Gentamycin inhibition of kcl-induced contractions of myometrium isolated from non-pregnant and pregnant cows

The aim of this study was to investigate the effects of gentamycin on KCl-induced contractions of myometrium isolated from both non-pregnant and pregnant cows. Myometrial strips were isolated from non-pregnant and pregnant cows and suspended in a jacketed organ bath filled with Krebs’ solution at 37°C (pH 7.4) continuously bubbled with 95% oxygen and 5% carbon dioxide; isometric contractions were recorded using an isometric force displacement transducer. After manifestation of spontaneous contractions, KCl (60 mM) was applied to the bath and the effects of gentamycin (300 µM, 600 µM) on the amplitude (g) and frequency of KCl-induced contractions were evaluated in 10-minute intervals. Data were statistically analysed using Student’s t-test and Wilcoxon signed-rank test. Gentamycin inhibited the frequency and amplitude of KCl-induced contractions in a concentration dependent manner. At 300 µM and 600 µM, gentamycin significantly inhibited the amplitude and reduced the frequency of contractions of myometrium isolated from both pregnant and non-pregnant cows. However, an increase in the extracellular Ca+ ion concentration virtually reversed this blockade. The results of this in vitro study indicate that gentamycin inhibits KCl-induced contractions of myometrium isolated from both non-pregnant and pregnant cows.

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Effects of gentamicin on spontaneous and agonist-induced in vitro contractions of isolated myometrial tissue from pregnant cows

Medycyna Weterynaryjna ◽

10.21521/mw.6333 ◽

2019 ◽

Vol 75 (11) ◽

pp. 6333-2019

Author(s):

MURAT YUKSEL ◽

HALIS OCAL ◽

AHMET AYAR

Keyword(s):

Contractile Activity ◽

Isometric Force ◽

Displacement Transducer ◽

Organ Bath ◽

Dependent Manner ◽

Gentamicin Sulphate ◽

Spontaneous Contractions ◽

Dose Dependent

The aim of the present study is to examine the effects of gentamicin sulphate on spontaneous, oxytocin and PGF2α induced in vitro contractions of myometrium isolated from pregnant cows. Myometrial strips were obtained from healthy pregnant cows and suspended in a covered organ bath filled with Krebs’ solution at 37°C (pH 7.4) continuously bubbled with 95% oxygen and 5% carbon dioxide: isometric contractions were recorded using an isometric force displacement transducer. After the stabilization of spontaneous contractile activity during a 90-minute equilibration period, contractions were recorded for 20 minutes (control). Gentamicin sulphate was then added to the tissue bath cumulatively and the responses were recorded every 20-minutes for each consecutive dose of gentamicin. In agonist-induced contractions, oxytocin or PGF2α was added to the tissue bath at the end of the equilibration period and the same protocol was followed to investigate the effects gentamicin on these agonist-induced contractions. Gentamicin decreased the frequency and inhibited the amplitude of the spontaneous contractions in a dose dependent manner (p < 0.05). The mean frequency and amplitude of oxytocin-induced contractions was significantly inhibited by the application of gentamicin (p < 0.05). Gentamicin also inhibited the contractions induced by PGF2α in a dose dependent manner (p < 0.05). This study showed gentamicin inhibited, depending on the dosage, oxytocin and PGF2α induced contractions of myometrium isolated from pregnant cows. Upon clinically examining the findings obtained by the study, gentamicin can be used as an antibacterial in septic abort and chorioamniotis in order to prevent premature birth, abortion and early uterus contractions. Further studies are necessary to test whether the same effect will take place in vivo and to examine the effects of longterm use of gentamicin on offsprings.

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Human Neuronal Cell Lines as An In Vitro Toxicological Tool for the Evaluation of Novel Psychoactive Substances

International Journal of Molecular Sciences ◽

10.3390/ijms22136785 ◽

2021 ◽

Vol 22 (13) ◽

pp. 6785

Author(s):

Valeria Sogos ◽

Paola Caria ◽

Clara Porcedda ◽

Rafaela Mostallino ◽

Franca Piras ◽

...

Keyword(s):

Cell Death ◽

Neuronal Cell ◽

In Vitro Study ◽

Dependent Manner ◽

Novel Psychoactive Substances ◽

Psychoactive Substances ◽

Concentration Dependent Manner ◽

Mechanisms Of Toxicity ◽

Neuronal Cell Lines

Novel psychoactive substances (NPS) are synthetic substances belonging to diverse groups, designed to mimic the effects of scheduled drugs, resulting in altered toxicity and potency. Up to now, information available on the pharmacology and toxicology of these new substances is very limited, posing a considerable challenge for prevention and treatment. The present in vitro study investigated the possible mechanisms of toxicity of two emerging NPS (i) 4′-methyl-alpha-pyrrolidinoexanophenone (3,4-MDPHP), a synthetic cathinone, and (ii) 2-chloro-4,5-methylenedioxymethamphetamine (2-Cl-4,5-MDMA), a phenethylamine. In addition, to apply our model to the class of synthetic opioids, we evaluated the toxicity of fentanyl, as a reference compound for this group of frequently abused substances. To this aim, the in vitro toxic effects of these three compounds were evaluated in dopaminergic-differentiated SH-SY5Y cells. Following 24 h of exposure, all compounds induced a loss of viability, and oxidative stress in a concentration-dependent manner. 2-Cl-4,5-MDMA activates apoptotic processes, while 3,4-MDPHP elicits cell death by necrosis. Fentanyl triggers cell death through both mechanisms. Increased expression levels of pro-apoptotic Bax and caspase 3 activity were observed following 2-Cl-4,5-MDMA and fentanyl, but not 3,4-MDPHP exposure, confirming the different modes of cell death.

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The Modulation of PCSK9 and LDLR by Supercritical CO2 Extracts of Mentha longifolia and Isolated Piperitone Oxide, an In Vitro Study

Molecules ◽

10.3390/molecules26133886 ◽

2021 ◽

Vol 26 (13) ◽

pp. 3886

Author(s):

Stefania Sut ◽

Irene Ferrarese ◽

Maria Giovanna Lupo ◽

Nicola De Zordi ◽

Elisa Tripicchio ◽

...

Keyword(s):

Mass Spectrometry ◽

Cell Lines ◽

Hepatoma Cell ◽

Density Lipoprotein ◽

In Vitro Study ◽

Volatile Constituent ◽

Dependent Manner ◽

Human Hepatoma Cell ◽

Concentration Dependent Manner

In the present study the ability of supercritical carbon dioxide (SCO2) extracts of M. longifolia L. leaves to modulate low-density lipoprotein receptor (LDLR) and proprotein convertase subtilisin/kexin type 9 (PCSK9) expression was evaluated in cultured human hepatoma cell lines Huh7 and HepG2. Two SCO2 extracts, one oil (ML-SCO2) and a semisolid (MW-SCO2), were subjected to detailed chemical characterization by mono- and bidimensional nuclear magnetic resonance (1D, 2D-NMR), gas chromatography coupled with mass spectrometry (GC-MS) and liquid chromatography coupled with mass spectrometry (LC-MS). Chemical analysis revealed significant amounts of fatty acids, phytosterols and terpenoids. ML-SCO2 was able to induce LDLR expression at a dose of 60 µg/mL in HuH7 and HepG2 cell lines. Furthermore, ML-SCO2 reduced PCSK9 secretion in a concentration-dependent manner in both cell lines. Piperitone oxide, the most abundant compound of the volatile constituent of ML-SCO2 (27% w/w), was isolated and tested for the same targets, showing a very effective reduction of PCSK9 expression. The overall results revealed the opportunity to obtain a new nutraceutical ingredient with a high amount of phytosterols and terpenoids using the SCO2 extraction of M. longifolia L., a very well-known botanical species used as food. Furthermore, for the first time we report the high activity of piperitone oxide in the reduction of PCSK9 expression.

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Thrombin activatable fibrinolysis inhibitor (TAFI) affects fibrinolysis in a plasminogen activator concentration-dependent manner

Thrombosis and Haemostasis ◽

10.1160/th03-06-0377 ◽

2004 ◽

Vol 91 (03) ◽

pp. 473-479 ◽

Cited By ~ 15

Author(s):

Ana Guimarães ◽

Dingeman Rijken

Keyword(s):

Plasminogen Activator ◽

In Vitro Study ◽

Inhibitory Effect ◽

Retardation Factor ◽

Clot Lysis ◽

Dependent Manner ◽

Plasma Clot ◽

Concentration Dependent Manner ◽

Plasmin Inhibitor

SummaryTAFIa was shown to attenuate fibrinolysis. In our in vitro study, we investigated how the inhibitory effect of TAFIa depended on the type and concentration of the plasminogen activator (PA). We measured PA-mediated lysis times of plasma clots under conditions of maximal TAFI activation by thrombin-thrombomodulin in the absence and presence of potato carboxypeptidase inhibitor. Seven different PAs were compared comprising both tPA-related (tPA, TNK-tPA, DSPA), bacterial PA-related (staphylokinase and APSAC) and urokinase-related (tcu-PA and k2tu-PA) PAs. The lysis times and the retardation factor were plotted against the PA concentration. The retardation factor plots were bell-shaped. At low PA concentrations, the retardation factor was low, probably due to the limited stability of TAFIa. At intermediate PA concentrations the retardation factor was maximal (3-6 depending on the PA), with TNK-tPA, APSAC and DSPA exhibiting the strongest effect. At high PA concentrations, the retardation factor was again low, possibly due to inactivation of TAFIa by plasmin or to a complete conversion of glu-plasminogen into lys-plasminogen. Using individual plasmas with a reduced plasmin inhibitor activity (plasmin inhibitor Enschede) the bell-shaped curve of the retardation factor shifted towards lower tPA and DSPA concentrations, but the height did not decrease. In conclusion, TAFIa delays the lysis of plasma clots mediated by all the plasminogen activators tested. This delay is dependent on the type and concentration of the plasminogen activator, but not on the fibrin specificity of the plasminogen activator. Furthermore, plasmin inhibitor does not play a significant role in the inhibition of plasma clot lysis by TAFI.

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Taurolidine Acts on Bacterial Virulence Factors and Does Not Induce Resistance in Periodontitis-Associated Bacteria—An In-Vitro Study

Antibiotics ◽

10.3390/antibiotics9040166 ◽

2020 ◽

Vol 9 (4) ◽

pp. 166

Author(s):

Sabrina Radakovic ◽

Nicola Andreoli ◽

Simon Schmid ◽

Sandor Nietzsche ◽

Jürg Zumbrunn ◽

...

Keyword(s):

Virulence Factors ◽

Bacterial Species ◽

Rare Event ◽

In Vitro Study ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Minimal Inhibitory Concentrations ◽

Development Of Resistance ◽

Subinhibitory Concentrations

The aims of the present study were: (a) to determine the mechanism of action of taurolidine against bacterial species associated with periodontal disease, and (b) to evaluate the potential development of resistance against taurolidine as compared with minocycline. After visualizing the mode of action of taurolidine by transmission electron micrographs, the interaction with most important virulence factors (lipopolysaccharide (LPS), Porphyromonas gingivalis gingipains, Aggregatibacter actinomycetemcomitans leukotoxin), was analyzed. Then, 14 clinical isolates from subgingival biofilm samples were transferred on agar plates containing subinhibitory concentrations of taurolidine or minocycline up to 50 passages. Before and after each 10 passages, minimal inhibitory concentrations (MICs) were determined. Increasing MICs were screened for efflux mechanism. Taurolidine inhibited in a concentration-dependent manner the activities of LPS and of the arginine-specific gingipains; however, an effect on A. actinomycetemcomitans leukotoxin was not detected. One P. gingivalis strain developed a resistance against taurolidine, which was probably linked with efflux mechanisms. An increase of MIC values of minocycline occurred in five of the 14 included strains after exposure to subinhibitory concentrations of the antibiotic. The present results indicate that: (a) taurolidine interacts with LPS and gingipains, and (b) development of resistance seems to be a rare event when using taurolidine.

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Enamel Surface Roughness after Lingual Bracket Debonding: An In Vitro Study

Materials ◽

10.3390/ma12244196 ◽

2019 ◽

Vol 12 (24) ◽

pp. 4196

Author(s):

Martina Eichenberger ◽

Anna Iliadi ◽

Despina Koletsi ◽

George Eliades ◽

Carlalberta Verna ◽

...

Keyword(s):

Surface Roughness ◽

In Vitro Study ◽

Rank Test ◽

Roughness Parameters ◽

Signed Rank ◽

Fine Diamond ◽

Before And After ◽

Signed Rank Test ◽

Lingual Brackets

The aim of the present study was to quantitatively assess changes in enamel roughness parameters before and after lingual bracket debonding. The lingual surface of 25 sound premolars extracted for orthodontic reasons was studied by 3D optical interferometric profilometry before and after debonding of lingual brackets following enamel finishing (with fine diamond) and polishing (with 12- and 20-fluted carbide burs). The roughness parameters tested were the amplitude parameters Sa and Sz, the hybrid parameter Sdr, and the functional parameters Sc and Sv. The parameter differences (after debonding-reference) were calculated, and statistical analysis was performed via a Wilcoxon signed-rank test. Statistically significantly higher values were observed in all the surface roughness parameters of enamel surfaces after finishing and polishing, with the mostly affected parameter being the Sdr. Under the conditions of the present study, the finishing and polishing instruments used after debonding of lingual noncustomized brackets created a surface texture rougher than the control in all the tested roughness parameters.

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Toxicity Effects of Methylene Blue on Rat Intervertebral Disc Annulus Fibrosus Cells

January 2018 - Pain Physician ◽

10.36076/ppj/2019.22.155 ◽

2019 ◽

Vol 2 (22.2) ◽

pp. 155-164

Author(s):

Liang Zhang

Keyword(s):

Methylene Blue ◽

Cell Apoptosis ◽

Annulus Fibrosus ◽

Caspase 3 ◽

In Vitro Study ◽

Collagen Type I ◽

Type I ◽

Dependent Manner ◽

Concentration Dependent Manner

Background: There is an increasing local application of methylene blue (MB) in the treatment of discogenic low back pain (LBP) and percutaneous transforaminal endoscopic discectomy (PTED) procedures. MB could generate DNA damage and induce apoptosis in different cell types; however, the effects of MB on intervertebral disc (IVD) annulus fibrosus (AF) cells are not clearly understood. Objective: The objective of this study was to investigate the effects of different concentrations of MB on rat AF cells in vitro. Study Design: This study used an experimental design. Setting: This research was conducted at the Orthopaedic Institute of the Clinical Medical College of Yangzhou University. Methods: AF cells were isolated and cultured with different concentrations of MB (0, 2, 20, and 200 μg/mL) and assessed to determine the possible cytotoxic effects of MB. The cell proliferation was detected by Cell Counting Kit-8 (CCK-8) assay. The inverted phase-contrast microscopy was used to perform morphological observation of apoptotic cells, and flow cytometry was used to measure the incidence of cell apoptosis. The mRNA and protein expression levels of apoptosis-associated genes (caspase-3, Bcl-2, and Bax) and other related genes (collagen type I, transforming growth factor β1 [TGF-β1], fibroblast growth factor [bFGF], and tissue inhibitor of metalloproteinase-1 [TIMP-1]) were analyzed by quantitative real-time PCR (RT-PCR) and Western blotting. Results: Our results indicated that MB reduced cell viability in a concentration- and timedependent manner. MB also induced marked AF cell apoptosis in a concentration-dependent manner observed by inverted phase-contrast microscopy, flow cytometry, and indicated by the increased expression of caspase-3. Both RT-PCR and Western blotting revealed significant upregulation of Bax and caspase-3 expression levels accompanied by decreased expression of Bcl2 in a concentration-dependent manner. Moreover, collagen type I, TGF-β1, bFGF, and TIMP-1 mRNA and protein levels were also found to be decreased by MB in a concentration-dependent manner. Limitations: Limitations of this study were the in vitro study design and lack of in vivo validation of the observed effects of MB on human IVD cells. Conclusions: Our results indicate that a high concentration of MB can not only inhibit proliferation and paracrine function of AF cells, but can also induce cell apoptosis in a concentration-dependent manner, suggesting that it is necessary to choose low concentrations of MB in practical application and limit the use of MB in the treatment of discogenic LBP to research protocols. Key words: Methylene blue, annulus fibrosus cell, proliferation, apoptosis, paracrine

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Influence of Thymoquinone Exposure on the Micro-Hardness of Dental Enamel: An In Vitro Study

European Journal of Dentistry ◽

10.1055/s-0039-1697117 ◽

2019 ◽

Vol 13 (03) ◽

pp. 318-322 ◽

Cited By ~ 3

Author(s):

Imran Farooq ◽

Saqib Ali ◽

Intisar Ahmad Siddiqui ◽

Khalifa S. Al-Khalifa ◽

Mohammed Al-Hariri

Keyword(s):

Artificial Saliva ◽

Dental Enamel ◽

In Vitro Study ◽

Rank Test ◽

Control Group ◽

Micro Hardness ◽

Signed Rank Test ◽

The Mean ◽

Experimental Group

Abstract Objective The aim of this study was to assess changes in micro-hardness level of enamel after it was exposed to thymoquinone (TQ). Materials and Methods Sixteen enamel blocks were prepared and divided into two groups (each group received eight blocks, n = 8); Gp 1 (control): enamel blocks kept in 100 mL artificial saliva (AS) for 24 hours and Gp 2: enamel blocks kept in a mixture of TQ powder (1 g) and AS (100 mL) for 24 hours. Post-immersion they were subjected to simulated brushing with each sample receiving 8,000 linear strokes. For brushing, 3 mL of AS and TQ oil was used for groups 1 and 2, respectively. Enamel surfaces were analyzed for changes in values of surface micro-hardness (pre-immersion, post-immersion, and post-brushing) by obtaining Vickers hardness number (VHN). Results The present study indicated improvement in micro-hardness levels for both groups although experimental group showed more enhancement. The mean baseline VHN for control group was 498.6, 500.4 for post-immersion, and 503.5 for post-brushing. The mean baseline VHN for experimental group was 448.7, 531 for post-immersion, and 610.3 for post-brushing. Statistically significant differences (p < 0.05) were observed when post-brushing VHN values of both groups were compared and also within the experimental group when post-brushing values were compared with baseline values. Statistical Analysis Wilcoxon signed-rank test was applied for the evaluation of pre- and post-exposure hardness values. Level of significance was ≤0.05. Conclusion The exposure of enamel to TQ led to an improvement in its micro-hardness levels. Further studies are required to understand the mechanism of action of TQ on human tissues.

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The Influence of Laboratory Scanner Versus Intra-Oral Scanner on Determining the Implant Axis by Using Three Different Scan Abutments

Applied Sciences ◽

10.3390/app11188543 ◽

2021 ◽

Vol 11 (18) ◽

pp. 8543

Author(s):

Asaf Shely ◽

Shiri Livne ◽

Gil Ben-Izhack ◽

Michal Lokshin ◽

Shahar Har-Nes ◽

...

Keyword(s):

Statistical Difference ◽

3D Model ◽

In Vitro Study ◽

Rank Test ◽

Rotational Angle ◽

Signed Rank ◽

Signed Rank Test ◽

3D Software

Background: The purpose of this in vitro study was to compare the implant axis’ spatial position and orientation by using laboratory scanner versus intra-oral scanner with three different scan abutments. Methods: A 3D model was printed with an internal hex implant analog in the place of teeth 35#. Three standard scan abutments were used: MIS (two-piece titanium), AB (two-piece PEEK and titanium base) and ZZ (one-piece PEEK). Each scan abutment was scanned 30 times by TRIOS E3 (laboratory scanner) and 30 times by Omnicam (intra-oral scanner). For each scan, an STL (stereolithography) file was created, and the spatial characterization of each scan abutment was measured in the X, Y, Z coordinates, and rotational and longitudinal angles. The comparison between all the scans was conducted by superimposition of the STL files, using a 3D software. A t-test and Wilcoxon signed-rank test were used. (p < 0.05) Results: Only the MIS scan abutment showed no statistical difference in the X and Z axes. (p < 0.05). All other scan abutments showed a statistical difference in all axes. The rotational angle of the AB scan abutment was twice the angle of the MIS and ZZ scan abutments. Conclusions: All three scan abutments showed a rotational deviation of the implant axis between the laboratory scanner and the intra-oral scanner. The AB scan abutment showed the greatest deviation (1.04 degrees) while the other two abutments showed deviations of about half a degree in relation to the laboratory scan abutment. There is a need for further studies which will examine the influence of geometry, material, and scan abutment parts on the accuracy of the scan obtained.

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Evaluation of fluoride releasing and recharging of orthodontic adhesive resins: An in vitro study.

Erbil Dental Journal ◽

10.15218/edj.2020.18 ◽

2020 ◽

Vol 3 (2) ◽

pp. 126-134

Author(s):

Sava Bakhtyar ◽

Bayan Hassan

Keyword(s):

Color Change ◽

In Vitro Study ◽

Rank Test ◽

Common Side Effect ◽

Control Group ◽

Fluoride Release ◽

Adhesive Resin ◽

Fixed Appliance ◽

Signed Rank Test

Background and objective: Development of white spot lesions to be a well-recognized and common side effect of orthodontic fixed appliance, in spite of vast improvement in preventive dental techniques and procedures. The aim of this study is to determine and compare amount of fluoride releasing and recharging of orthodontic adhesive resins. Materials and methods: 12 specimens for fluoride releasing and recharging were used for each tested materials, materials tested for fluoride releasing were Transbond plus color change and Resilience (fluoridated ortho-adhesive resin). Fluoride releasing was estimated daily for 14 days, then weekly up to 28 days, then at day 42, 70 days. For fluoride recharging in addition to above materials Transbond XT (non- fluoridated ortho-adhesive resin) were used as a control group. The 12 specimens of each material were divided into two equal subgroups to simulate two different methods of fluoride application, namely, brushing with fluoride-containing toothpaste and after the topical application of fluoride gel. Fluoride releasing was measured at days 1,2,3,7 for gel application while for brushing group releasing of fluoride was measured at 7 days. Results: Mann Whitney test, Kruskal Wallis test and Wilcoxon signed rank test were used for statistical analysis. During tested period all materials showed statically different capacity to release and uptake fluoride. Transbond plus color change was highest ability for fluoride releasing and recharging when compared to others materials. Conclusion: Transbond plus color change and Resilience, showed fluoride release in distal de-ionized water. For fluoride recharging Transbond plus color change had more ability for re-charging than two other materials. Keywords: Fluoride release, fluoride recharge, ortho-adhesive resin.

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