Evaluation of the effects of Rumex obtusifolius seed and leaf extracts against Acanthamoeba: An in vitro study

2020 ◽  
Vol 20 ◽  
Author(s):  
Tooran Nayeri ◽  
Farahnaz Bineshian ◽  
Fariba Khoshzaban ◽  
Abdolhossein Dalimi Asl ◽  
Fatemeh Ghaffarifar
Keyword(s):  
Flow Cytometry ◽  
Culture Medium ◽  
In Vitro Study ◽  
Vital Staining ◽  
Seed Extract ◽  
Clinical Strain ◽  
Inhibitory Effects ◽  
Leaf Extracts ◽  
Rumex Obtusifolius

Background: Acanthamoebiasis treatment is a major and challenging problem due to the presence of resistant cyst form. Many herbal extracts and their derivatives have been used against trophozoites and cysts of Acanthamoeba, but no effective therapeutic agent has yet been discovered. Therefore, the present study aimed to evaluate the effect of Rumex obtusifolius (R. obtusifolius) extracts against a clinical strain of Acanthamoeba genotype T4 in vitro. Methods: In this experimental study, after genotyping the clinical isolate, the hydroalcohlic extracts of R. obtusifolius seeds and leaves were prepared. Different concentrations (1.25, 2.5, 5 and 10 mg/ml) of extracts were tested in triplicate (24, 48 and 72h) on trophozoites and cysts of Acanthamoeba. Mortality of the parasite was assessed by trypan blue vital staining and flow cytometry analysis. Results: Results showed that the extract of R. obtusifolius leaves at the concentration of 10 mg/ml killed 100% of trophozoites and cysts after 72 h. However, the seed extract of R. obtusifolius had weak inhibitory effects on trophozoites and cysts of Acanthamoeba. In the presence of 10 mg/ml of hydroalcohlic seed extract of R.obtusifolius in culture medium after 72 h, 28.6% of trophozoites and 0% of cysts of Acanthamoeba were killed. After analysis by flow cytometry, seeds and leaves extracts indicated apoptosis effect. Seed and leaf of extracts caused 2.6% and 0.4% percent apoptosis. Conclusion: These extracts are not promising candidates for further medicine development on acanthamoebiasis. Nonetheless, further researches are necessary to clarify effective fractions of seed and leaf extracts of R. obtusifolius and their mechanisms of action.

Identifying Phenotypically Distinct Fibroblast Subsets in Type 2 Diabetes–Associated Iatrogenic Laryngotracheal Stenosis

2021 ◽  
pp. 019459982110147
Author(s):  
Ioan A. Lina ◽  
Alexandra Berges ◽  
Rafael Ospino ◽  
Ruth J. Davis ◽  
Kevin M. Motz ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Flow Cytometry ◽  
Pearson Correlation ◽  
Phenotypic Expression ◽  
In Vitro Study ◽  
Cell Protein ◽  
Laryngotracheal Stenosis ◽  
Tertiary Referral Center

Objective Iatrogenic laryngotracheal stenosis (iLTS) is the pathologic narrowing of the glottis, subglottis, and/or trachea secondary to intubation or tracheostomy related injury. Patients with type 2 diabetes mellitus (T2DM) are more likely to develop iLTS. To date, the metabolomics and phenotypic expression of cell markers in fibroblasts derived from patients with T2DM and iLTS are largely unknown. Study Design Controlled in vitro cohort study. Setting Tertiary referral center (2017-2020). Methods This in vitro study assessed samples from 6 patients with iLTS who underwent surgery at a single institution. Fibroblasts were isolated from biopsy specimens of laryngotracheal scar and normal-appearing trachea and compared with controls obtained from the trachea of rapid autopsy specimens. Patients with iLTS were subcategorized into those with and without T2DM. Metabolic substrates were identified by mass spectrometry, and cell protein expression was measured by flow cytometry. Results T2DM iLTS-scar fibroblasts had a metabolically distinct profile and clustered tightly on a Pearson correlation heat map as compared with non-T2DM iLTS-scar fibroblasts. Levels of itaconate were elevated in T2DM iLTS-scar fibroblasts. Flow cytometry demonstrated that T2DM iLTS-scar fibroblasts were associated with higher CD90 expression (Thy-1; mean, 95%) when compared with non-T2DM iLTS-scar (mean, 83.6%; P = .0109) or normal tracheal fibroblasts (mean, 81.1%; P = .0042). Conclusions Scar-derived fibroblasts from patients with T2DM and iLTS have a metabolically distinct profile. These fibroblasts are characterized by an increase in itaconate, a metabolite related to immune-induced scar remodeling, and can be identified by elevated expression of CD90 (Thy-1) in vitro.

scholarly journals Cytokinin Mediated Increased In Vitro Production of Secondary Metabolites with Special Reference to Solasodine in Solanum erianthum

2021 ◽  
Vol 8 (02) ◽  
pp. e62-e68
Author(s):  
Jeeta Sarkar ◽  
Nirmalya Banerjee
Keyword(s):  
Secondary Metabolites ◽  
High Performance ◽  
Culture Medium ◽  
In Vitro Production ◽  
Leaf Extracts ◽  
Plant Parts ◽  
Regenerated Plants ◽  
Vitro Production ◽  
Regenerated Plantlets

AbstractSteroid alkaloid solasodine is a nitrogen analogue of diosgenin and has great importance in the production of steroidal medicines. Solanum erianthum D. Don (Solanaceae) is a good source of solasodine. The aim of this study was to evaluate the effect of different cytokinins on the production of secondary metabolites, especially solasodine in the in vitro culture of S. erianthum. For solasodine estimation, field-grown plant parts and in vitro tissues were extracted thrice and subjected to high-performance liquid Chromatography. Quantitative analysis of different secondary metabolites showed that the amount was higher in the in vitro regenerated plantlets compared to callus and field-grown plants. The present study critically evaluates the effect of the type of cytokinin used in the culture medium on solasodine accumulation in regenerated plants. The highest solasodine content (46.78±3.23 mg g-1) was recorded in leaf extracts of the in vitro grown plantlets in the presence of 6-γ,γ-dimethylallylamino purine in the culture medium and the content was 3.8-fold higher compared to the mother plant.

scholarly journals Antibacterial Inhibitory Effects of Punica Granatum Gel on Cariogenic Bacteria: An in vitro Study

2017 ◽  
Vol 10 (2) ◽  
pp. 152-157 ◽  
Author(s):  
Grazielle Millo ◽  
Apa Juntavee ◽  
Ariya Ratanathongkam ◽  
Natsajee Nualkaew ◽  
Peerapattana, Jomjai ◽  
...  
Keyword(s):  
High Performance ◽  
In Vitro Study ◽  
Punica Granatum ◽  
Antibacterial Activities ◽  
Diffusion Method ◽  
Inhibitory Effects ◽  
Cariogenic Bacteria ◽  
Zones Of Inhibition ◽  
Time Kill

ABSTRACT Aim This study evaluated the in vitro antibacterial effects of the formulated Punica granatum (PG) gel against Streptococcus mutans, Streptococcus sanguinis, and Lactobacillus casei. Materials and methods The PG extract was dissolved in water at 500 mg/mL. High performance liquid chromatography (HPLC) was used for identification and quantification of chemical marker punicalagin. Minimum bactericidal concentration (MBC) and time-kill assay (TKA) were investigated. Antibacterial activities of the formulated PG gel, 2% chlorhexidine (CHX) gel and blank gel were tested by measuring the zones of inhibition through agar well diffusion method. Results The HPLC results showed presence of punicalagin at 2023.58 ± 25.29 μg/mL in the aqueous PG extract and at 0.234% (w/w) in the formulated PG gel. The MBC for S. mutans, S. Sanguinis, and L. casei were 250, 125, and 500 mg/mL respectively. The TKA of 500 mg/mL aqueous PG extract showed total inhibition of S. mutans, S. Sanguinis, and L. casei at 6, 1, and 24 hours contact time respectively. Agar well diffusion revealed that for S. mutans, CHX gel > PG gel > blank gel; for S. sanguinis, CHX gel = PG gel > blank gel; for L. casei, CHX gel > PG gel = blank gel. Comparison of the PG gel potency showed that S. sanguinis = S. mutans > L. casei. Conclusion The PG gel equivalent to 0.234% punicalagin (w/w) inhibited S. mutans and S. sanguinis but not L. casei within 24 hours incubation period and has the potential to be used for caries prevention. How to cite this article Millo G, Juntavee A, Ratanathongkam A, Nualkaew N, Peerapattana J, Chatchiwiwattana S. Antibacterial Inhibitory Effects of Punica Granatum Gel on Cariogenic Bacteria: An in vitro Study. Int J Clin Pediatr Dent 2017;10(2):152-157.

scholarly journals ASSESSMENT OF GENETIC STABILITY OF MICROPROPAGATED Eucalyptus globulus Labill HYBRID CLONES BY MEANS OF FLOW CYTOMETRY AND MICROSATELLITES MARKERS

2017 ◽  
Vol 41 (1) ◽  
Author(s):  
Leandro Silva Oliveira ◽  
Aloisio Xavier ◽  
Wagner Campos Otoni ◽  
José Marcello Salabert Campos ◽  
Lyderson Facio Viccini ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Microsatellite Markers ◽  
Genetic Stability ◽  
Culture Medium ◽  
Eucalyptus Grandis ◽  
Genetic Fidelity ◽  
Shoot Multiplication ◽  
Micropropagated Plants ◽  
Hybrid Clones

ABSTRACT Flow cytometry and microsatellite markers were used to determine a genetic fidelity of micropropagated plants from the two Eucalyptus urophylla x E. globulus clones and a Eucalyptus grandis x E. globulus clone derived from adult material. Clones were repeatedly subcultured for 25 subcultures on MS medium supplemented with BA (2.22 µM) and ANA (0.05 µM) for in vitro shoot multiplication. The elongation was performed in MS culture medium supplemented with AIB (2.46 µM) and BA(0.22 µM). The ex vitro rooting and acclimatization phases were lead at the same time. The micropropagated clones showed genetic stability by flow cytometry and microsatellite markers. The results proved that micropropagation, for purposes of rejuvenation, can be a viable technique to generate genetically stable or identical E. globulus hybrid clones.

scholarly journals Effect of Lignocaine Concentration on Human Fibroblasts Growth in Eyes Undergoing Trabeculectomy: An in vitro Study

2018 ◽  
Vol 3 (3) ◽  
pp. 1-10 ◽  
Author(s):  
Madhuravasal Krishnan Janani ◽  
Venkatakrishnan Jaichandran ◽  
Hajib Narahari Rao Madhavan ◽  
Lingam Vijaya ◽  
Ronnie Jacob George ◽  
...  
Keyword(s):  
Culture Medium ◽  
In Vitro Model ◽  
Morphological Changes ◽  
Human Fibroblasts ◽  
Annexin V ◽  
In Vitro Study ◽  
Tenon’S Capsule ◽  
Tenon's Capsule ◽  
Vitro Model

Purpose: To evaluate the effect of lignocaine on growth and apoptosis indication of primary human Tenon’s capsule fibroblast (HTFs) in an in vitro model. Patients and Methods: Tenon’s capsule tissue obtained from patients undergoing trabeculectomy were grown in cell culture medium. The effect of different concentrations of lignocaine (0.5, 1.0, 1.5, and 2%) on the morphology and growth of the fibroblasts was studied using microscopy, cell viability, and proliferation assay, and apoptosis was detected using the FITC Annexin V Apoptosis Kit. Results: Morphological changes similar to those of apoptotic cells, including cytoplasmic vacuolation, shrinkage, and rounding were visualized in the cells treated with concentrations greater than 1.0% (i.e., 1.5, 2.0%). Though proliferation inhibition was found with all four concentrations (0.5–2.0%), the viability of cells decreased from 1.0% lignocaine. Conclusion: 0.5% lignocaine prevents proliferation of fibroblasts without causing apoptosis in vitro.

Refining insulin concentrations in culture medium containing growth factors BMP15 and GDF9: An in vitro study of the effects on follicle development of goats

2017 ◽  
Vol 185 ◽  
pp. 118-127 ◽  
Author(s):  
D.J. Dipaz-Berrocal ◽  
N.A.R. Sá ◽  
D.D. Guerreiro ◽  
J.J.H. Celestino ◽  
J. Leiva-Revilla ◽  
...  
Keyword(s):  
Growth Factors ◽  
Culture Medium ◽  
In Vitro Study ◽  
Vitro Study ◽  
Follicle Development

scholarly journals Ascorbic acid attenuates activation and cytokine production in sepsis-like monocytes

2021 ◽  
Author(s):  
Tobias Schmidt ◽  
Robin Kahn ◽  
Fredrik Kahn
Keyword(s):  
Flow Cytometry ◽  
Ascorbic Acid ◽  
Cytokine Production ◽  
In Vitro Study ◽  
Surface Expression ◽  
High Dose ◽  
Functional Assay ◽  
Dependent Manner ◽  
Dose Dependent

Objective To investigate the effects of high dose ascorbic acid (AA) on monocyte polarization and cytokine production in vitro Design Experimental in vitro study of cells from healthy subjects and patients with sepsis Setting University research laboratory and academic hospital Subjects Six healthy controls and three patients with sepsis Interventions Monocytes were isolated from whole blood of healthy donors (n=6) and polarized in vitro for 48hrs using LPS or LTA. Polarization was confirmed by surface marker expression using flow cytometry. As a comparison, monocytes were also isolated from septic patients (n=3) and analyzed for polarization markers. The effect of AA on monocyte polarization was evaluated. As a functional assay, AA-treated monocytes were analyzed for cytokine production of TNF and IL-8 by intracellular staining and flow cytometry following activation with LPS or LTA. Measurements and Main Results Both LPS and LTA induced polarization in healthy monocytes in vitro, with increased expression of both pro- (CD40 and PDL1, p<0.05) and anti-inflammatory (CD16 and CD163, p<0.05) polarization markers, with non-significant effects on CD86 and CD206. This pattern resembled, at least partly, that of monocytes from septic patients. Treatment with AA significantly inhibited the upregulation of surface expression of CD16 and CD163 (p<0.05) in a dose dependent manner, but not CD40 or PDL-1. Finally, AA attenuated LPS or LTA-induced cytokine production of IL-8 and TNF in a dose-dependent manner (both p<0.05). Conclusions AA inhibits upregulation of anti-, but not pro-inflammatory related markers in LPS or LTA polarized monocytes. Additionally, AA attenuates cytokine production from in vitro polarized monocytes, displaying functional involvement. This study provides important insight into the immunological effects of high dose AA on monocytes, and potential implications in sepsis.

Antilisterial Effects of a Sorbate-Nisin Combination In Vitro and on Packaged Beef at Refrigeration Temperature

1997 ◽  
Vol 60 (9) ◽  
pp. 1075-1080 ◽  
Author(s):  
SHERYL M. AVERY ◽  
SAVA BUNCIC
Keyword(s):  
Carbon Dioxide ◽  
In Vitro Study ◽  
Clinical Strain ◽  
Lag Phase ◽  
Initial Population ◽  
Pathogen Load ◽  
Beef Steaks ◽  
Packaged Beef ◽  
Phosphate Buffered Saline

The antilisterial effects of a sorbate-nisin combination were assessed in vitro and on beef at refrigeration temperature. Three hemolytic pathogenic strains of Listeria monocytogenes, reference strain NCTC 7973, food strain L70, and clinical strain L94, were stored at 4°C in phosphate-buffered saline, pH 5.5, containing a combination of sorbate (0.2% wt/vol) and nisin (40 IU/ml). After 4 weeks, hemolysin production by the strains had ceased, their subsequent lag phases at 37°C were extended from an initial 1.23 to 1.32 h to a final 7.13 to 8.06 h and their pathogenicity for chick embryos had decreased from an initial 93.3 to 95.5% to a final 43.3 to 60.0%. Sterile beef steaks of normal pH (5.4 to 5.5) were inoculated with a cocktail of the three strains at approximately 5 log CFU/cm2 and the surface of half the steaks was treated with the antimicrobial solution 1.0% sorbate plus 1,000 IU of nisin per ml. The meat was packaged under vacuum or 100% carbon dioxide and stored at 4°C for 4 weeks. On untreated meat, L. monocytogenes grew by 1.79 log cycles in vacuum packages, but in CO2 packages the initial population decreased by 0.54 log cycle. On treated vacuum-packaged meat, L. monocytogenes decreased during storage to the extent that 96.5% of the initial pathogen load was eliminated, but the lag phase of the remaining cells at 37°C was unaffected. On treated CO2-packaged meat L. monocytogenes decreased during storage to the extent that 89.3% of the initial pathogen load was eliminated, and for surviving cells the lag phase at 37°C was extended. Treatment with the sorbate-nisin combination did not significantly affect pathogenicity of the L. monocytogenes cocktail recovered from vacuum- or carbon dioxide-packages after storage, in contrast to the in vitro study, where pathogenicity was clearly attenuated. The reason for this difference is unknown.

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