Effects of Solvents, Emulsions, Cosolvents, and Complexions on Ex Vivo Mouse Myometrial Contractility

AbstractA great need exists to develop tocolytic and uterotonic drugs that combat poor, labor-related maternal and fetal outcomes. A widely utilized method to assess novel compounds for their tocolytic and uterotonic efficacy is the isometric organ bath contractility assay. Unfortunately, water-insoluble compounds can be difficult to test using the physiological, buffer-based, organ bath assay. Common methods for overcoming solubility issues include solvent variation, cosolvency, surfactant or complexion use, and emulsification. However, these options for drug delivery or formulation can impact tissue function. Therefore, the goal of this study was to evaluate the ability of common solvents, surfactants, cosolvents, and emulsions to adequately solubilize compounds in the organ bath assay without affecting mouse myometrial contractility. We found that acetone, acetonitrile, and ethanol had the least effect, while dimethylacetamide, ethyl acetate, and isopropanol displayed the greatest inhibition of myometrial contractility based on area under the contractile curve analyses. The minimum concentration of surfactants, cosolvents, and human serum albumin required to solubilize nifedipine, a current tocolytic drug, resulted in extensive bubbling in the organ bath assay, precluding their use. Finally, we report that an oil-in-water base emulsion containing no drug has no statistical effect beyond the control (water), while the drug emulsion yielded the same potency and efficacy as the freely solubilized drug.

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Myometrial Responses to Beta-Adrenoceptor Antagonists in Gynecological Malignancies

Gynecologic and Obstetric Investigation ◽

10.1159/000513718 ◽

2021 ◽

pp. 1-8

Author(s):

Beata Modzelewska ◽

Marcin Jóźwik ◽

Tomasz Kleszczewski ◽

Stanisław Sulkowski ◽

Maciej Jóźwik

Keyword(s):

Ovarian Cancer ◽

Cervical Cancer ◽

Reference Group ◽

Ex Vivo ◽

Contractile Activity ◽

Organ Bath ◽

Human Uterus ◽

Uterine Contractility ◽

Beta Adrenoceptor ◽

Gynecological Malignancies

Objective: The aim of the study was to determine the influence of beta-adrenoceptor (ADRB) antagonists on contractile activity of the nonpregnant human uterus in patients affected by gynecological malignancies. Design: This was a controlled and prospective ex vivo study. Setting: The work was conducted as a collaboration between 4 academic departments. Materials and Methods: Myometrial specimens were obtained from women undergoing hysterectomy for benign gynecological disorders (reference group; N = 15), and ovarian (N = 15), endometrial (N = 15), synchronous ovarian-endometrial (N = 3), and cervical cancer (N = 10). Contractions of myometrial strips in an organ bath before and after applications of ADRB antagonists (propranolol, bupranolol, SR 59230A, and butoxamine) were studied under isometric conditions. Results: Propranolol and bupranolol attenuated contractions in the endometrial and cervical cancer groups similar to that in the reference group (all p < 0.05), whereas opposite effects were observed in the ovarian and synchronous ovarian-endometrial cancer groups. SR 59230A and butoxamine significantly increased contractions in the ovarian cancer group (both p < 0.001). Limitations: These results require now to be placed into a firm clinical context. Conclusions: Our study indicates that ovarian cancer considerably alters contractile activity of the nonpregnant human uterus in response to ADRB antagonists. This suggests a pathogenetic role of beta-adrenergic pathways in this malignancy. Furthermore, propranolol and bupranolol substantially influence spontaneous uterine contractility.

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Compared Phenolic Compound Contents of 22 Commercial Fruit and Vegetable Juices: Relationship to Ex-Vivo Vascular Reactivity and Potential In Vivo Projection

Antioxidants ◽

10.3390/antiox9020092 ◽

2020 ◽

Vol 9 (2) ◽

pp. 92 ◽

Cited By ~ 1

Author(s):

Alexis Matute ◽

Jessica Tabart ◽

Jean-Paul Cheramy-Bien ◽

Bernard Pirotte ◽

Claire Kevers ◽

...

Keyword(s):

Phenolic Compounds ◽

Phenolic Compound ◽

Vascular Reactivity ◽

Ex Vivo ◽

Clinical Investigation ◽

Epigallocatechin Gallate ◽

Fruit Juices ◽

Organ Bath ◽

Total Polyphenol ◽

Total Anthocyanin

The real impact of polyphenol-rich vegetable and fruit juice intake on cardiovascular health remains a matter of controversy. In the present study, rat aorta segments immersed in an organ bath (OB) were used to explore whether the total polyphenol content and/or individual phenolic compound contents of 22 commercial vegetable (n = 3) and fruit juices [(citrus (n = 5), berries (n = 10), apple (n = 2), pineapple (n = 2)] might be associated with vascular tone. Red juices (particularly blackcurrant) and lemon juice caused the most marked vasorelaxation, its amplitude being endothelium dependent or not according to the volume ratio of juice to initial OB solution Vjuice/VOBS). At volume ratios 5% and 10%, both the juice and OB total polyphenol for all juices and total anthocyanin contents for berry juices significantly correlated with aorta vasorelaxation intensity. This was not the case for total or individual flavonols (except kaempferol) or for total or individual flavanols (except epigallocatechin gallate). If one relates our measured concentrations of individual phenolic compounds in OB to what is known about their physiological concentrations, and given our evidenced correlations between compound concentrations and vasorelaxation intensity, kaempferol, epigallocatechin gallate and peonidin-3-O-glucoside seem to emerge as the interesting phenolic compounds likely to be responsible for the potent vasorelaxation observed with fruit juices, and more particularly blackcurrant ones. Clinical investigation is required, however, to confirm our observations.

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Glucose uptake during contraction in isolated skeletal muscles from neuronal nitric oxide synthase μ knockout mice

Journal of Applied Physiology ◽

10.1152/japplphysiol.00056.2015 ◽

2015 ◽

Vol 118 (9) ◽

pp. 1113-1121 ◽

Cited By ~ 12

Author(s):

Yet Hoi Hong ◽

Tony Frugier ◽

Xinmei Zhang ◽

Robyn M. Murphy ◽

Gordon S. Lynch ◽

...

Keyword(s):

Nitric Oxide ◽

Skeletal Muscle ◽

Nitric Oxide Synthase ◽

Glucose Uptake ◽

Ex Vivo ◽

Organ Bath ◽

Skeletal Muscle Glucose Uptake ◽

Nos Inhibitor ◽

Amp Activated Protein Kinase ◽

Oxide Synthase

Inhibition of nitric oxide synthase (NOS) significantly attenuates the increase in skeletal muscle glucose uptake during contraction/exercise, and a greater attenuation is observed in individuals with Type 2 diabetes compared with healthy individuals. Therefore, NO appears to play an important role in mediating muscle glucose uptake during contraction. In this study, we investigated the involvement of neuronal NOSμ (nNOSμ), the main NOS isoform activated during contraction, on skeletal muscle glucose uptake during ex vivo contraction. Extensor digitorum longus muscles were isolated from nNOSμ−/−and nNOSμ+/+mice. Muscles were contracted ex vivo in a temperature-controlled (30°C) organ bath with or without the presence of the NOS inhibitor NG-monomethyl-l-arginine (L-NMMA) and the NOS substrate L-arginine. Glucose uptake was determined by radioactive tracers. Skeletal muscle glucose uptake increased approximately fourfold during contraction in muscles from both nNOSμ−/−and nNOSμ+/+mice. L-NMMA significantly attenuated the increase in muscle glucose uptake during contraction in both genotypes. This attenuation was reversed by L-arginine, suggesting that L-NMMA attenuated the increase in muscle glucose uptake during contraction by inhibiting NOS and not via a nonspecific effect of the inhibitor. Low levels of NOS activity (∼4%) were detected in muscles from nNOSμ−/−mice, and there was no evidence of compensation from other NOS isoform or AMP-activated protein kinase which is also involved in mediating muscle glucose uptake during contraction. These results indicate that NO regulates skeletal muscle glucose uptake during ex vivo contraction independently of nNOSμ.

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PENENTUAN STABILITAS SEDIAAN KRIM TABIR SURYA DARI BAHAN EKSTRAK RIMPANG KENCUR (Kaempferia galanga L.)

Journal of Biological Researches ◽

10.23869/bphjbr.10.2.20055 ◽

2005 ◽

Vol 10 (2) ◽

pp. 103-105

Author(s):

Widji Soeratri ◽

Noor Ifansyah ◽

Diana Fitrianingrum

Keyword(s):

Chemical Stability ◽

Room Temperature ◽

Percolation Method ◽

Kaempferia Galanga ◽

Oil In Water ◽

Water Base

The objective of this research was to know the chemical stability of the etil para-metoksi sinamat (EPMS) formulated in sunscreen cream of oil in water base. The EPMS was isolated from Kaempferia galanga L. tuber pulvis with maceration-percolation method using ethanol 96 percent. The chemical stability of the EPMS formulated in sunscreen cream of the oil in water base was investigated by keeping the cream at room temperature. At appropriate interval the concentration of EPMS in cream was measured. The results showed that the EPMS formulated in sunscreen cream of oil in water base was unstable at room temperature for 70 days. The concentration of EPMS in cream significantly decreased after 70 days (p = 0,01) during keeping at room temperature with the coefficient degradation value (k)= 4.4.10-3/day.

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Rhythmic changes in colonic motility are regulated by period genes

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00402.2009 ◽

2010 ◽

Vol 298 (2) ◽

pp. G143-G150 ◽

Cited By ~ 47

Author(s):

Willemijntje A. Hoogerwerf ◽

Vahakn B. Shahinian ◽

Germaine Cornélissen ◽

Franz Halberg ◽

Jonathon Bostwick ◽

...

Keyword(s):

Knockout Mice ◽

Clock Genes ◽

Ex Vivo ◽

Colonic Motility ◽

Biological Clock ◽

Circular Muscle ◽

Organ Bath ◽

Wild Type ◽

Double Knockout ◽

Muscle Contractility

Human bowel movements usually occur during the day and seldom during the night, suggesting a role for a biological clock in the regulation of colonic motility. Research has unveiled molecular and physiological mechanisms for biological clock function in the brain; less is known about peripheral rhythmicity. This study aimed to determine whether clock genes such as period 1 ( per1) and period2 ( per2) modulate rhythmic changes in colonic motility. Organ bath studies, intracolonic pressure measurements, and stool studies were used to examine measures of colonic motility in wild-type and per1per2 double-knockout mice. To further examine the mechanism underlying rhythmic changes in circular muscle contractility, additional studies were completed in neuronal nitric oxide synthase (nNOS) knockout mice. Intracolonic pressure changes and stool output in vivo, and colonic circular muscle contractility ex vivo, are rhythmic with greatest activity at the start of night in nocturnal wild-type mice. In contrast, rhythmicity in these measures was absent in per1per2 double-knockout mice. Rhythmicity was also abolished in colonic circular muscle contractility of wild-type mice in the presence of Nω-nitro-l-arginine methyl ester and in nNOS knockout mice. These findings suggest that rhythms in colonic motility are regulated by both clock genes and a nNOS-mediated inhibitory process and suggest a connection between these two mechanisms.

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N-Acetylcysteine Does Not Improve the Endothelial and Smooth Muscle Function in the Human Saphenous Vein

Vascular and Endovascular Surgery ◽

10.1177/1538574407299618 ◽

2007 ◽

Vol 41 (3) ◽

pp. 239-245 ◽

Cited By ~ 7

Author(s):

Muhammad Anees Sharif ◽

Ulvi Bayraktutan ◽

Ian Stuart Young ◽

Chee Voon Soong

Keyword(s):

Smooth Muscle ◽

Saphenous Vein ◽

Muscle Function ◽

Ex Vivo ◽

Smooth Muscle Relaxation ◽

Organ Bath ◽

Human Saphenous Vein ◽

Vein Segment ◽

Significant Difference ◽

Smooth Muscle Function

Oxidative stress can lead to vein graft dysfunction in the saphenous vein. This ex vivo study is aimed to compare the effects of increasing concentrations of the antioxidant N-acetylcysteine (NAC) with heparinized saline (HS) on endothelial and smooth muscle function in the human saphenous vein. Long saphenous vein segment obtained during infrainguinal bypass surgery was divided into 7 rings; 1 immersed in HS and the remaining 6 in increasing NAC concentrations (0.0025%, 0.005%, 0.01%, 0.02%, 0.03%, and 0.04%). Rings were mounted in an organ bath, and relaxant responses to acetylcholine and sodium nitroprusside were assessed through isometric tension studies. Endothelium-dependent relaxations were observed in 77 vein segments from 11 patients. No significant difference was seen in veins treated with either lower NAC concentrations (0.0025%, 0.005%, 0.01%, 0.02%, and 0.03%) or HS. However, HS-treated veins showed significantly better relaxation compared to those treated with maximum (0.04%) NAC ( P < .05). Endothelium-independent relaxations were observed in 91 segments from 13 patients. No difference in relaxation was observed between veins treated with HS or any of the NAC concentrations. In conclusion, lower NAC concentrations do not offer better endothelial protection than HS, whereas the highest NAC concentration has a detrimental effect on endothelium-dependent relaxation. Moreover, NAC did not show beneficial effect on direct smooth muscle relaxation.

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Effects of Sevoflurane and Adenosine Receptor Antagonist on the Sugammadex-Induced Recovery from Rocuronium-Induced Neuromuscular Blockade: An Ex-vivo Study

10.21203/rs.3.rs-43525/v1 ◽

2020 ◽

Author(s):

Yong Beom Kim ◽

Jae-Moon Choi ◽

Chungon Park ◽

Hey-Ran Choi ◽

Junyong In ◽

...

Keyword(s):

Neuromuscular Blockade ◽

Adenosine Receptor ◽

Ex Vivo ◽

Neuromuscular Blocking ◽

Blocking Effect ◽

Organ Bath ◽

Sprague Dawley ◽

Buffer Solution ◽

Recovery Pattern ◽

Krebs Buffer

Abstract Background: Sevoflurane affects on the A1 receptor in the central nervous system (CNS) and potentiates the action of neuromuscular blocking agents. In the present study, we investigated whether sevoflurane (SEVO) has the ability to potentiate the neuromuscular blocking effect of rocuronium and if the specific antagonist of adenosine receptor (SLV320) can reverse this effect. Methods: Phrenic nerve–hemidiaphragm tissue specimens were obtained from forty Sprague-Dawley (SD) rats. The specimens were immersed in an organ bath filled with Krebs buffer and stimulated by a train-of-four (TOF) pattern using indirect supramaximal stimulation at 20 s intervals. The specimens were randomly allocated to control, 2-chloroadenosine (CADO), SEVO, or SLV320+SEVO groups. In the CADO and SLV320+SEVO groups, CADO and SLV320 were added to the organ bath from the start to a concentration of 10 μM and 10 nM, respectively. We then proceeded with rocuronium-induced blockade of >95% depression of the first twitch tension of TOF (T1) and TOF ratio (TOFR). In the SEVO and SLV320+SEVO groups, SEVO was added to the Krebs buffer solution to concentration of 400 - 500 μM for 10 min. Sugammadex-induced T1 and TOFR recovery was monitored for 30 min until >95% of T1 and >0.9 of TOFR were confirmed, and the recovery pattern was compared by plotting these data. Results: There were no significant differences in the recovery pattern between the control and SEVO groups. However, there were significant differences between the SEVO and SLV320+SEVO groups. Conclusion: Sevoflurane potentiates of rocuronium-induced neuromuscular blocking effect and delays sugammadex-induced recovery from neuromuscular blockade.

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Substance P is essential for maintaining gut muscle contractility: a novel role for coneurotransmission revealed by botulinum toxin

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00436.2012 ◽

2014 ◽

Vol 306 (10) ◽

pp. G839-G848 ◽

Cited By ~ 6

Author(s):

Cuiping Li ◽

Maria-Adelaide Micci ◽

Karnam S. Murthy ◽

Pankaj Jay Pasricha

Keyword(s):

Smooth Muscle ◽

Botulinum Toxin ◽

Substance P ◽

Motor Neurons ◽

Ex Vivo ◽

Organ Bath ◽

Excitatory Neurotransmitter ◽

Vesicular Release ◽

Nk1 Receptor Antagonist ◽

Pyloric Muscle

Substance P (SP) is commonly coexpressed with ACh in enteric motor neurons, and, according to the classical paradigm, both these neurotransmitters excite smooth muscle via parallel pathways. We hypothesized that, in addition, SP was responsible for maintaining the muscular responsiveness to ACh. We tested this hypothesis by using botulinum toxin (BoNT/A), a known blocker of vesicular release of neurotransmitters including ACh and neuropeptides. BoNT/A was injected into rat pyloric sphincter in different doses; as control we used boiled BoNT/A. At the desired time point, pylorus was dissected out and pyloric contractility was measured ex vivo in an organ bath and by measuring phosphorylation of myosin light chain 20 (MLC20). BoNT/A (10 IU) significantly reduced the response of pyloric muscle to exogenous ACh, an effect that was accompanied by reduced MLC20 phosphorylation in the muscle. Both effects were reversed by exogenous SP. CP-96345, a NK1 receptor antagonist, blocked the ability of exogenous SP to reverse the cholinergic hyporesponsiveness as well as the reduction in MLC20 phosphorylation induced by BoNT/A. In conclusion, we have identified a novel role for SP as a coneurotransmitter that appears to be important for the maintenance of muscular responsiveness to the principal excitatory neurotransmitter, ACh. These results also provide new insight into the effects of botulinum toxin on the enteric nervous system and gastrointestinal smooth muscle.

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Effects of nano-sizing on lipid bioaccessibility and ex vivo bioavailability from EPA-DHA rich oil in water nanoemulsion

Food Chemistry ◽

10.1016/j.foodchem.2018.09.084 ◽

2019 ◽

Vol 275 ◽

pp. 135-142 ◽

Cited By ~ 9

Author(s):

Tanmoy Kumar Dey ◽

Hemanta Koley ◽

Mahua Ghosh ◽

Sanjit Dey ◽

Pubali Dhar

Keyword(s):

Ex Vivo ◽

Oil In Water

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A novel GABAA receptor ligand MIDD0301 with limited blood-brain barrier penetration relaxes airway smooth muscle ex vivo and in vivo

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00356.2018 ◽

2019 ◽

Vol 316 (2) ◽

pp. L385-L390 ◽

Cited By ~ 5

Author(s):

Gene T. Yocum ◽

Jose F. Perez-Zoghbi ◽

Jennifer Danielsson ◽

Aisha S. Kuforiji ◽

Yi Zhang ◽

...

Keyword(s):

Smooth Muscle ◽

Airway Smooth Muscle ◽

Ex Vivo ◽

Drug Candidate ◽

Organ Bath ◽

Forced Oscillation Technique ◽

Gaba A ◽

Peripheral Airways ◽

Trace Concentrations

Airway smooth muscle (ASM) cells express GABA A receptors (GABAARs), and previous reports have demonstrated that GABAAR activators relax ASM. However, given the activity of GABAARs in central nervous system inhibitory neurotransmission, concern exists that these activators may lead to undesirable sedation. MIDD0301 is a novel imidazobenzodiazepine and positive allosteric modulator of the GABAAR with limited brain distribution, thus eliminating the potential for sedation. Here, we demonstrate that MIDD0301 relaxes histamine-contracted guinea pig ( P < 0.05, n = 6–9) and human ( P < 0.05, n = 6–10) tracheal smooth muscle ex vivo in organ bath experiments, dilates mouse peripheral airways ex vivo in precision-cut lung-slice experiments ( P < 0.001, n = 16 airways from three mice), and alleviates bronchoconstriction in vivo in mice, as assessed by the forced-oscillation technique ( P < 0.05, n = 6 mice). Only trace concentrations of the compound were detected in the brains of mice after inhalation of nebulized 5 mM MIDD0301. Given its favorable pharmacokinetic properties and demonstrated ability to relax ASM in a number of clinically relevant experimental paradigms, MIDD0301 is a promising drug candidate for bronchoconstrictive diseases, such as asthma.

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