). Of all the artificial saliva-subgroups, only the difference between ‘D/Sense Crystal’ and ‘Bifluorid 12’ was significantly different (p<0.05). In SEM analysis, morphological changes were detected on the dentin surface and within the tubules following desensitizer treatments and post-treatments. Conclusion: All the desensitizers significantly reduced dentin permeability by changing the morphology of the dentin surface and/or dentinal tubules. Following post-treatments, there was some reduction in the efficacy of the desensitizers which was represented by the reduction in permeability values. SEM analysis revealed some physical changes in the dentin structure which can partly give an explanation to the reduced efficacy of tested desensitizers.

scholarly journals Tensile bond strength and SEM analysis of enamel etched with Er:YAG laser and phosphoric acid: a comparative study In vitro

2008 ◽  
Vol 19 (1) ◽  
pp. 57-61 ◽  
Author(s):  
Luis H. Sasaki ◽  
Paulo D. C. Lobo ◽  
Yumi Moriyama ◽  
Ii-Sei Watanabe ◽  
Antonio B. Villaverde ◽  
...  
Keyword(s):  
Phosphoric Acid ◽  
Bond Strength ◽  
Er:Yag Laser ◽  
Morphological Changes ◽  
Sem Analysis ◽  
Tensile Bond Strength ◽  
Human Enamel ◽  
Morphological Differences ◽  
Phosphoric Acid Etching

Er:YAG laser has been studied as a potential tool for restorative dentistry due to its ability to selectively remove oral hard tissue with minimal or no thermal damage to the surrounding tissues. The purpose of this study was to evaluate in vitro the tensile bond strength (TBS) of an adhesive/composite resin system to human enamel surfaces treated with 37% phosphoric acid, Er:YAG laser (lambda=2.94 mum) with a total energy of 16 J (80 mJ/pulse, 2Hz, 200 pulses, 250 ms pulse width), and Er:YAG laser followed by phosphoric acid etching. Analysis of the treated surfaces was performed by scanning electron microscopy (SEM) to assess morphological differences among the groups. TBS means (in MPa) were as follows: Er:YAG laser + acid (11.7 MPa) > acid (8.2 MPa) > Er:YAG laser (6.1 MPa), with the group treated with laser+acid being significantly from the other groups (p=0.0006 and p= 0.00019, respectively). The groups treated with acid alone and laser alone were significantly different from each other (p=0.0003). The SEM analysis revealed morphological changes that corroborate the TBS results, suggesting that the differences in TBS means among the groups are related to the different etching patterns produced by each type of surface treatment. The findings of this study indicate that the association between Er:YAG laser and phosphoric acid can be used as a valuable resource to increase bond strength to laser-prepared enamel.

Correlation Between Cellular Functions and Morphological Changes of Human Trophoblast in Vitro

1975 ◽  
Vol 33 ◽  
pp. 600-601
Author(s):  
John C. Garancis ◽  
Robert O. Hussa ◽  
Michael T. Story ◽  
Donald Yorde ◽  
Roland A. Pattillo
Keyword(s):  
Electron Microscopy ◽  
Cellular Proliferation ◽  
Morphological Changes ◽  
Culture Fluid ◽  
Cellular Functions ◽  
Human Trophoblast ◽  
Transmission Electron ◽  
Marked Activity ◽  
Malignant Trophoblast

Human malignant trophoblast cells in continuous culture were incubated for 3 days in medium containing 1 mM N6-O2'-dibutyryl cyclic adenosine 3':5'-monophosphate (dibutyryl cyclic AMP) and 1 mM theophylline. The culture fluid was replenished daily. Stimulated cultures secreted many times more chorionic gonadotropin and estrogens than did control cultures in the absence of increased cellular proliferation. Scanning electron microscopy revealed remarkable surface changes of stimulated cells. Control cells (not stimulated) were smooth or provided with varying numbers of microvilli (Fig. 1). The latter, usually, were short and thin. The surface features of stimulated cells were considerably different. There was marked increase of microvilli which appeared elongated and thick. Many cells were covered with confluent polypoid projections (Fig. 2). Transmission electron microscopy demonstrated marked activity of cytoplasmic organelles. Mitochondria were increased in number and size; some giant forms with numerous cristae were observed.

Effect of Temperature, Velocity Gradient and I.V. Heparin on in Vitro Blood Coagulation and Platelet Aggregation

1967 ◽  
Vol 17 (01/02) ◽  
pp. 112-119 ◽  
Author(s):  
L Dintenfass ◽  
M. C Rozenberg
Keyword(s):  
Blood Coagulation ◽  
Velocity Gradient ◽  
Elevated Temperatures ◽  
Morphological Changes ◽  
Effect Of Temperature ◽  
Clotting Time ◽  
Progressive Change ◽  
Aggregation Of Platelets ◽  
Microscopic Techniques

SummaryA study of blood coagulation was carried out by observing changes in the blood viscosity of blood coagulating in the cone-in-cone viscometer. The clots were investigated by microscopic techniques.Immediately after blood is obtained by venepuncture, viscosity of blood remains constant for a certain “latent” period. The duration of this period depends not only on the intrinsic properties of the blood sample, but also on temperature and rate of shear used during blood storage. An increase of temperature decreases the clotting time ; also, an increase in the rate of shear decreases the clotting time.It is confirmed that morphological changes take place in blood coagula as a function of the velocity gradient at which such coagulation takes place. There is a progressive change from the red clot to white thrombus as the rates of shear increase. Aggregation of platelets increases as the rate of shear increases.This pattern is maintained with changes of temperature, although aggregation of platelets appears to be increased at elevated temperatures.Intravenously added heparin affects the clotting time and the aggregation of platelets in in vitro coagulation.

Cytotoxic Effect of the Combination of Gemcitabine and Atorvastatin Loaded in Microemulsion on the HCT116 Colon Cancer Cells

2017 ◽  
Vol 9 (2) ◽  
Author(s):  
Mayson H. Alkhatib ◽  
Dalal Al-Saedi ◽  
Wadiah S. Backer
Keyword(s):  
Colon Cancer ◽  
Cancer Cells ◽  
Anticancer Drugs ◽  
Therapeutic Potential ◽  
Morphological Changes ◽  
In Vitro Study ◽  
Colon Cancer Cells ◽  
Apoptosis Detection ◽  
Hct116 Cells

The combination of anticancer drugs in nanoparticles has great potential as a promising strategy to maximize efficacies by eradicating resistant, reduce the dosage of the drug and minimize toxicities on the normal cells. Gemcitabine (GEM), a nucleoside analogue, and atorvastatin (ATV), a cholesterol lowering agent, have shown anticancer effect with some limitations. The objective of this in vitro study was to evaluate the antitumor activity of the combination therapy of GEM and ATVencapsulated in a microemulsion (ME) formulation in the HCT116 colon cancer cells. The cytotoxicity and efficacy of the formulation were assessed by the 3- (4,5dimethylthiazole-2-yl)-2,5-diphyneltetrazolium bromide (MTT) assay. The mechanism of cell death was examined by observing the morphological changes of treated cells under light microscope, identifying apoptosis by using the ApopNexin apoptosis detection kit, and viewing the morphological changes in the chromatin structure stained with 4′,6-diamidino-2-phenylindole (DAPI) under the inverted fluorescence microscope. It has been found that reducing the concentration of GEM loaded on ME (GEM-ME) from 5μM to 1.67μM by combining it with 3.33μM of ATV in a ME formulation (GEM/2ATV-ME) has preserved the strong cytotoxicity of GEM-ME against HCT116 cells. The current study proved that formulating GEM with ATV in ME has improved the therapeutic potential of GEM and ATV as anticancer drugs.

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