Aqueous Dimethyl Sulfoxide Solutions Research Articles

A new two-dimensional, spiropyran-based polymer fluorescent nanoprobe with quantitative-fluorescent and photochromic properties for multi-substance detection

One challenge of the structural design of a fluorescent probe is how to improve the detection performance on trace target analytes in complex samples. Herein a new polymer fluorescent nanoprobe (2DSP-C28) has been synthesized, by adopting a two-dimensional (2D), spiropyran (SP)-based nanosheet structure with hydrophobic long-chain alkanes (C28). Unlike a traditional SP-based small molecule probe, the 2DSP-C28 probe can exhibit quantitative-fluorescent and photochromic properties. Under the detection of metal-ions, the nanoprobe in dimethyl sulfoxide aqueous solution is selectively fluorescent-quenched-responsive for Fe-ions (∼100 μM), with a characteristic stoichiometric ratio of <10, a high sensitivity (limit of detection: ∼0.2 μM). When the nanoprobe is incorporated into electrospun polyethylene oxide, it can be used for gas detection, and display a color-change with acid-base gas and identify the HF gas. It is expected that this new polymer fluorescent nanoprobe can be promisingly applied for rapidly environmental monitoring on the ion or gas pollution.

Towards liquid EPR dosimetry using nitroxides in aqueous solution.

Objective. Water-equivalent dosimeters are desirable for dosimetry in radiotherapy. The present work investigates basic characteristics of novel aqueous detector materials and presents a signal loss approach for electron paramagnetic resonance (EPR) dosimetry.Approach. The proposed principle is based on the radiation dose dependent annihilation of EPR active nitroxides (NO·) in aqueous solutions. Stable nitroxide radicals (3-Maleimido-2,2,5,5-tetramethyl-1-pyrrolidinyloxy (MmP), 3-Carbamoyl-2,2,5,5-tetramethyl-1-pyrrolidinyloxy (CmP)) in aqueous solutions containing dimethyl sulfoxide (DMSO) as an additive were filled in glass capillaries for irradiation and EPR readout. Radiation doses ranging from 1 to 64 Gy were applied with a clinical 6 MV flattening filter free photon beam. EPR readout was then performed with a X-band benchtop spectrometer. The dose response, temporal stability and reproducibility of the samples' EPR signal amplitudes as well as the influence of the nitroxide concentration between 10 and 160μM on the absolute signal loss were investigated using MmP. CmP was used to examine the dependence of the dose response on DMSO concentration between 0 and 10 vol%. An indirect effect model was fitted to the experimental data assuming irradiation induced radical reactions as the underlying mechanism.Main results. For an initial MmP concentration of 20μM, absolute EPR signal loss is linear up to a dose of 16 Gy with a yield G(-NO·) of approximately 0.4μmol J-1. Within five weeks upon sample irradiation to doses between 0 and 32 Gy relative EPR signal fluctuations were on average (126 readouts) below 1% (1σ). For c(MmP) ≥ 20μM, absolute signal loss is only weakly dependent on c(MmP), whereas it increases strongly with increasing c(DMSO) in the range 0-5 vol%. An indirect effect model is applicable to describe the reaction mechanism resulting in the observed dose response curve.Significance. Liquids consisting of nitroxides in aqueous solution and small amounts of DMSO (2 vol%) show promising basic characteristics for application as water-equivalent EPR dosimeter materials in radiotherapy. The EPR signal loss is based on an indirect effect mediated by diffusing radicals originating from the radiolysis of the water/DMSO mixture.

Effect of 1-(hydroxygermatrane monohydrate) on rate of healing of skin burn wounds

It has been experimentally established that when treating burn wounds of the skin by applying a solution of 1-(hydroxygermatrane monohydrate) at a concentration of 10 vol% in a 30% aqueous solution of dimethyl sulfoxide, an increase in the intensity of proliferative and reparative processes is ensured in the conditions of correction of burn healing skin wounds. In particular, the healing of a burn lesion with applications of 10 vol% 1-(hydroxygermatrane monohydrate) in a 30% solution of dimethyl sulfoxide (DMSO) occurs significantly 46.6% faster than with the use of 30% DMSO alone.Accelerated healing of a burn lesion with applications of 10 vol% 1-(hydroxygermatranemonohydrate) in a 30% solution of dimethyl sulfoxide (DMSO) leads to a multiplereduction, even to zero, in the number of post-burn keloid scars.

Liquid-Vapor Interface of Aqueous Ethylene Glycol Solutions: A Molecular Dynamics Study.

While the organic constituent in an aqueous binary solution enriches its liquid-vapor (l-v) interface, the extent of enrichment can depend nonlinearly on its mole fraction. A microscopic quantification and rationalization of this behavior are crucial to understand the dependence of properties such as surface tension and evaporation rate of the solution on its composition. Extensive all-atom molecular dynamics simulations of aqueous ethylene glycol (EG) solutions show that the composition of the solution at the l-v interface deviates the most from that in the bulk solution at an EG mole fraction of 0.3. The population of EG molecules with their central C-C dihedral in the gauche conformation was found to be higher at the l-v interface than that in the bulk solution to facilitate the orientation of its hydrophobic methyl groups toward the vapor phase. Free energy calculations reveal that in dilute EG solutions, an EG molecule is most stable at the l-v interface. The behavior of vapor pressure in aqueous EG solutions is ideal and follows Raoult's law, while in contrast, the aqueous solution of dimethyl sulfoxide does not. A rationale for the same is provided through the orientational distribution of interfacial water molecules in the respective solutions.

Solubility and Thermodynamics Data of Cabozantinib Malate in Various Aqueous Solutions of Dimethyl Sulfoxide at Different Temperatures

Cabozantinib malate (CBZM), a new anticancer medication, has been studied for its solubility and thermodynamic properties in a variety of {dimethyl sulfoxide (DMSO) + water (H2O)} mixtures at 298.2–318.2 K and 101.1 kPa. Using the shake flask technique, the solubility of CBZM was assessed and the results were correlated to the van’t Hoff, Apelblat, Buchowski–Ksiazczak λh, Yalkowsky–Roseman, Jouyban–Acree, and Jouyban–Acree-van’t Hoff models. There was a significant correlation between the experimental CBZM solubility data and all computational models, as evidenced by the error values for all computational models being less than 5.0%. Temperature and DMSO mass percentage improved the CBZM mole fraction solubility in the cosolvent solutions of {DMSO + H2O}. At 318.2 K, pure DMSO had the highest mole fraction solubility of CBZM (4.38 × 10−2), whereas pure H2O had the lowest mole fraction solubility (2.24 × 10−7 at 298.2 K). The positive values of computed thermodynamic parameters indicated that the dissolution of CBZM was endothermic and entropy-driven in all of the {DMSO + H2O} solutions investigated. It was found that the CBZM solvation in {DMSO + H2O} solutions is governed by enthalpy. When compared to CBZM-H2O, CBZM-DMSO showed the highest molecular interactions. The findings of this investigation demonstrated that DMSO has a great deal of potential for CBZM solubilization in H2O.

The Study of the Effect of Dimethylsulfoxide (or Diethylsulfoxide) on Quinine Sulfate-DNA Binding by UV-Vis and Steady-State Fluorescence Spectroscopies.

The effect of dimethylsulfoxide (DMSO) and diethylsulfoxide (DESO) on binding between quinine sulfate (QS) and DNA was studied by virtue of UV-Vis absorption, steady-state fluorescence spectroscopies, and fluorescence polarization measurements. The binding constant was determined at three different temperatures and the values of standard Gibbs energy change, enthalpy and entropy of binding were determined. The mechanism of binding and the effect of sulfoxides on this process was revealed. The values of binding constant, fluorescence polarization and iodide quenching studies confirmed that the main binding mode in QS-DNA system is groove binding. Addition of sulfoxides does not change the binding mechanism. Moreover, with addition of sulfoxides binding constant increases due to the removal of water molecules from DNA grooves making them more available for QS molecules.To explain the effect of DMSO and DESO on QS-DNA binding the photophysical properties of QS in aqueous solutions of DMSO and DESO were also studied. On the basis of quantum yield of QS in water, DMSO and DESO the types of intermolecular interactions were discussed. The obtained results show that quantum yield of QS in sulfoxides is lower compared with that in water and aqueous solution of 0.1M H2SO4. QS forms ground state complexes with both DMSO and DESO that are stronger fluorophores compared with free QS molecules.

Oral self-microemulsifying drug delivery system for honokiol's stress responses attenuation and anti-Cryptocaryon irritans efficacy enhancement in Trachinotus ovatus

In this study, an oral self-micro-emulsifying drug delivery system of honokiol (HN-SMEDDS) was developed to overcome honokiol's disadvantages, such as poor aqueous solubility and induced stress responses. The safeties of HN-SMEDDS and honokiol dimethyl sulfoxide aqueous solution (HN-DMSO) were evaluated by comparing their hemolytic activity and acute toxicity in pompano (Trachinotus ovatus). The one-h-ten-percent hemolytic concentration (EC10) of HN-SMEDDS on pompano erythrocytes was 1.206 mg/mL, 33.5 times that of HN-DMSO. And the minimum lethal doses (MLD) for HN-SMEDDS and HN-DMSO in pompano were 1000 mg/kg and 500 mg/kg, respectively. The 14-d relative percent survival (RPS) was calculated after orally administering substance HN-SMEDDS at various doses in pompano cryptocaryoniasis, revealing an optimal dose of 400 mg/kg. To assess efficacy, a comparison was carried out between the oral administration of HN-SMEDDS and HN-DMSO in treating pompano cryptocaryoniasis. This comparison was based on the 14-d RPS and the quantification of C. irritans trophonts in the right second gill (NCTG). In the challenged pompanos treated with HN-SMEDDS and HN-DMSO, the 14-d RPS and 7-d NCTG reached 68.89 ± 3.14% and 6.3 ± 2.1 individuals, and 53.33 ± 0.00% and 30.7 ± 2.1 individuals, respectively. Furthermore, stress-related markers, encompassing mRNA levels of cyp1a, hsp70, gst, and sod in the liver, spleen, and kidney, along with activities of ACP, CAT, SOD, GSH, and T-AOC in these organs and serum, were compared between the pompanos administered with HN-SMEDDS and HN-DMSO. The impact of HN-SMEDDS on the mRNA levels of cyp1a in the liver and kidney, hsp70 in all the analyzed tissues, gst in the liver and spleen, sod in the spleen, as well as the activities of ACP and CAT in liver, spleen, and serum, GSH content in spleen and kidney, SOD in the liver, kidney, and serum, and the T-AOC in liver, spleen, kidney, and serum, were significantly smaller than those of HN-DMSO. Compared to HN-DMSO, HN-SMEDDS could markedly reduce the stress responses of fish and enhance the efficacy of honokiol.

Crystallization and vitrification of solutions with weak intermolecular interaction of the components

An experimental study of the phenomenon of cluster crystallization and vitrification of solutions with weak intermolecular interaction of the components was performed with use of a purposely developed method of microvolumetric scanning tensodilatometry. Volumetric effects accompanying cooling and subsequent heating of aqueous solutions of glycerol and dimethyl sulfoxide were recorded within temperature range 300…135 K, and they are completely in line with patterns of cluster crystallization. The same phenomenon was used to analyze the structure of amorphous fractions of such solutions. Formation inside of them of ice nanocrystals without breakage of hydrogen bonds between water molecules and molecules of cryoprotectant dramatically reduces stability of amorphous fractions. Based on the resulted tensodilatograms, state diagrams for solutions, which include cluster phase, were for the first time plotted.

Thiol-reactive Pyrene Dimer Having A Disulfide Linkage as A Ratiometric Fluorescence Probe to Thiol-containing Biomolecules

A disulfide-linked pyrene dimer (1), which was constructed with two pyrene derivatives and a cystine-derived disulfide linker, was synthesized as a thiol-reactive ratiometric fluorescence probe. 1 alone showed strong excimer fluorescence at around 490 nm in an aqueous dimethyl sulfoxide solution. Upon addition of thiol-containing biomolecules such as l-cysteine, l-homocysteine, and glutathione, the excimer fluorescence of 1 decreased, while monomeric fluorescence increased, reflecting reduction-responsive cleavage of disulfide bond of 1 to give the corresponding thiol 2.

Interaction of ethylene glycols in aqueous Dimethyl sulfoxide Solutions at different Temperatures

تم قياس الكثافة (ρ) واللزوجة (η) لمحاليل الإيثيلين جليكول في النطاق 0.1 إلى 0.35 مول dm-3 في 2٪ و 5٪ (وزن / وزن) ثنائي ميثيل سلفوكسيد (DMSO) - تم قياس مخاليط الماء عند درجات حرارة مختلفة (298.15303.15) ، 308.15 و 313.15 ك) من هذه البيانات التجريبية ، الحجم المولي الظاهري ϕν ، الحد من الحجم المولي الجزئي ϕºν ، المنحدر Sν ، معاملات Jones-Dole A و B ، طاقة Gibbs الخالية من التنشيط للتدفق اللزج للمحلول ∆G * ، تم حساب المحتوى الحراري ∆H * والإنتروبيا ∆S *. وأظهرت النتائج تفاعلات مذيبات أيونية قوية وتفاعلات أيونية ضعيفة ، وتشير أيضًا إلى أن جلايكول الإيثيلين يعمل كقواطع للبنية.

The role of DMSO and MSM in treatment of tendinopathies affection in equine: A comparative study

Superficial digital flexor tendon (SDFT) injury is a main cause of lameness in polo and racehorses inciting long-term lameness and recovery. This study aimed to compare the efficacy of topical dimethyl sulfoxide (DMSO) and Methylsulfonylmethane (MSM) on bowed tendons in horses. Ten client-owned injured horses, aged between 5-15 years with an average age of 8.5 years and weighing approximately 400-500 kg, suffering from chronic tendinitis were selected and allocated into two groups: group A (DMSO) and group B (MSM). The group A animals (n=5) were treated with 60 mL of topical 90% DMSO aqueous solution, while Group B animals (n=5) received 60 mL of topical 90% MSM aqueous solution on the bowed tendon at days 0, 7, 14, 21, 28, 35, and day 42. Clinical examinations were performed to evaluate the severity of pain and lameness grade. Ultrasound scans of the superficial digital flexor tendon were taken at days 0, 21, and day 42. Images were placed in numerical form, and the fiber alignment score of the fibers (FS) and the echogenicity score (ES) of the superficial digital flexor tendon at the maximum injury zone (MIZ) were recorded. Results showed that pain reduction and lameness significantly (P<0.05) reduced from 21d to 42d in the DMSO animals compared with the MSM animals. The echogenicity score didn’t differ between the two groups at 0d, 21d, and 42d (P>0.05) however, it was significantly declined in group A at 42d than 0d (P<0.05). Fiber alignment in group A was (P<0.01) reduced at 42d compared with group B. This study concluded that group A horses showed very effective and long-term improvements compared to the group B horses.

Site selective analysis of water in hydrogen bond network of aqueous dimethyl sulfoxide solutions by oxygen K-edge X-ray absorption spectroscopy

Hydrogen bond (HB) network in aqueous dimethyl sulfoxide (DMSO) solutions at different concentrations has been observed by oxygen K-edge X-ray absorption spectroscopy with a site selective analysis that separates donor and acceptor sites of water (H2O), where the S=O π* peak in DMSO reflects the donor site of H2O and the 4a1 peak in H2O reflects the acceptor site of H2O, respectively. From the comparison of the XAS experiments with molecular dynamics simulations and inner-shell quantum chemical calculations, we have revealed that the HB network in aqueous DMSO solutions is influenced with not only the HB interaction of the S=O group with the donor site of H2O but also the dipole interaction of the S atom with the acceptor site of H2O, which breaks the HB network between H2O. Four concentration regions were found in the HB network of aqueous DMSO solutions, which would be related to the anomalies of physical properties and solvent effects in chemical and biological reactions.

Structure of Protonated Forms of Cryptand[2.2.2] and Constants of Their Formation in Aqueous Solutions of Dimethyl Sulfoxide

Structure of Protonated Forms of Cryptand[2.2.2] and Constants of Their Formation in Aqueous Solutions of Dimethyl Sulfoxide

Cytopathological characteristics for provoked lesions of squamous epithelia of digestive system in animal models

Purpose of the study. Investigation of morphological characteristics of changes in the epithelium of foregut in experimental animals put under conditions of provoked carcinogenesis.Materials and methods. The method of chronic experiment on animals is applied: 40 female non-linear white rats, which are divided into 4 equal groups (n = 10). The first (I) and second (II) control groups of animals were exposed to mechanical trauma of the oral mucosa with additional application of 0.9 % NaCl solution and 1 % aqueous dimethyl sulfoxide (DMSO) solution with a frequency of 2 times a week. The third group of animals, i.e. the main group (4-NQO), was subjected to mechanical traumatization of the oral mucosa with additional application of 1 % DMSO aqueous solution containing 0.1 mg/ml of 4 nitroquinoline-N-oxide, with a frequency of 2 times a week. The fourth group of animals was intact. On the 1st, 7th, 10th, 14th day from the beginning of the experiment, and then every 14 days, cytological material was taken from the oral mucosa and stained with Papanicolaou stain. To compare the emerging changes, the histological picture of the organs of the foregut was studied.Results. Animals from the main group (4-NQO) showed a lag in body weight gain over the standard variance in the control (I and II) and intact groups. Atypical cells of indeterminate significance appeared in smears starting from 42 days of the experiment. Starting from the 56th day of the experiment, atypical cells (1–2 in the field of view), described in Bethesda terms as low-grade intraepithelial lesions, were detected in smears obtained from the main group of animals (4-NQO). Parabasal cells of the squamous epithelium with atypical large (more than 3 times compared to the reference) nuclei, anisonucleosis, with variable contours of the nuclear membrane within one cell, coarse chromatin were recognized as diagnostically significant.Conclusion. The severity of morphological changes in foregut epithelium in the direction of the precancerous state is significantly higher in the main group with prolonged exposure to carcinogenic factor, compared with the control and intact groups (р &lt; 0.05). Thus, the effectiveness of modeling the conditions of provocation of carcinogenesis of the epithelium of the upper digestive tube in experimental animals was studied and proved by mechanical traumatization of the oral mucosa with additional application of carcinogen (4-NQO). This model of provocation of carcinogenesis will be used in the next study of neoplastic processes of the female reproductive system.

Increasing the ⋅OH radical concentration synergistically with plasma electrolysis and ultrasound in aqueous DMSO solution

In recent years, significant increases in waste processing and material engineering have been seen by using advanced oxidation processes. The treatment results and energy yields of these processes are largely determined by the generation and retention of reactive oxygen species (ROS). However, increasing the amount of ROS remains a key challenge because of the unavailability of performance- and energy-efficient techniques. In this study, plasma electrolysis, ultrasound, and plasma electrolysis combined with ultrasound were used to treat dimethyl sulfoxide (DMSO) solutions, and the results showed that the two methods can synergistically convert filament discharge into spark discharge, and the conversion of the discharge mode can significantly increase the concentration of OH radicals and effectively improve the efficiency of DMSO degradation. We verified the rationality of the results by analyzing the mass transfer path of ROS based on the reaction coefficients and found that the ⋅OH radicals in aqueous solution were mainly derived from the decomposition of hydrogen peroxide. These findings indicated that the synergistic action of plasma electrolysis and ultrasound can enhance the production of chemically reactive species, and provide new insights and guiding principles for the future translation of this combined strategy into real-life applications. Our results demonstrated that the synergistic strategy of ultrasound and plasma electrolysis is feasible in the switching mode and increasing the ROS, and may open new routes for materials engineering and pollutant degradation.

Cytopathological characteristics for provoked lesions of squamous epithelia of the digestive system in animal models

Relevance: Noticeable spreading of malignant neoplasms of the upper section of the digestive system and the worsening of these patients’ quality of life in case of late diagnostics stimulate the development of protocols for early detection of precancerous changes.&#x0D; The purpose of the study was to investigate the morphological characteristics of changes in the squamous epithelium of the upper part of the digestive tube in model organisms under provoked carcinogenesis when exposed to substances with proven carcinogenic activity.&#x0D; Methods: 40 female nonlinear white rats from the Orekhovo-Zuevsky urban settlement veterinary station were used as model organisms. The rats were divided into four equal groups. The first control group was not exposed to any specific influences. In groups 2-4, the rats underwent sedation and mechanical scarification microtraumatization of the oral mucosa. Then, they were applied twice a week by 0.9% NaCl solution (control group 2), 1% aqueous solution of dimethyl sulfoxide (DMSO, Akrikhin, Russia) (control group 3), or 1% aqueous solution of DMSO containing 0.1 mg/ml 4-nitroquinoline-N-oxide (4-NQO, Acros Organics, USA) (Experimental Group 4).&#x0D; Results: The cytological picture associated with chronic trauma to the mucous membrane of the upper digestive tract was characterized by nonspecific reactions: inflammation, hyper- and parakeratosis, and atrophic changes. There were no statistically significant differences in the frequency of such changes between the experimental and control groups. Specific changes in squamous epithelium towards a precancerous state in the Experimental Group with chronic exposure to a substance with carcinogenic activity occurred significantly more often than in the control groups (p &lt;0.05), as proven by the results of comparative histological verification. Conclusion: Cytological study could be used in clinical practice as a screening tool for precancerous oropharynx and upper esophagus conditions.

Evaluation of Drug Sorption on Laboratory Materials with Abraham Solvation Parameters of Drugs and its Prevention.

Undesired drug sorption on laboratory material surfaces reduces the performance of analytical methods and results in the generation of unreliable data. Hence, we characterized the sorption of drugs and evaluated the sorption extent using a linear free energy relationship (LFER) model with Abraham solvation parameters of drugs. Furthermore, to prevent sorption, the effects of additives, such as organic solvents and salts, were evaluated. The sorption of fifteen model drugs (concentration: 2μM), with various physicochemical properties, on materials in 0.2% dimethyl sulfoxide aqueous solutions was evaluated. Drug sorption extent on the materials was determined using high-performance liquid chromatography. The obtained results were analyzed using an LFER model with Abraham solvation parameters of the drugs. The effect of additives on the sorption of itraconazole, one of the most hydrophobic drugs among those tested in this study, was investigated. Sorption was dependent on the physicochemical properties of drugs, rather than the type of materials used, and additives altered the rate of drug sorption. Equations were developed to evaluate the sorption extent (nmol) of drugs to glass and polypropylene using the Abraham solvation parameters of the drugs. LFER modeling with Abraham solvation parameters of drugs enabled us to evaluate drug sorption on materials. All the additives altered the rate of drug sorption, and some organic solvents effectively prevented sorption. The developed LFER model would be useful for assessment of the sorption properties of compounds in in vitro evaluations in drug discovery research and various other biochemical fields.

Enthalpic Characteristics of the Dissociation of Glycylglycine and Glycine in Aqueous Solutions of Dimethylsulfoxide: Calculations and Experiments

Enthalpic Characteristics of the Dissociation of Glycylglycine and Glycine in Aqueous Solutions of Dimethylsulfoxide: Calculations and Experiments

Thermoplastic Analysis of Cluster Crystallization of Cryoprotective Solutions

For the first time the possibility of using thermomechanical analysis to construct the fragments of state diagrams of cryoprotective solutions in the zone of glass transition temperatures has been considered. A method for studying cluster crystallization of cryoprotective solutions based on thermomechanical curves has been developed. The parameters of thermomechanical curves of frozen aqueous solutions of dimethyl sulfoxide (DMSO), polyethylene oxide 1500 (PEO-1500), glycerol were analyzed and the relationship between these parameters and the cluster crystallization kinetics for these solutions was established. On the basis of experimentally obtained thermomechanical curves for the frozen solutions of DMSO and PEO-1500 the possibility of formation of clusters of two types has been shown: on the basis of ice and cryoprotective substance microcrystals. Additional experimental data were obtained to construct a complete state diagrams of cryoprotective solutions, which include the existing cluster phase areas.

Evaluation of acute and subacute toxicity of ethanolic extract and fraction of alkaloids from bark of Aspidosperma nitidum in mice

This study investigated the acute and subacute toxicity of the ethanolic extract (EE) and alkaloid fraction (FA) from A. nitidum. The EE was obtained from trunk bark with ethanol, FA was obtained from the fractionation of EE. To test the acute toxicity, mice were divided into four groups, and the negative controls received water or aqueous solution of dimethyl sulfoxide, whereas the others received EE or FA (2000 mg/kg, orally, single dose). The same controls were used in the subacute trial. However, the animals were treated for 28 days, and the dose used was 1000 mg/kg per day of EE and FA. Daily clinical evaluations of the animals were performed. At the end of the experiment, hematological, biochemical, and histopathological assessments (liver, lung, heart, and kidney) were performed. In the acute and subacute toxicity studies, mice treated with EE and FA did not show any clinical changes, there were no changes in weight gain, hematological and biochemical parameters compared to the control groups (p > 0.05). In the histopathological examination, there was no abnormality in the organs of the treated animals. Therefore, EE and FA did not produce toxic effects in mice after acute and subacute treatment.