Study of hepatotoxins influence in vitro on basic biochemical indicators of liver functional state

An antimicrobial drug of the fluoroquinolone group, ciprofloxacin, is widely used in Ukraine. However, some researchers have noted the probable hepatotoxicity of this drug with prolonged use or use of high doses of ciprofloxacin. The aim of this study was to compare the effects of carbon tetrachloride, as a classical model of hepatocyte injury, with the effects of ciprofloxacin. The aim of the study was to investigate the biochemical parameters of the liver when simulating toxic damage to hepatocytes with carbon tetrachloride or ciprofloxacin. Materials and methods. The study was carried out on isolated rat hepatocytes, in whose culture medium carbon tetrachloride or ciprofloxacin was added. After incubation in the supernatant and cell homogenate, the activities of marker enzymes of cytolysis were determined: ALT, AST, γ-GTP, LF, LDH, DC and MDA. Results. The introduction of ciprofloxacin into the culture of hepatocytes at a concentration of LC50 caused changes in biochemical parameters similar to those caused by carbon tetrachloride. Thus, an increase in ALT, AST, γ-GTP, LF, LDH, DC and MDA was observed when CCl4 or ciprofloxacin was added to the culture. Conclusion. Incubation of rat hepatocytes with carbon tetrachloride or ciprofloxacin caused an increase in the level of enzymes and lipid peroxidation products. These parameters are indicators of gross changes in cells, which are the result of impaired keto acid formation, impaired redox reactions, impaired glycogen production

Morphofunctional features of proliferating cells exposed to PSMA peptide

The effect of the synthetic PSMA peptide on dividing cells of laboratory animals was studied. The experiment was carried out on male white laboratory mice of the BALB/c-line. The toxic effect of PSMA peptidi was evaluated at therapeutic (1.4 μg / kg of animal weight or 0.04 μg / animal) and subtoxic (140 μg / kg of animal weight or 4.0 μg / animal) doses. The cytotoxic effect of PSMA peptide on red bone marrow cells and cambial intestinal cells of the of laboratory mice was determined. A decrease in the proliferative activity of the colon crypt cells was revealed upon administration of a subtoxic dose of the PSMA peptide and there were no signs of toxic damage to the red bone marrow cells of animals. Key words: toxicity, proliferation, synthetic peptides, mitotic index, micronucleus test.

The Philosophical and Evidentiary Basis of Homoeopathic Immunisation: a Response to Teixeira

Homeopathic immunisation was first used in 1798, but remains controversial with some homeopathic practitioners. Teixeira supported the use of genus epidemicus (GE) remedies, but strongly condemned the use of nosodes for disease prevention. However Teixeira failed to fully understand the Principal of Similars, he used a double standard when comparing evidence using GE remedies and nosodes, he misread information demonstrating the safety of long-term HP, and he appeared to be unaware of scientific evidence which is available supporting the prophylactic use of nosodes. These four areas are addressed in turn, and evidence from 1798 to 2012 is presented showing that appropriate homoeopathic immunisation using GE remedies and/or nosodes has the potential to prevent much suffering without any risk of possible short-term toxic damage or long-term energetic adverse effects.

Histomorphometric study of rat liver during the treatment of the acute toxic injury

Introduction. There are a few effective therapies are available for acute liver injury at present. The aim of the study was to investigate histological and morphometric changes in the liver using models of toxic damage caused by carbon tetrachloride (CCl4 ) and acetaminophen during correction with Oxymethyluracil (OMU). Material and methods. A total of ninety rats were divided into 18 groups. The treatment of acute liver damage models caused by a single injection of CCl4 or acetaminophen was carried out using “Heptor”, “Mexidol”, and OMU. The correction was carried out twice (sacrificed 24 hours after intoxication) and four times (sacrificed 72 hours after intoxication). Liver tissues were processed using standard histological techniques (H&E). A semi-quantitative assessment was performed using a scale based on the severity of liver cell deaths. Results. Twenty-four hours after administration of CCl4 or 72 hours after administration of acetaminophen, the treatment with OMU led to a decrease in liver cell death compared to the group with administration of only CCl4 or acetaminophen. Seventy-two hours after CCl4 and 24 hours after acetaminophen intoxication, these groups with the OMU treatment did not differ from those of the carbon tetrachloride- or acetaminophen-induced liver injury groups, respectively. Conclusion. Thus, on the model of CCl4 liver injury, the treatment with OMU is more effective for 24 hours. In the case of acetaminophen intoxication, the effectiveness of treatment with OMU is better for 72 hours. The results obtained are possibly associated with a different mechanism of the damaging effect of the studied toxicants.

Formaldehyde induces ferritinophagy to damage hippocampal neuronal cells

Formaldehyde (FA) causes neurotoxicity and contributes to the occurrence of neurodegenerative diseases. However, the mechanism of FA-induced neurotoxicity has not been fully elucidated. Ferritinophagy, an autophagy process of ferritin mediated by the nuclear receptor coactivator 4 (NCOA4), is a potential mechanism of neurotoxicity. In this study, we explored whether ferritinophagy is associated with the neurotoxicity of FA. Our results showed that FA (50, 100, 200 μM; 24 h) exposure upregulated ferritinophagy in the mouse hippocampal neuronal HT22 cells, which was evidenced by the upregulated autophagic flux, the increased colocalizations of NCOA4 with ferritin heavy chain (FTH1) and NCOA4 with microtubule-associated protein 1 light chain-3B (LC3B), the augmented expression of NCOA4, and the reduced content of FTH1. We also found that FA (0.1, 1, and 10 μmol, i.c.v., 7d) administration boosted ferritinophagy in the hippocampus of Sprague-Dawley (SD) rats, which was demonstrated by the accumulated autophagosomes, the increased expressions of LC3II/I and NCOA4, and the decreased contents of p62 and FTH1 in the hippocampus. Further, we confirmed that inhibition of ferritinophagy by silencing the expression of NCOA4 decreased FA-induced toxic damage in HT22 cells. These results indicated that FA induces neurotoxicity by promoting ferritinophagy. Our findings suggest a potential mechanism insight into the FA-induced neurotoxicity, which in turn provides a new thought for the treatment of FA-related neurodegenerative diseases.

Determination of the activity of pyruvate kinase isoforms in normal conditions, in toxic liver damage and during the process of its regeneration

Objective. To evaluate the activity of pyruvate kinase (PK) isoforms in normal conditions, in toxic damage of the liver and during its regeneration.Materials and methods. An experimental study was carried out on 45 Wistar rats. Toxic liver damage was induced by the intraperitoneal administration of carbon tetrachloride. Mechanical damage was simulated by the surgical resection of the liver. The levels of PK isoforms R/L and M in the blood serum and liver tissue of the laboratory animals were measured with an ELISA test.Results. It has been found that the level of PK isoform M signifcantly increases in chronic toxic liver damage, which may indicate the activation of the processes of liver cell proliferation in response to the damaging effect of hepatotoxin (Mann-Whitney U Test: Z = 2.143; p = 0.032). After liver resection, the level of PK R/L, which characterizes the activation of glycolysis, increased and the level of pyruvate kinase M increased signifcantly, which reflected the processes of reparative regeneration in the liver.Conclusion. The serum level of PK isoforms may be used as a laboratory criterion for the activity of reparative regeneration processes, which can be used to evaluate the reparative potential of the liver in case of toxic or mechanical damage, as well as in chronic diffuse diseases.

Toxic Damage to Motor Neurons

Abstract—Amyotrophic lateral sclerosis (ALS) is a multifactor disease in the development of which both genetic and environmental factors play a role. Specifically, the effects of organic and inorganic toxic substances can result in an increased risk of ALS development and the acceleration of disease progression. It was described that some toxins can induce potentially curable ALS-like syndromes. In this case, the specific treatment for the prevention of the effects of the toxic factor may result in positive clinical dynamics. In this article, we review the main types of toxins that can damage motor neurons in the brain and spinal cord leading to the development of the clinical manifestation of ALS, briefly present historical data on studies on the role of toxic substances, and describe the main mechanisms of the pathogenesis of motor neuron disease associated with their action.

Effect of combined transplantation of multipotent mesenchymal stromal cells and stellate liver cells on the morpho-functional state of the liver after adminitration of CCL4

Цель исследования - изучение влияния сочетанной трансплантации мультипотентных мезенхимальных стромальных и звездчатых клеток печени на морфофункциональное состояние печени после ее токсического повреждения, вызванного введением CCl4. Задачи исследования: 1. Оценка морфометрических показателей печени после сочетанного введения мультипотентных мезенхимальных стромальных (ММСК) и звездчатых клеток печени (ЗКП) при токсическом повреждении печени CCl4. 2. Изучение влияния котрансплантации ММСК и ЗКП на биохимические показатели крови при токсическом повреждении печени CCl4. 3. Исследование активности системы репарации ДНК после введения ММСК и ЗКП при токсическом повреждении печени CCl4. Методика. Эксперименты выполнены на 63 белых лабораторных мышах-самцах в возрасте 7-8 мес. Токсический гепатит воспроизводили у всех животных внутрибрюшинным введением CCl4 (50 мкл/мышь), затем из них формировали опытную и контрольную группы. Животным опытной группы в латеральную хвостовую вену вводили суспендированные в 0,2 мл 0,9 % раствора NaCl ММСК, полученные из хориона плаценты мышей-самок (4 млн клеток/кг, 120 тыс клеток/мышь), и ЗКП - 9 млн клеток/кг (270 тыс клеток/мышь). Животным контрольной группы вводили в латеральную хвостовую вену 0,2 мл 0,9 % раствора NaCl. Внутривенные инъекции осуществляли однократно через 1 ч после введения CCl4. Использовлили ММСК 3-го пассажа. Трансплантированные ЗКП не подвергались культивированию. Исследовали влияние сочетанной трансплантации ММСК и ЗКП на морфофункциональное состояние печени на 1-е, 3-и, 7-е сут после введения CCl4. Результаты. Сочетанная трансплантация мультипотентных мезенхимальных стромальных и звездчатых клеток печени при токсическом поражении печени приводит к повышению митотической активности гепатоцитов, увеличению числа двуядерных гепатоцитов, повышению ядерно-цитоплазматического отношения. Введение стволовых клеток способствует снижению запрограммированной клеточной гибели гепатоцитов за счет повышения активности ферментов репарации семейства PARP. Заключение. Сочетанная трансплантации ММСК и ЗКП положительно влияет на морфофункциональное состояние печени в условиях ее токсического повреждения. Значимым механизмом восстановления морфофункционального состояния печени можно считать влияние трансплантируемых ММСК и ЗКП на систему репарации клеток. The aim of this study was to investigate the effect of combined transplantation of multipotent mesenchymal stromal and stellate liver cells on the morphofunctional state of the liver after toxic damage caused by carbon tetrachloride. Objectives of the study: 1. To evaluate changes in liver morphometric parameters after combined administration of multipotent mesenchymal stromal cells (MMSC) and hepatic stellate cells (HSC) in toxic liver damage. 2. To study the effect of cotransplantation of MMSC and HSC on changes in blood biochemical parameters in toxic liver damage. 3. To investigate the activity of the DNA repair system after the introduction of MMSC and HSC in toxic liver damage. Methods. The experiments were performed on 63 white laboratory male mice aged 7-8 mos. Toxic hepatitis was caused by intraperitoneal administration of carbon tetrachloride (CCl4) at a dose of 50 µl/mouse. The mice were divided into experimental and control groups. Animals of the experimental group were injected into the lateral caudal vein with MMSCs obtained from the chorion of the placenta of female mice and with HCP at doses of 4 million cells/kg (120 thousand cells/mouse) and 9 million cells/kg (270 thousand cells/mouse), respectively, suspended in 0.2 ml of 0.9% NaCl solution. Animals of the control group were injected with 0.2 ml 0.9 % NaCl into the lateral caudal vein. Intravenous injections were performed 1 hr after the administration of carbon tetrachloride. Rats were administered with MMSCs of the third passage. Transplanted HSC had not been subjected to cultivation. The effect of combined MMSC and HSC transplantation on the morpho-functional state of the liver was studied on the 1st, 3rd, and 7th days after administration of carbon tetrachloride. Results. The combined transplantation of multipotent mesenchymal stromal and hepatic stellate cells leads to an increase in the mitotic activity of hepatocytes, an increase in the number of binuclear hepatocytes, and an increase in the nuclear-cytoplasmic ratio. Administration of stem cells helps to reduce the programmed cell death of hepatocytes by increasing the activity of repair enzymes of the PARP family. Conclusion. Combined transplantation of MMSC and hepatic stellate cells (HSC) has a positive effect on the morphofunctional state of the liver in conditions of its toxic damage. A significant mechanism for the restoration of the morphological and functional state liver, the effect of transplanted MMSC and HSC on the cell repair system can be considered.

MINERAL HOMEOSTASIS OF THE ORAL FLUID IN CHILDREN AND ADOLESCENTS WHO HAVE UNDERGONE TREATMENT OF MALIGNANT NEOPLASMS

Annotation. The authors' study highlights the urgent problem of the development of toxic damage to the dentition under the influence of anticancer therapy in children and adolescents. The aim of the study was to study the violation of the homeostasis of the oral fluid and the severity of the carious process. Materials and methods. As part of a pilot study at the Russian Field Treatment and Rehabilitation Center, 63 children were studied the severity of mineral homeostasis disorders and damage to the dentoalveolar system. Results. In 67% of patients in the main group, there was an increased tendency to form carious cavities. 54.5% of them have complicated caries. One third of patients had more than five carious teeth. In the control group, carious lesions of the teeth in only three patients (20%), with the involvement of 1-3 teeth in the pathological process. Violation of enamel formation was diagnosed in 36% of children of the main group. There is a tendency for the accumulation of the studied microelements in the oral fluid, while in the blood serum there is a normal or insignificant decrease in their content. According to the literature, this may indicate destruction in the oral cavity. Deviations in the homeostasis of chemical elements in children who completed the treatment of malignant neoplasms and children in the control group were revealed. An increased content of osteotrophic microelements was found to correlate with the destructive processes of the teeth. Conclusion. It seems appropriate to study the severity of morphological changes in hard tissues of teeth and the electrolyte composition of mixed saliva in children and adolescents who have been cured of malignant neoplasms. The results of studies of gastric cancer, as an indicator of metabolic disorders, will make it possible not only to reveal the imbalance of macro- and microelements, but also to establish the effectiveness of adaptation mechanisms aimed at normalizing the elemental composition of the oral fluid.