Innovative Biosystems and Bioengineering

Effects of Acute Alcohol Intoxication on Testicular Dna Stability, Gene Expression of Cytochromes CYP3A and CYP2E1, and Serum Pool of Free Amino Acids in Rats

Larysa Bondarenko — 2024-12-09

Background. Alcohol's toxic effects on the organism is a long-known medical problem. Alcohol's damaging effect is the end result of the complex interplay between ethanol metabolism, inflammation and innate immunity. Previously, we studied the long-term consequences of chronic alcoholism and demonstrated that especially profound changes were in testes on the level of proteome and genome.

Objective. This work aimed to study short-term acute alcohol intoxication (AAI) effects for rat testis DNA fragmentation, cytochromes CYP3A and CYP2E1 genes expression, and serum pool of free amino acids in rats.

Methods. Wistar albino male rats were divided into 2 groups (8 animals in each group): 1 – Control (intact rats), and 2 – AAI (rats with short-term acute alcohol intoxication). AAI was induced by repeated administration per os 40% ethanol solution in a dose 7 ml/kg body weight, for 7 days. Contents of amino acids in serum, testes mRNA CYP2E1 and CYP3A2 expression, and DNA fragmentation were evaluated.

Results. In our experiments, the development of acute alcohol intoxication (AAI) led to increased DNA fragmentation processes in the testes of adult rats compared to the control group. Additionally, in the serum of ethanol-treated rats, the levels of histidine increased by 1.67 times and glutamine by 1.13 times in correlation with this pathology. Conversely, the levels of valine, phenylalanine, as well as non-essential and essential amino acids, decreased. Furthermore, there was a statistically significant increase in the expression of CYP2E1 and CYP3A2 genes in rat testes under the conditions of AAI.

Conclusions. In conclusion, investigation of rats' short-term alcohol administration effects permitted us to obtain the picture of complex metabolomic changes at the different levels. The main outcome of rats short-term ethanol administration in our experiments seems to be to some extent similar to changes described for rats with chronic alcohol consumption. Our results demonstrated profound changes in testes affecting the state of the genome, transcription processes and the exchange of amino acids and proteins. We suggest that the revealed testicular metabolic disorders could have negative implications on cellular regulation of spermatogenesis even under short-term ethanol exposure.

Effect of Fullerene C60 on Tomato Plants

Anastasiia Buziashvili — 2024-12-09

Background. Fullerenes, one of the allotropic forms of carbon, are the focus of intensive research in nanobiotechnology. Due to their unique physicochemical properties, there is growing interest in using them to enhance plant productivity and provide protection against various stresses. However, data on the effects of fullerenes on different plants are often contradictory.

Objective. To investigate the effect of colloidal water-soluble fullerene C₆₀ on various stages of tomato development.

Methods. The study examined the influence of fullerene C₆₀ (0–50 µg/ml) on the seed germination of the Money Maker cv. and the growth and development of seedlings and adult plants. For this purpose, morphophysiological parameters such as shoot and root length, number of lateral and adventitious roots, fresh weight, accumulation of photosynthetic pigments, and nitric oxide (II) were assessed.

Results. Both positive and some phytotoxic effects of fullerene C₆₀ on tomatoes were observed. Specifically, inhibition of seedling growth and a decrease in chlorophyll b and carotenoid content in adult plants were noted. Positive effects included an increased seed germination rate, higher fresh weight, greater length and number of adventitious roots in tomatoes grown in the presence of fullerene C₆₀, and a higher nitric oxide (II) content in adult plants, which may contribute to enhanced stress resistance.

Conclusions. Fullerene C₆₀ at a concentration of 25 μg/ml has a predominantly positive effect on tomato development and may be considered a promising nanomaterial for plant biotechnology.

Design and Development of Novel Herbal Suppository Formulation for Prostatitis Treatment

Olena Golembiovska — 2024-12-09

Background. Prostatitis is a common inflammatory condition of the prostate gland, often treated with antibiotics and anti-inflammatory drugs, which may have limited efficacy and side effects. Herbal therapies offer a promising alternative due to their natural anti-inflammatory properties.

Objective. This study aims to design, develop, and evaluate physicochemical, biopharmaceutical and microbiological properties of novel herbal suppository formulations for the treatment of prostatitis, optimizing their biopharmaceutical profiles.

Methods. Excipients were selected to form suppositories with herbal ingredients known for their anti-inflammatory effects, specifically lovage, saw palmetto, and calendula CO₂ extracts. The formulations were assessed based on key quality parameters, including organoleptic characteristics, average mass, melting point, deformation time, disintegration time, microbiological purity, and pH, ensuring compliance with European Pharmacopoeia (Ph. Eur.) standards. Biopharmaceutical studies compared the release profiles of active compounds from different suppository bases.

Results. Suppositories based on Witepsol® H15 exhibited satisfactory organoleptic properties, appropriate melting points, and acceptable deformation and disintegration times. The pH levels were within the required range, and microbiological tests confirmed purity. Biopharmaceutical evaluations showed that Witepsol® H15 suppositories had the most appropriate release rates of active compounds compared to those made with other bases.

Conclusions. The novel herbal suppositories developed in this study show promise for managing prostatitis based on preliminary evaluations of their physicochemical, microbiological, and biopharmaceutical properties. Further clinical studies are required to confirm their efficacy and safety.

Effect of Acids on Biomass Growth and Development of Chlorella vulgaris Culture

Alona Vdovychenko — 2024-12-11

Background. Influence of dissolved nitrogen and sulfur oxides, as components of flue gases, on the cultivation of microalgae Chlorella vulgaris.

Objective. To study biomass production and changes in cells of the microalgae Chlorella vulgaris with the introduction of nitrogen and sulfate acids tor simulate the effects of dissolved nitrogen and sulfur oxides, aiming to develop biotechnology for the utilization of gas emissions by microalgae.

Methods. The effect of the constant introduction of small concentrations of nitrogen (up to 0.47%) and sulfate (up to 1.5%) acids on the development of Chlorella vulgaris cultures, pH changes in the cultural environment, and biomass growth were studied.

Results. The utilization of gaseous emissions by Chlorella vulgaris depends on the content of nitrogen and sulfur oxides that are constantly supplied to the cultivation medium, the initial biomass concentration, and pH. It was determined that for an initial cells concentration of (85 ± 5)×10⁴ cells/ml in the culture medium, the threshold values of acids that do not cause significant changes in the cultivation process are 0.1% H₂SO₄ and 0.19% HNO₃. At a sulfuric acid concentration of 0.2%, the culture cells become discolored.

Conclusions. It is shown that with a constant supply of sulfuric or nitric acids above the threshold values of 0.1% H₂SO₄ and 0.19% HNO₃, the pH of the culture medium decreases, leading to the suspension of development and the death of the Chlorella vulgaris culture. Therefore, controling these parameters will improve the ecological state of the environment and form the basis for developing biotechnology for the utilization of gas emissions by microalgae.

Cytodestructive Effects of Photodynamic Exposure in Primary Cultures of Malignant Glioma Cells

Volodymyr Rozumenko — 2024-12-16

Background. Photodynamic therapy (PDT) is a promising adjuvant method for the treatment of malignant gliomas (MG), including tumors with continued growth and tumor recurrences. For the clinical application of PDT, it is important to substantiate the effectiveness of the cytodestructive effect of the combined use of laser irradiation (LI) and photosensitizer (PS).

Objective. To evaluate the cytodestructive effects of photodynamic exposure with the use of PS chlorine E6 on primary MG cell cultures.

Methods. Primary cell cultures were obtained from samples of biopsy material from patients (n = 6) with a verified diagnosis: 3 primary tumors (1 case of diffuse astrocytoma, NOS (G3), 1 – glioblastoma (GB), NOS (G4), 1 – gliosarcoma (G4)) and 3 – with continued tumor growth (1 – diffuse astrocytoma, NOS (G3), 1 – oligodendroglioma, NOS (G3) and 1 – GB, NOS (G4). Groups of cell cultures included: 1) control – cultured in a standard nutrient medium and experimental; 2) cultured with the addition of chlorine E6 (2.0 mg/ml); 3) cultivated without the addition of PS and subjected to LI; 4) cultivated with the addition of chlorine E6 and subsequent exposure to LI. After 24 h, morphological and morphometric studies were carried out.

Results. The primary MG cultures were characterized by different growth dynamics; mitotic activity of tumor cells varied from the highest rate in the culture of primary GB to lower values – in cultures of recurrent GB and primary astrocytoma and gliosarcoma, and the lowest – in cultures of continued growth of astrocytoma and oligodendroglioma after combined treatment. Direct exposure to chlorine E6 and LI reduced the total number of cells in the culture and their mitotic activity. The greatest cytodestructive effect was achieved with the combined effect of chlorine E6 and LI: the effective dose in the case of primary astrocytoma cells is 10 J/cm² in pulse mode; for cells of primary GB and gliosarcoma, recurrent astrocytoma and oligodendroglioma, the effective dose is 25 J/cm² in pulsed mode. In the case of GB cells, continued growth, a dose of 25 J/cm² is effective for both continuous and pulsed modes of LI.

Conclusions. Primary cell cultures of MG obtained directly from tumor tissue are an adequate model for evaluating the effectiveness of the cytodestructive effect of the combined use of LI and PS for PDT.