Short-term test for the toxicogenomic assessment of ecotoxic modes of action in Myriophyllum spicatum.

In environmental risk assessment of substances, the 14-day growth inhibition test following OECD test guideline 239 is employed to assess toxicity in the macrophyte Myriophyllum spicatum. Currently, this test evaluates physiological parameters and does not allow the identification of the mode of action (MoA) by which adverse effects are induced. However, for an improved ecotoxicity assessment of substances, knowledge about their ecotoxic MoA in non-target organisms is required. It has previously been suggested that the identification of gene expression changes can contribute to MoA identification. Therefore, we developed a shortened three-day assay for M. spicatum including the transcriptomic assessment of global gene expression changes and applied this assay to two model substances, the herbicide and photosynthesis inhibitor bentazone and the pharmaceutical and HMG-CoA reductase inhibitor atorvastatin. Due to the lack of a reference genome for M. spicatum we performed a de novo transcriptome assembly followed by a functional annotation to use the toxicogenomic results for MoA discrimination. The gene expression changes induced by low effect concentrations of these substances were used to identify differentially expressed genes (DEGs) and impaired biological functions for the respective MoA. We observed both concentration-dependent numbers and differentiated patterns of DEGs for both substances. While bentazone impaired genes involved in the response to reactive oxygen species as well as light response, and also genes involved in developmental processes, atorvastatin exposure led to a differential regulation of genes related to brassinosteroid response as well as potential metabolic shifts between the mevalonate and methyl erythritol 4-phosphate pathway. Based on these responses, we identified biomarker candidates for the assessment of MoA in M. spicatum. Utilizing the shortened assay developed in this study, the investigation of the identified biomarker candidates may contribute to the development of future MoA-specific screening approaches in the ecotoxicological hazard prediction using aquatic non-standard model organisms.

Investigating the impacts of airborne dust on herbicide performance on Amaranthus retroflexus.

Dust pollution poses environmental hazards, affecting agriculture through reduced sunlight exposure, photosynthesis, crop yields, and food security. This study explores the interference of dust pollution on herbicide efficacy to control weeds in a semi-arid region. In a factorial experiment conducted in 2019 and replicated in 2020, the interaction of dust and various herbicide applications, including bentazon, sulfosulfuron, tribenuron-methyl, aminopyralid + florasulam, foramsulfuron + iodosulfuron + thiencarbazone, 2,4-D + MCPA, and acetochlor, in controlling Amaranthus retroflexus L. were assessed. Dust induced a 9.2% reduction in the total chlorophyll content of A. retroflexus, while herbicide application independently led to a 67.5% decrease. Contrary to expectations, herbicides performed better in dust, except bentazon, which caused a 28% drop in plant height and a 29% decrease in total biomass compared to non-dust conditions. Both herbicides and dust exerted suppressive effects on A. retroflexus's leaf and stem weights and overall biomass. Despite dust presence, tribenuron-methyl (95.8%), aminopyralid + florasulam (95.7%), sulfosulfuron (96.5%), and foramsulfuron + iodosulfuron + thiencarbazone (97.8%) effectively controlled A. retroflexus. These findings indicate that dust's effect on herbicide efficacy is herbicide-dependent but except bentazon, dust generally increased herbicide efficacy and amplified the control of A. retroflexus.

Zero-waste preparation of mixed oxides for submicromolar sensing of Bentazone pesticide.

Necessity of having simple selective and robust methods for the analysis of environmentally relevant chemicals stimulates the development of new approaches to material preparation. Electrochemical sensing using electroactive substrates has proved efficient in the analysis of a wide range of pesticides and is widely used as a routine analytical method. Recently, mixed oxides showed promising electrocatalytic activity toward hazardous substrates. Prevalence of wet chemical methods in the synthesis of mixed oxides creates a methodological obstacle and inconvenience for their wide utilization. In this work we challenged the common preparation of mixed oxides with simple powder mixing and developed an electrode for bentazone detection with satisfactory detection limit (0.4 µM), recovery rate (≈104%), and a broad linearity range (1-45 µM). The proposed modified carbon paste electrode is highly selective and can be used for determination of bentazone in presence of interfering ions in water samples.

Discovery of a Novel Benzothiadiazine-Based Selective Aldose Reductase Inhibitor as Potential Therapy for Diabetic Peripheral Neuropathy.

Aldose reductase 2 (ALR2), an activated enzyme in the polyol pathway by hyperglycemia, has long been recognized as one of the most promising targets for complications of diabetes, especially in diabetic peripheral neuropathy (DPN). However, many of the ALR2 inhibitors have shown serious side effects due to poor selectivity over aldehyde reductase (ALR1). Herein, we describe the discovery of a series of benzothiadiazine acetic acid derivatives as potent and selective inhibitors against ALR2 and evaluation of their anti-DPN activities in vivo. Compound 15c, carrying a carbonyl group at the 3-position of the thiadiazine ring, showed high potent inhibition against ALR2 (IC50 = 33.19 nmol/L) and ∼16,109-fold selectivity for ALR2 over ALR1. Cytotoxicity assays ensured the primary biosafety of 15c. Further pharmacokinetic assay in rats indicated that 15c had a good pharmacokinetic feature (t1/2 = 5.60 h, area under the plasma concentration time curve [AUC(0-t)] = 598.57 ± 216.5 µg/mL * h), which was superior to epalrestat (t1/2 = 2.23 h, AUC[0-t] = 20.43 ± 3.7 µg/mL * h). Finally, in a streptozotocin-induced diabetic rat model, 15c significantly increased the nerve conduction velocities of impaired sensory and motor nerves, achieved potent inhibition of d-sorbitol production in the sciatic nerves, and significantly increased the paw withdrawal mechanical threshold. By combining the above investigations, we propose that 15c might represent a promising lead compound for the discovery of an antidiabetic peripheral neuropathy drug.

Embryonic Development Effects of Basagran® Herbicide in Danio Rerio: A Preliminary Study.

Bentazon (Basagran®) belongs to the chemical group of benzothiadiazinones. Thus, this study aimed to estimate the influence of herbicide bentazon (3 µg.L-1, 6 µg.L-1, 12 µg.L-1, 300 µg.L-1) in Danio rerio embryos development. The study tested environmental relevant concentrations of bentazon as well as the limit established for drinking water (300 µg.L-1) in Brazil. We performed behavioral and developmental analyzes during 96 h of exposure. The bentazon measurements after experimental period showed reduction ranging from 5.0 to 18.93% between exposed groups. Our results showed significant differences in the heart rate, which was significantly higher in groups exposed to all bentazon concentrations compared to control groups. The absence of alterations in the behavioral parameters showing that the herbicide bentazon at the concentrations tested had few adverse effects on the development and behavior of the Danio rerio embryos. Considering the toxic point of view, there is a chance that bentazon acts together with other environmental contaminants as an additive or synergistic way.

Multiple Metabolism Pathways of Bentazone Potentially Regulated by Metabolic Enzymes in Rice.

Bentazone (BNTZ) is a selective and efficient herbicide used in crop production worldwide. However, the persistence of BNTZ residues in the environment has led to their increasing accumulation in farmland and crops, posing a high risk to human health. To evaluate its impact on crop growth and environmental safety, a comprehensive study was conducted on BNTZ toxicity, metabolic mechanism, and resultant pathways in rice. The rice growth was compromised to the treatment with BNTZ at 0.2-0.8 mg/L (529.95-1060.05 g a.i./ha), while the activities of enzymes including SOD, POD, CAT, GST, GT, and CYP450 were elevated under BNTZ stress. A genome-wide RNA-sequencing (RNA-Seq) was performed to dissect the variation of transcriptomes and metabolic mechanisms in rice exposed to BNTZ. The degradative pathways of BNTZ in rice are involved in glycosylation, hydrolysis, acetylation, and conjugation processes catalyzed by the enzymes. Our data provided evidence that helps understand the BNTZ metabolic and detoxic mechanisms.

New insights in the development of positive allosteric modulators of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors belonging to 3,4-dihydro-2H-1,2,4-benzothiadiazine 1,1-dioxides: Introduction of (mono/difluoro)methyl groups at the 2-position of the thiadiazine ring.

Positive allosteric modulators of the AMPA receptors (AMPAR PAMs) have been proposed as new drugs for the management of various neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, attention deficit hyperactivity disorder, depression, and schizophrenia. The present study explored new AMPAR PAMs belonging to 3,4-dihydro-2H-1,2,4-benzothiadiazine 1,1-dioxides (BTDs) characterized by the presence of a short alkyl substituent at the 2-position of the heterocycle and by the presence or absence of a methyl group at the 3-position. The introduction of a monofluoromethyl or a difluoromethyl side chain at the 2-position instead of the methyl group was examined. 7-Chloro-4-cyclopropyl-2-fluoromethyl-3,4-dihydro-4H-1,2,4-benzothiadiazine 1,1-dioxide (15e) emerged as the most promising compound associating high in vitro potency on AMPA receptors, a favorable safety profile in vivo and a marked efficacy as a cognitive enhancer after oral administration in mice. Stability studies in aqueous medium suggested that 15e could be considered, at least in part, as a precursor of the corresponding 2-hydroxymethyl-substituted analogue and the known AMPAR modulator 7-chloro-4-cyclopropyl-3,4-dihydro-4H-1,2,4-benzothiadiazine 1,1-dioxide (3) devoid of an alkyl group at the 2-position.

Design, Synthesis, and Evaluation of Novel Pyruvate Dehydrogenase Kinase Inhibitors.

AIMS: The present work describes the synthesis and the biological evaluation of novel compounds acting as pyruvate dehydrogenase kinase (PDK) inhibitors. These drugs should become a new therapeutic approach for the treatment of pathologies improved by the control of the blood lactate level. METHODS: Four series of compounds belonging to N-(4-(N-alkyl/aralkylsulfamoyl)phenyl)-2- methylpropanamides and 1,2,4-benzothiadiazine 1,1-dioxides were prepared and evaluated as PDK inhibitors. RESULTS: The newly synthesized N-(4-(N-alkyl/aralkylsulfamoyl)phenyl)-2-methylpropanamides structurally related to previously reported reference compounds 4 and 5 were found to be potent PDK inhibitors (i.e. 10d: IC50 = 41 nM). 1,2,4-Benzothiadiazine 1,1-dioxides carrying a (methyl/ trifluoromethyl)-propanamide moiety at the 6-position were also designed as conformationally restricted ring-closed analogues of N-(4-(N-alkyl/aralkylsulfamoyl)phenyl)-2-hydroxy-2-methylpropanamides. Most of them were found to be less potent than their ring-opened analogues. Interestingly, the best choice of hydrocarbon side chain at the 4-position was the benzyl chain, providing 11c (IC50 = 3.6 µM) belonging to "unsaturated" 1,2,4-benzothiadiazine 1,1-dioxides, and 12c (IC50 = 0.5 µM) belonging to "saturated' 1,2,4-benzothiadiazine 1,1-dioxides. CONCLUSION: This work showed that ring-closed analogues of N-(4-(N-alkyl/aralkylsulfamoyl) phenyl)- 2-hydroxy-2-methylpropanamides were less active as PDK inhibitors than their corresponding ring-opened analogues. However, the introduction of a bulkier substituent at the 4-position of the 1,2,4-benzothiadiazine 1,1-dioxide core structure, such as a benzyl or a phenethyl side chain, was allowed, opening the way to the design of new inhibitors with improved PDK inhibitory activity.

Tandem Reduction, Ammonolysis, Condensation, and Deamination Reaction for Synthesis of Benzothiadiazines and 1-(Phenylsulfonyl)-1H-benzimidazoles.

A novel route for a SnCl2-promoted tandem reduction, ammonolysis, condensation, and deamination reaction which uses nitrile and 2-nitro-N-phenylbenzenesulfonamide/N-(2-nitrophenyl)benzenesulfonamide to synthesize derivatives of benzothiadiazine/1-(phenylsulfonyl)-1H-benzimidazole has been developed. The method features convenient operation and good functional group tolerance. In addition, it employs unsensitive and inexpensive SnCl2/i-PrOH as the reaction reagent and provides a direct approach for the synthesis of pharmaceutically important targets.

National Assessment of Long-Term Groundwater Response to Pesticide Regulation.

Quantitative assessments of long-term, national-scale responses of groundwater quality to pesticide applications are essential to evaluate the effectiveness of pesticide regulations. Retardation time in the unsaturated zone (Ru) was estimated for selected herbicides (atrazine, simazine, and bentazon) and degradation products (desethylatrazine (DEA), desisopropylatrazine (DIA), desethyldesisopropylatrazine (DEIA), and BAM) using a multidecadal time series of groundwater solute chemistry (∼30 years) and herbicide sales (∼60 years). The sampling year was converted to recharge year using groundwater age. Then, Ru was estimated using a cross-correlation analysis of the sales and the frequencies of detection and exceedance of the drinking water standard (0.1 µg/L) of each selected compound. The results showed no retardation of the highly polar, thus mobile, parent compounds (i.e., bentazon), while Ru of the moderately polar compounds (i.e., simazine) was about a decade, and their degradation products showed even longer Ru. The temporal trends of the degradation products did not mirror those of the sale data, which were attributed to the various sale periods of the parent compounds, sorption of the parent compounds, and complex degradation pathways. The longer Ru in clayey/organic sediments than in sandy sediments further confirmed the role of soil-specific retardation as an important factor to consider in groundwater protection.

Investigation of resistance mechanisms to bentazone in multiple resistant Amaranthus retroflexus populations.

Redroot amaranth (Amaranthus retroflexus L.) is a noxious weed that affects soybean production in China. Experiments were conducted to determine the molecular basis of resistance to bentazone. Whole-plant dose-response experiments showed that two populations (R1 and R2) exhibited resistance to bentazone with resistance indices of 9.01 and 6.85, respectively. Sequencing of the psbA gene revealed no amino acid substitution in the two populations. qRT-PCR analysis verified that psbA gene expression in R1 and R2 populations was increased significantly after treatment with bentazone, which was 3-fold and 5-fold higher than that in S1 and S2 populations, respectively. The P450 inhibitor malathion significantly reduced the level of resistance in the R1 and R2 populations when used prior to bentazone treatment. The R1 population exhibited multiple resistance to thifensulfuron-methyl and lactofen, caused by target site mutations (Asp-376-Glu in ALS, Arg-128-Gly in PPO2). In conclusion, increased gene expression of the psbA gene and enhanced herbicide metabolism seem to be the basis of resistance to bentazone in these A. retroflexus populations.

Pulsed-sonochemiluminescence combined with molecularly imprinted polymerized high internal phase emulsion adsorbent for determination of bentazone.

A small low-power humidifier with a simple programmable on/off switch was used as a pulsed ultrasound generator. Using this tool, a novel sonochemiluminescence (SCL) method was developed to determine bentazone. To the best of our knowledge, no chemiluminescence method has been proposed to determine this pesticide. Only five studies have been proposed for SCL quantitative applications so far. Therefore, revealing new aspects of SCL promises to develop analytical methods for the quantitative determination of different substances. A molecularly imprinted polymerized high internal phase emulsion (MIP-polyHIPE) was synthesized, bentazone separated from aqueous solutions, and pre-concentrated by the MIP-polyHIPE foam. The adsorption of bentazone on the MIP-polyHIPE adsorbent was theoretically studied by density functional theory through molecular dynamics simulation. Both experimental and simulation results indicated removal and pre-concentration of bentazone by the MIP-polyHIPE adsorbent. Using the proposed SCL method and without pre-concentration process, a linear dynamic range (LDR) of 2.5 × 10-7-5.0 × 10-5 mol L-1 and a limit of detection (LOD) of 8.4 × 10-8 mol L-1 were obtained for bentazone with a relative standard deviation of 2.64%. The LDR and LOD were improved to 2.6 × 10-9-2.0 × 10-7 mol L-1 and 8.8 × 10-10 mol L-1, respectively, using MIP-polyHIPE adsorbents. The method's application was evaluated by removing and pre-concentration of bentazone from water samples, including well, river, and tap water. The results showed that the pre-concentration factor and recovery percentages were 113-131 times and 93-106%, respectively, using the MIP-polyHIPE absorbent.

Modeling of Bentazone Leaching in Soils with Low Organic Matter Content.

The aim of this study was to estimate bentazone's potential to leach to groundwater in the Arenosols developed from sand, Luvisols developed from loamy sand or sandy loam, and Luvisols or Cambisols developed from loess, and to identify the major factors influencing bentazone's fate in the soils. Potato and maize cultivations were simulated using the FOCUS PELMO 5.5.3 pesticide leaching model. The amount of bentazone reaching groundwater was highly sensitive to degradation parameters, water-holding capacity, evapotranspiration, organic carbon content, and pH. The highest bentazone concentrations in percolate were noted in Arenosols. The risk of bentazone concentration exceeding 0.1 µg/L was low only in Arenosols with high organic carbon content (3.0% for topsoil or higher). In Luvisols developed from loamy sand or sandy loam, the estimated bentazone concentrations in percolate were highly dependent on the climate. In Luvisols or Cambisols developed from loess, concentrations of >0.1 µg/L were the least likely due to the high water-holding capacity and high organic carbon content of these soils. The study also revealed that the FOCUS Hamburg scenario, representing the coarsest soils in the European Union with relatively low organic carbon content, does not reflect the leaching potential of Arenosols and Luvisols.

Cobalt(III)/Chiral Carboxylic Acid-Catalyzed Enantioselective Synthesis of Benzothiadiazine-1-oxides via C-H Activation.

Among sulfoximine derivatives containing a chiral sulfur center, benzothiadiazine-1-oxides are important for applications in medicinal chemistry. Here, we report that the combination of an achiral cobalt(III) catalyst and a pseudo-C2 -symmetric H8 -binaphthyl chiral carboxylic acid enables the asymmetric synthesis of benzothiadiazine-1-oxides from sulfoximines and dioxazolones via enantioselective C-H bond cleavage. With the optimized protocol, benzothiadiazine-1-oxides with several functional groups can be accessed with high enantioselectivity.

Chlorinated benzothiadiazines inhibit angiogenesis through suppression of VEGFR2 phosphorylation.

Angiogenesis inhibitors are a critical pharmacological tool for the treatment of solid tumors. Suppressing vascular permeability leads to inhibition of tumor growth, invasion, and metastatic potential by blocking the supply of oxygen and nutrients. Disruption of the vascular endothelial growth factor (VEGF) signaling pathway is a validated target for the design of antiangiogenic agents. Several VEGFR2 inhibitors have been clinically approved over the past years. Structural analysis of these clinical VEGFR2 inhibitors highlighted key functional group overlap with the benzothiadiazine core contained in a library of in-house compounds. Herein we ascribe anti-angiogenic activity to a series of chlorinated benzothiadiazines. Selected compounds show significant activity to completely ameliorate VEGF-induced endothelial cell proliferation by suppression of VEGFR2 phosphorylation. The scaffold is devoid of activity to inhibit carbonic anhydrases and generally lacks cytotoxicity across a range of cancer and non-malignant cell lines. Assay of activity at 468 kinases shows remarkable selectivity with only four kinases inhibited > 65% at 10 µM concentration, and with significant activity to inhibit TNK2/ACK1 and PKRD2 by > 90%. All four identified kinase targets are known modulators of angiogenesis, thus highlighting compound 17b as a novel angiogenesis inhibitor for further development.

Benzothiadiazines derivatives as novel allosteric modulators of kainic acid receptors.

The majority of excitatory neurotransmission in vertebrate CNS is mediated by glutamate binding to different types of receptors. Among them, a-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) and kainite receptors (KAR) are ionotropic receptors playing important pathophysiological roles. A number of small molecules acting as positive allosteric modulators (PAM) of AMPAR have been proposed as drugs for neurological disorders, however, there is no such abundance of ligands capable of modulating KARs activity. We investigated the ability of IDRA21 and of its derivative, compound 2 (c2), to modulate glutamate-evoked currents at native and recombinantly expressed AMPA and KA receptors. By using the patch clamp technique we analyzed the activity of the two compounds in primary cultures of cerebellar granule neurons and in HEK293 cells transiently transfected with KARs and AMPAR subunits. It resulted that both benzothiadiazine derivatives potentiate AMPAR and KAR mediated currents in native and recombinant receptors, c2 being always more potent and efficacious than IDRA21. The potency of both compounds was higher in native receptors than in recombinant receptors. In HEK293 cells transfected with AMPAR subunits, the efficacy of IDRA21 and c2 was much higher in GluA1 than in GluA2 homomeric receptors while their potency did not change. In recombinant KAR, both compounds had a potency in the high micromolar range, while the efficacy reached a maximum in the GluK2 expressing cells. The benzothiadiazine effect, both in native and recombinant receptors, was detected mainly on plateau current, involving a decrease in AMPAR and KAR desensitization. Our study demonstrates for the first time that two positive allosteric modulators of AMPAR, IDRA21 and its derivative, c2, potentiate KAR activity. Furthermore, we highlighted their subunit selectivity that may enable the design of potent and selective PAMs, which could be relevant for the development of new drugs and for a better understanding of KAR functions in the CNS.

Automation in quantifying phenoxy herbicides and bentazon in surface water and groundwater using novel solid phase extraction and liquid chromatography tandem mass spectrometry.

The occurrence of phenoxy herbicides is a financial and regulatory concern for drinking water treatment plants. This paper presents a new method of quantification for nine phenoxy-acids and bentazon in different water samples using liquid chromatography tandem mass spectrometry (LC-MS/MS). The method is based on an automated solid phase extraction (SPE) process that applied hydrophilic modified polystyrene and divinylbenzene cartridges at low pH (<2.0). Main advantages of the presented method include the reduced consumption of organic solvent in extraction and the fully automated sample pre-concentration. The method is thus more environmentally-friendly. In the quantification step, five stable isotopically labelled analogues were used as internal standards to account for the losses during sample preparation and to calibrate the ion source response under the mass spectrometric detection. The method was optimized in terms of sample preparation and subsequent LC-MS/MS separation to obtain reliable measurement of the analyte concentration during real sample analysis. The method quantification limit was between 1.5 and 10.0 ng/L for target compounds in surface water and groundwater samples. The method was validated at three fortification levels between 10.0 and 1000 ng/L, and the results showed fit-for-purpose recovery with appropriate precision at low concentration levels. The method was also utilized to analyse thirty-two actual water samples from different sources. Forty percent of the analysed samples contained detectable level of herbicides, ranging from 1.91 to 40.5 ng/L. The concentrations of targeted herbicides in our study were comparable to those found in water samples in other regions of world.

Bentazone in water and human urine in Wuhan, central China: exposure assessment.

Bentazone is a widely used post-emergence herbicide, while no data was available on its concentrations in tap water from China and in urine among the general population. It was determined in the source (Wuhan section of the Yangtze River watershed), treated, and tap water (n = 20, 20, and 170, respectively) in different seasons (2019) in Wuhan, central China. Also, urine samples (n = 38) collected from healthy adults in Wuhan (September 2020) were analyzed to characterize its urinary concentration. Bentazone was detected in all the source and treated water samples. Its concentrations in the source water in July were higher than those in February (median: 17.9 ng/L vs. 2.86 ng/L) (p < 0.05). It cannot be removed efficiently (27.8-27.9%) by conventional drinking water treatment using NaClO, but it can be efficiently removed by using chlorine dioxide or ozone combined with activated carbon. Bentazone was frequently detected (detection frequency: 96.3%) in 160 tap water samples (underwent conventional treatment) (median: 1.95 ng/L, range: <0.02-47.0 ng/L), while it was not detectable in tap water samples that underwent ozone combined with activated carbon. Seasonal variations were found, with the lowest median concentration (ng/L) in April (0.46) and the highest in July (17.6). In addition, bentazone was frequently (92.1%) detected in human urine samples (median: 0.02 ng/mL; range: < 0.01-0.11 ng/mL). The estimated daily intake of bentazone based on its median concentration in tap water (0.04 ng/kg-body weight [bw]/day) accounted for approximately 8% of that based on the median urinary concentration (0.48 ng/kg-bw/day). This is the first time to characterize its occurrence in drinking water from China and its occurrence in the urine of the general population.

THE EFFECT OF ADEMOL ON THE DNA FRAGMENTATION OF CEREBRAL CORTEX CELLS IN RATS WITH EXPERIMENTAL TRAUMATIC BRAIN INJURY.

THE AIM OF THE STUDY: To determine the effect of Ademol on the deoxyribonucleic acid (DNA) fragmentation of the cerebral cortex cells (apoptosis) of rats with traumatic brain injury (TBI). MATERIALS AND METHODS: An experimental model of severe trauma was created in male rats using an air pistol. The therapeutic effect of Ademol in TBI was evaluated at a dose of 2 mg/kg intravenously at intervals of 2 t/d for 8 days. As a drug for the control group we used 0.9% NaCl at a dose of 2 ml/kg, and as a comparison drug - amantadine sulfate at a dose of 5 mg/kg. On day 8 after TBI and decapitation of animals, the parts of the cerebral cortex were taken to assess further DNA fragmentation in cells by the flow cytometry method. RESULTS AND CONCLUSIONS: The post-traumatic period of model TBI in rats is accompanied by a probable increase in the level of DNA fragmentation in the nucleus of cerebral cortex cells on the 8th day of the experiment. By the antiapoptotic effect in conditions of post-traumatic brain injury, Ademol solution therapy was significantly better than the infusion of 0.9% NaCl and amantadine sulfate at an average of 46.2 and 27.2%, respectively (p<0.05).

Structure of benzothiadiazine at zwitterionic phospholipid cell membranes.

The use of drugs derived from benzothiadiazine, which is a bicyclic heterocyclic benzene derivative, has become a widespread treatment for diseases such as hypertension (treated with diuretics such as bendroflumethiazide or chlorothiazide), low blood sugar (treated with non-diuretic diazoxide), or the human immunodeficiency virus, among others. In this work, we have investigated the interactions of benzothiadiazine with the basic components of cell membranes and solvents, such as phospholipids, cholesterol, ions, and water. The analysis of the mutual microscopic interactions is of central importance to elucidate the local structure of benzothiadiazine as well as the mechanisms responsible for the access of benzothiadiazine to the interior of the cell. We have performed molecular dynamics simulations of benzothiadiazine embedded in three different model zwitterionic bilayer membranes made by dimyristoylphosphatidylcholine, dioleoylphosphatidylcholine, 1,2-dioleoyl-sn-glycero-3-phosphoserine, and cholesterol inside aqueous sodium-chloride solution in order to systematically examine microscopic interactions of benzothiadiazine with the cell membrane at liquid-crystalline phase conditions. From data obtained through radial distribution functions, hydrogen-bonding lengths, and potentials of mean force based on reversible work calculations, we have observed that benzothiadiazine has a strong affinity to stay at the cell membrane interface although it can be fully solvated by water in short periods of time. Furthermore, benzothiadiazine is able to bind lipids and cholesterol chains by means of single and double hydrogen-bonds of different characteristic lengths.