Vol 26, No 11 (2023)

Relevance. Both highly active pharmaceutical substances and excipients can be target products for the complex processing of medicinal plant raw materials. For example, naphthoquinones are used as antimicrobal, antifunginal and polysaccharides are used as fillers, thickeners, and sorbents. Fruits, roots, and waste from the wood processing industry serve as raw material sources of polysaccharides. The development of low-waste technologies for processing medicinal plant raw materials is a current area of research.

The purpose of the work is to substantiate the possibility of complex processing of black walnut pericarp with the production of biologically active and auxiliary substances using low-waste technology based on phytochemical research.

Material and methods. The object of study is the pericarp of Juglans nigra L. (Juglandaceae). The raw materials were collected in October 2021 in a park in Pyatigorsk. It was dried and crushed into powder. Naphthoquinones (juglone and its derivatives) were detected by thin layer chromatography. The content of naphthoquinones was determined by spectrophotometry, extractives released by chloroform, polysaccharides by gravimetry, the average molecular weight of water-soluble polysaccharides by viscometry, the surface activity of water-soluble polysaccharides by stalagmometry, and the distribution coefficient of polysaccharides in the diethyl ether-water system by conductometry.

Results. The possibility of obtaining two products in one technological process is justified by the results of experiments: 1) active pharmaceutical substance (yield 1.5%) for obtaining external forms (ointments, creams) with antifungal, antiviral activity, containing naphthoquinones (naphthoquinones in terms of juglone 12.41±0.3%); 2) excipients polysaccharides (yield 8.8%), whose molecules have a thread-like structure, which explains their good solubility in water, surface activity and polyelectrolyte properties.

Conclusions. As a result of a phytochemical study of the pericarp of Juglans nigra L., it was established that both a substance containing polyphenolic compounds and polysaccharides suitable as auxiliary substances can be obtained in the process of complex processing.

Relevance. Natural mineral clays are widely used not only in technology, but also in medicine, cosmetology, and the food industry, due to their unique properties. The high degree of dispersion, sorption-adhesive properties of clay materials, as well as the rich mineral composition also determine their therapeutic effect.

Depending on the area and purpose of use, clays are subjected to heat treatment, which can lead to a change in their properties.

The purpose of the work is to study the influence of various heat treatment modes on the sorption properties of clay for its further use in pharmaceutical practice.

Material and methods. The object of study is Kimmeridgian blue powdery of clay blue Undory in the Ulyanovsk region. Determination of clay moisture content was carried out using an AND MF–5 moisture analyzer (Japan). To completely remove moisture from clay by high-temperature treatment, an SNOL 7.2/1100 electric furnace (Lithuania) was used. The influence of the clay calcination regime on the thermal stability of the clay powder and the ongoing endo- and exothermic effects with loss of sample mass was studied using a Q-1500D derivatograph (Hungary). Adsorption activity was assessed by direct spectrophotometry using an SF-56 “LOMO-Spectrum” spectrophotometer (Russia).

Results. DTA of the derivatogram showed the thermal stability of the clay powder in the temperature range from 20°C to 900°C with variations in endo- and exothermic effects. Based on these data, heat treatment modes and sample exposure time were selected. The experiment showed a statistically significant decrease in adsorption activity with increasing temperature and various variations in exposure time.

Conclusions. To preserve the adsorption properties of clay blue Undory when removing moisture from it, it is not rational to use high-temperature treatment. The most optimal parameters for drying clay are: temperature 80 ° C, exposure time - 240 minutes.

Relevance. Oxidative stress is an imbalance between the production of reactive oxygen species and the ability of the endogenous antioxidant system, and is also part of the pathogenesis of a number of diseases. Recently, antioxidants have been included in the complex treatment of many pathological conditions, so the search for compounds with an antioxidant effect is relevant.

Aim. Study of the antioxidant activity of 2-aryl-7,7-dimethyl-5-oxo-5,6,7,8-tetrahydroquinoline-4-carboxylic acids in a model of oxidative stress induced by hydrogen peroxide in a cell culture of E. coli strain "Ecolume."

Material and methods. 2-Aryl-7,7-dimethyl-5-oxo-5,6,7,8-tetrahydroquinoline-4-carboxylic acids were obtained as a result of the interaction of 4-aroyl-2,4-dioxobutane acids with 3-amino-5,5-dimethylcyclohex-2-enone. The evaluation of the antioxidant activity of the synthesized compounds was carried out by an indirect method of assessing changes in the fluorescence of E. coli cell culture strain "Ecolume" using a model of oxidative stress induced by hydrogen peroxide.

Results. Among 2-aryl-7,7-dimethyl-5-oxo-5,6,7,8-tetrahydroquinoline-4-carboxylic acids, compounds with antioxidant activity exceeding the reference standard – trolox were found.

Conclusions. Investigation of the antioxidant activity of 9 compounds of the 2-aryl-7,7-dimethyl-5-oxo-5,6,7,8-tetrahydroquinoline-4-carboxylic acid series using E. coli cell culture as a biosensor.

Relevance. Plant-derived compounds are attractive alternative to antibiotic growth promoters in livestock and poultry production. In our previous study, dietary supplementation of broiler chickens with the flavonoid quercetin, the phenolic aldehyde vanillin, and the o-hydroxycinnamic acid lactone umbelliferon (7-hydroxycoumarin) modulated the ceca microbiome, reduced systemic inflammation, and increased production efficiency, as measured by the EPEF index.

Objectives. To evaluate the content of free and conjugated (glucuronidated) quercetin, vanillin and umbelliferon in the muscles and liver of broiler chickens fed a 35-day basic diet supplemented with these compounds.

Material and methods. Quercetin, vanillin and umbelliferon were extracted with acetonitrile and formic acid, evaporated and re-dissolved in deionized water, and then purified using HLB cartridges. To release the conjugated compounds, hydrolysis with glucuronidase/arylsulfatase was additionally performed. The resulting samples were analyzed by HPLC-MS using a LCMS-8050 gas chromatograph - mass spectrometer (Shimadzu Corporation, Japan) with LabSolutions software installed.

Results. Tissue samples from broiler chickens fed quercetin did not contain this compound in a detectable concentration range, which may be a consequence of the effective quercetin metabolism by intestinal microflora and xenobiotic inactivation systems. Surprisingly, quantification of vanillin showed its presence in all analyzed samples, regardless of presence or absence in supplements. This result cannot be interpreted unequivocally and is currently speculated to be due to vanillin or similar cross-reacting substances in the basic diet. Umbelliferon concentrations of 17 ng/g (free) and 125 ng/g (sum of free and conjugated forms) were detected in only one liver sample of broiler chickens that received this compound at the maximum dosage (3.0 mg per 1 kg of feed).

Conclusion. The absence or low accumulation of free and glucuronidated quercetin, vanillin and umbelliferon in the muscles and liver of diet-supplemented broiler chickens has been shown. This result indicates the safety of these plant-derived molecules in poultry production without possible migration through food chains.

Relevance. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test is one of the most common used methods for determining cell viability (cytotoxicity) during cell response to plant’s extract or individual extract’s compound. Despite of extensive use of this method it remains unclear whether plasma membrane of cells is destroyed by MTT-formazan crystals during MTT reduction to MTT-formazan; whether the MTT-formazan is lost after cell destruction by MTT-formazan crystals.

The purpose of this study was to assess by microscopy and spectrophotometry methods the possibility of MTT-formazan releasing into cultural medium during MTT reduction to MTT-formazan by HeLa cells.

Material and methods. HeLa cells were grown on coverslips in Weighing bottles, incubated with MTT (0.1 or 0.5 mg/ml final concentrations) and visualized by light microscopy after 30 min, 60 min, 120 min and 240 min after MTT addition. For spectrophotometry measurements cells (5×10⁴ cells/well) were added to a 96-well plate followed by MTT. The absorbance spectrum was recorded after 30 min, 60 min, 120 min and 240 min after MTT addition.

Results. Microscopic examination revealed two phases in MTT reduction by cells to MTT-formazan – the formation of granules and crystals of MTT-formazan. The maximum ratio of crystals to granules of MTT-formazan and almost complete destruction of cells were observed by light microscopy at 240 min of incubation with MTT. However, in the study by spectrophotometry after cell destruction by dimethyl sulfoxide the values of optical density of the MTT-formazan after 120 min as well as 240 min of incubation were almost the same. Therefore, the transformation of MTT-formazan from granules to crystals does not affect the optical density despite of cell destruction. Culture medium without cells did not reduce MTT to MTT-formazan. In the supernatant from cells without their destruction the significant production of MTT-formazan was also not observed.

Conclusions. MTT-formazan crystals do not dissolve in the culture medium but dissolve in hydrophobic solvents such as dimethyl sulfoxide. In this regard the destruction of cells by MTT-formazan crystals does not lead to significant loss of MTT-formazan during MTT reduction by HeLa cells and does not significantly effect on results of cytotoxicity determined by MTT test.

Relevance. The role of nerve growth factor (NGF) deficiency in the pathogenesis of Alzheimer's disease (AD) is well-known. The clinical application of full-size neurotrophin is limited due to its low bioavailability and the risk of adverse effects. At the V.V. Zakusov Institute of Pharmacology, a dimeric dipeptide mimetic of the 4th loop of NGF, compound GK-2 (hexamethylenediamide bis-(N-monosuccinyl-L-glutamyl-L-lysine), was created. It selectively activates specific TrkA receptors and possesses neuroprotective and neuroregenerative properties. Additionally, GK-2 lacks the main side effects of NGF, namely hyperalgesia and weight loss.

The aim of the current study was to investigate the influence of GK-2 on rat memory under the conditions of the scopolamine-induced model of AD.

Material and Methods. Scopolamine was administered intraperitoneally to rats at a dose of 2 mg/kg for 32 days. Simultaneously with scopolamine, animals were intraperitoneally injected with GK-2 at doses of 0.5 and 1 mg/kg. Following the administration of the compounds, a novel object recognition test was conducted to assess both short-term and long-term memory.

Results. Rats receiving scopolamine exhibited a statistically significant decline in long-term memory. The dipeptide GK-2 at a dose of 1 mg/kg completely counteracted the development of this impairment.

Conclusion. The dipeptide mimetic of nerve growth factor, GK-2, shows promise for further investigation as a potential therapeutic agent for the treatment of AD.

Alzheimer's disease (AD) is the most common form of dementia that primarily affects older adults and most often begins with memory loss followed by progressive impairment of behavioral and cognitive functions. Despite the fact that the main pathological signs of AD are considered to be extracellular deposits of beta-amyloid in the form of amyloid plaques and intracellular accumulation of hyperphosphorylated tau protein in the form of neurofibrillary tangles, recently more and more attention at the cellular and molecular levels has been paid to other important processes accompanying development of the disease. In modern research of neurodegenerative diseases, the role of mitochondria is receiving increasing interest. The mitochondrial cascade hypothesis suggests that mitochondrial dysfunction plays a key role in the progression of these neurodegenerative processes. Recent research shows that cells have the ability to exchange mitochondria among themselves. This process, known as horizontal mitochondrial transfer, allows cells to exchange both healthy and damaged or dysfunctional mitochondria, moving them from one cell to another for further repair or degradation, which raises the possibility of using mitochondrial transplantation as a therapy for neurodegenerative diseases.

In this article, we consider two aspects: horizontal mitochondrial transfer and mitochondrial transplantation. Horizontal mitochondrial transfer opens new horizons in understanding cellular communication and interactions. The methods of horizontal transfer of mitochondria discussed in the article are presented and described in detail. Additionally, we review the relevance and innovative nature of mitochondrial transplantation, a procedure in which healthy mitochondria are transferred to cells or organs with dysfunctional mitochondria. We will discuss various mitochondrial transplantation methods and their potential applications in medicine. The article will provide information on new research and perspectives in the field of mitochondrial biology and therapeutics, expanding the understanding of the function and role of mitochondria in living organisms.