Serum Of Experimental Animals Research Articles

Naringenin Impedes the Differentiation of Mouse Hematopoietic Stem Cells Derived from Bone Marrow into Mature Dendritic Cells, thereby Prolonging Allograft Survival.

The use of immature dendritic cells (imDCs) to induce donor-specific immunotolerance following in vivo stimulation is limited by their low rate of induction and their tendency to undergo maturation. We derived imDCs from bone marrow hematopoietic stem cells (HSCs-imDCs). We then tested the ability of naringenin (Nar) to impede the maturation of HSCs-imDCs for inducing transplantation immune tolerance. HSCs derived from bone marrow were collected and induced to differentiate into imDCs by treating with Nar (Nar-HSCs-imDCs). Flow cytometry was used to evaluate DC surface markers, apoptosis, and endocytic ability. The ability of DCs to influence the in vitro proliferation of T cells and of regulatory T cells (Tregs) was analyzed by mixed lymphocyte reaction assays. Enzyme-linked immunoassays were used to quantify cytokine levels in supernatants from co-cultured DCs and Tregs, as well as in the serum of experimental animals. The level of immunotolerance induced by Nar-HSCs-imDCs was evaluated by skin grafting in recipient Balb/c mice, while the Kaplan-Meier method was used to statistically evaluate graft survival. Compared with HSC-imDCs, Nar-HSCs-imDCs showed higher expression of cluster of differentiation 11c (CD11c), but lower expression levels of CD80, CD86, and major histocompatibility complex class II. Nar-HSCs-imDCs also showed stronger inhibition of T cells and higher Treg cell proliferation. Interleukin 2 (IL-2) and interferon gamma levels were downregulated in Nar-HSCs-imDCs, whereas IL-4, IL-10, and transforming growth factor beta levels were upregulated. The rate of apoptosis and endocytic capacity of Nar-HSCs-DCs increased significantly after treatment with lipopolysaccharide. HSCs-imDCs or Nar-HSCs-imDCs were injected into Balb/c mice via the tail vein 7 days before skin grafting. Significantly reduced donor-specific CD4+ T cells and induced proliferation of CD4+CD25+FoxP3+ Treg cells were observed in the spleen of mice from the Nar-HSCs-imDCs group, especially at a dose of 106 Nar-HSCs-imDCs. The latter group also showed significantly prolonged survival of skin grafts. Nar-HSCs-imDCs markedly improved the acceptance of organ allografts, offering a potentially new strategy for inducing immune tolerance in transplantation.

Synaptamide Modulates Astroglial Activity in Mild Traumatic Brain Injury

At present, the study of the neurotropic activity of polyunsaturated fatty acid ethanolamides (N-acylethanolamines) is becoming increasingly important. N-docosahexaenoylethanolamine (synaptamide, DHEA) is a highly active metabolite of docosahexaenoic acid (DHA) with neuroprotective, synaptogenic, neuritogenic, and anti-inflammatory properties in the nervous system. Synaptamide tested in the present study was obtained using a chemical modification of DHA isolated from squid Berryteuthis magister liver. The results of this study demonstrate the effects of synaptamide on the astroglial response to injury in the acute (1 day) and chronic (7 days) phases of mild traumatic brain injury (mTBI) development. HPLC-MS study revealed several times increase of synaptamide concentration in the cerebral cortex and serum of experimental animals after subcutaneous administration (10 mg/kg/day). Using immunohistochemistry, it was shown that synaptamide regulates the activation of GFAP- and S100β-positive astroglia, reduce nNOS-positive immunostaining, and stimulates the secretion of neurotrophin BDNF. Dynamics of superoxide dismutase production in synaptamide treatment confirm the antioxidant efficacy of the test compound. We found a decrease in TBI biomarkers such as GFAP, S100β, and IL-6 in the blood serum of synaptamide-treated experimental animals using Western blot analysis. The results indicate the high therapeutic potential of synaptamide in reducing the severity of the brain damage consequences.

Зміни деяких біохімічних показників сироватки крові у разі дво- та шеститижневого застосування терапевтичних доз опіоїду та на ранніх термінах відміни препарату

An important issue today is the safety of opioids for the treatment of chronic pain syndrome, which accompanies a number of somatic diseases. Most drugs or their metabolites are excreted through the kidneys and are one of the causes of acute and chronic kidney damage. The aim of the study was to compare some serum biochemical parameters of rats at two and six weeks of therapeutic doses of nalbuphine and to compare them after early withdrawal. This study was performed on 45 white sexually mature male rats weighing 180–260 g. In the serum of experimental animals’ total protein, urea, creatinine, calcium and phosphorus were determined. Statistical calculations were performed using RStudio v. 1.1.442 and R Commander v.2.4-4. Serum total protein in the study groups had small fluctuations in control, but there was a tendency to decrease after 2 weeks of nalbuphine administration and to increase both from the second to the sixth week and with the withdrawal of nalbuphine in the early stages of the experiment. Serum creatinine increased from the second to the sixth week of the experiment both with six weeks of administration (2.23 times higher than control values) and with two weeks of nalbuphine with the subsequent four-week withdrawal (1.63 times higher than control values). Serum urea increased after 2 weeks and 6 weeks of nalbuphine administration, with mean values 6.22 ± 0.47 mmol/l and 7.63 ± 0.74 mmol/l respectively. However, after drug withdrawal the indicator decreased to the control group. Serum calcium in rats decreased after 2 weeks of the experiment to the value (2.16 ± 0.09 mmol/l). After 6 weeks of nalbuphine administration, as after drug withdrawal, serum calcium increased to the level of the control group and was 2.39 ± 0.09 mmol/l and 2.46 ± 0.05 mmol/l respectively. The serum phosphorus index increased after 2 weeks of the experiment and was 2.42 ± 0.05 mmol/l. After 6 weeks of nalbuphine administration and after drug withdrawal, the value decreased to 2.12 ± 0.12 mmol/l and 2.00 ± 0.05 mmol/l respectively. Conclusions. The use of therapeutic doses of nalbuphine for two and six weeks, as well as after drug withdrawal does not affect the concentration of serum total protein of rats. As a result of two- and six-week use of the drug, an impaired renal nitrogen excretion is observed, and hence the structure of the renal corpuscles is altered, which also takes place during four weeks of drug withdrawal after two weeks of use, as evidenced by an increase in urea and creatinine after two weeks of nalbuphine and significant increase in creatinine after drug withdrawal. Two-week administration of nalbuphine leads to hypocalcemia and hyperphosphatemia, indicating impaired renal reabsorption function, while a subsequent four-week administration and withdrawal leads to normalization of these parameters, except for phosphorus, which is reduced.

Экспрессия miR-181a и miR-146a при преждевременной овариальной недостаточности аутоиммунной этиологии

Objective. To study the expression of miR-181a and miR-146a in the blood serum obtained from an experimental animal model of autoimmune oophoritis on different days of the experiment to assess their role in primary ovarian insufficiency (POI) of autoimmune etiology. Materials and methods. Experimental animals were mature female outbred rats. The study group (n = 25) included the experimental animal model of autoimmune oophoritis, the control group (n = 20) – intact animals. Morphometry of ovarian samples with estimating the average number of follicles, corpora lutea and atretic corpora lutea was performed. Enzyme immunoassay was used to study the concentration levels of serum anti-Mullerian Hormone (AMH) and anti-ovarian antibodies (AOA). MicroRNAs were isolated from serum according to a standard protocol; a reverse transcription and real-time polymerase chain reaction were performed using specific primers, and the relative expression of miR-181a and miR-146a was calculated by the Pfaffle method. Results. The expression of miR-146a on different days of the experiment remained statistically insignificant, including when compared to the control group. The expression of miR-181a in the blood serum of experimental animals on day 15 was statistically significantly higher than similar values on days 5, 10, 30 and 60 and in the control group. Conclusion. MiR-181a showed high diagnostic value in POI of autoimmune etiology in the experiment performed and can serve as an early marker in the diagnosis of ovarian insufficiency. The role of miR-146a as a marker of POI of autoimmune etiology has not been experimentally confirmed. Key words: autoimmune oophoritis, microRNA, experiment, premature ovarian insufficiency

The dynamics of cytolytic syndrome parameters in the period of late manifestations of cranioskeletal trauma in case of concomitant chronic hepatitits and the effectiveness of armadine correction

Relevance. Cranioskeletal trauma has the most severe lesions and is accompanied by significant mortality. The impact of comorbidities caused by the cardiovascular system, in particular diffuse lesions of internal organs are not fully investigated under this condition. A common mechanism of pathogenesis of these pathological processes is the development of cytolytic syndrome.
 Goal of study: is to determine the dynamics of markers of cytolysis in the period of late manifestations of cranioskeletal trauma in conditions of concomitant chronic hepatitis and to evaluate the effectiveness of armadine correction.
 Materials and methods. Cranioskeletal trauma, chronic hepatitis, and its combinations were modelled in separate groups of nonlinear white male rats. Armadine at a dose of 20 mg/kg intraperitoneally was used to correct the group with combined lesions. The activity of alanine and aspartate aminotransferases was determined in the serum of experimental animals on 14, 21, 28 and 35 days after injury.
 Results and its discussion. The application of cranioskeletal trauma in the late manifestations of traumatic disease is accompanied by increased cytolysis. The activity of alanine and ampartate aminotransferase in the serum of injured animals is increased after 14 days of the post-traumatic period and continues to increase up to 21 days with a subsequent decrease. Moreover, up to 35 days only the activity of aspartate aminotransferase reaches the level of control, while the activity of alanine aminotransferase continues to remain significantly higher. The use of armadin under these conditions from the 21st day of the experiment causes a decrease in the changes of the studied parameters compared with the group of animals without correction.
 Conclusions. The application of cranioskeletal trauma on the background of chronic hepatitis in the late manifestations of traumatic disease is accompanied by a greater change of cytolysis than in the group of injured animals without concomitant liver damage. The use of armadine causes a statistically significant decrease in the intensity of the detected disorders, starting from 21 days of the experiment.

Use of probiotics for formation of microflora of gastrointestinal tract of calves

The results of the use of probiotic strains of microorganisms of the Bacillus family for the correction and formation of the microflora of the gastrointestinal tract and the impact on metabolism in calves are presented.
 The aim of the study. To analyze the effect of probiotics on the microflora of the gastrointestinal tract in calves and biochemical parameters of blood in calves up to one month.
 Materials and methods. The research was conducted during 2020 in the conditions of Ukrainian farms for cattle breeding. Five experimental groups of five one-week-old calves were formed in each and one control group. Calves were kept separately in the same conditions on the same diet, but with feeding together with colostrum substitute probiotics of five grams per animal: Bacillus amyloliquefaciense, Bacillus mucilaginosus, Bacillus coagulans, Bacillus megaterium, Bacillus pumilus. The strains are deposited and produced by “Kronos Agro” Ukraine.
 Results. It was found that as a result of studies when feeding calves B. coagulans, B. pumilus and B. mucilaginosus the number of Lactobacillus sp. was 80 % higher than in the control group. The level of opportunistic pathogens in the experimental group with B. coagulans had minimal values. Animals in the group where B. mucilaginosus was given had a higher amount of Candida - up to 300 CFU/g and Enterobacteriaceae – 200 CFU/g; which is 50 % less compared to control groups, but more than in the experiment with B. coagulans. According to the results of biochemical examination of blood serum in calves, the absence of toxic effects of probiotic strains: Bacillus amyloliquefaciense, Bacillus mucilaginosus, Bacillus coagulans, Bacillus megaterium, Bacillus pumilus on the internal organs of animals was established.
 Conclusions. It was found that the maximum positive effect on the microflora of the gastrointestinal tract of calves up to 30 days of age had B. coagulans (1×109) when fed at a dose of 5 g per animal. The amount of Lactobacillus sp. was the maximum and reached 800 CFU/g, which is 80 % more than in the control group. At the same time, the level of opportunistic pathogens in the experimental group with B. coagulans had minimal indicators and was: Clostridium by 20 %, Escherichia coli – by 70 %, Enterobacteriaceae, Staphylococcus and Candida – 100 % less than the control.
 In the study of biochemical parameters, it was found that the activity of enzymes, protein and glucose levels in the serum of experimental animals fed with B. coagulans were within the physiological norm, indicating a normal metabolic process and no toxic effects.

Combination therapy using nitro compounds improves the efficacy of experimental Chagas disease treatment.

Drug combinations have been evaluated for Chagas disease in an attempt to improve efficacy and safety. In this line, the objective of this work is to assess the effects of treatment with nitro drugs combinations using benznidazole (BZ) or nifurtimox (NFX) plus the sulfone metabolite of fexinidazole (fex-SFN) in vitro and in vivo on Trypanosoma cruzi infection. The in vitro interaction of fex-SFN and BZ or NFX against infected H9c2 cells by the Y strain was classified as an additive (0.5⩾ΣFIC<4), suggesting the possibility of a dose reduction in the in vivo T. cruzi infection. Next, the effect of combining suboptimal doses was assessed in an acute model of murine T. cruzi infection. Drug combinations led to a faster suppression of parasitemia than monotherapies. Also, the associations led to higher cure levels than those in the reference treatment BZ 100 mg day−1 (57.1%) (i.e. 83.3% with BZ/fex-SFN and 75% with NFX/fex-SFN). Importantly, toxic effects resulting from the associations were not observed, according to weight gain and hepatic enzyme levels in the serum of experimental animals. Taken together, this study is a starting point to explore the potential effects of nitro drugs combinations in preclinical models of kinetoplastid-related infections.

STUDY OF THE ACTIVITY OF SOME RAT SERUM ENZYMES IN THE DYNAMICS OF EXTRAHEPATIC CHOLESTASIS

Extrahepatic cholestasis occurs with mechanical obstruction of the main extrahepatic or main intrahepatic ducts. The most common cause of extrahepatic cholestasis is common bile duct stones. This article describes hypertension in the bile ducts and acholysis developing in cholestasis, which cause serious functional and morphological changes in the liver and lead to the rapid growth of liver insufficiency. Thus, in the dynamics of extrahepatic cholestasis development there are significant shifts in blood serum of experimental animals, manifested by hyperfermentemia. These changes indicate the involvement of the liver in the pathological process, which naturally requires their correction.

INFLAMMATION AND IMPACT OF VINCRISTINE AND ENTEROSORPTION USE IN CHEMICALLY INDUCED COLON CARCER IN RATS

Background. The increasing incidence of colon malignant tumors is one of the most urgent matters of contemporary medicine. In the study of carcinogenesis of the colon the attention is paid to the state of the body’s immune system and activation of inflammatory processes in experimental animals.&#x0D; Objective. The aim of the study was to estimate the level of markers of inflammation in the serum of experimental animals with chemically induced carcinogenesis and their dynamics in case of administration of the cytostatic Vincristine secondary to AUT-M carbon enterosorbent.&#x0D; Methods. The study was performed on white male rats. Animals were modeled for colon cancer by administration of 1.2-dimethylhydrazine hydrochloride at a dose of 7.2 mg/kg body weight for 30 weeks. AUT-M enterosorbent was administered intragastrically daily during 7 and 21 days after modeling of carcinogenesis at a dose of 1 ml of suspension (corresponding to 0.2 g of drug weight) per 100 g of animal body weight. The antitumor drug was administered to the animals with induced carcinogenesis intragastrically daily during 14 days at a dose of 0.23 mg/kg of body weight after a 21-day detoxification therapy. The activity of inflammatory processes was evaluated by the content of pro-inflammatory interleukin 6 and anti-inflammatory interleukin 4, C-reactive protein in the serum of experimental animals.&#x0D; Results. It was established that introduction of 1.2-dimethylhydrazine hydrochloride in the rats caused changes in the cytokine profile and the content of C-reactive protein. In the affected animals an increase in the content of pro-inflammatory interleukin 6, C-reactive protein, as well as a decrease in the content of anti-inflammatory interleukin 4 was evidenced in all periods of the study. AUT-M enterosorbent contributed to normalization of these parameters. The cytostatic Vincristine had a negligible effect on development of inflammatory processes in the studied animals.&#x0D; Conclusions. In cases of induced carcinogenesis, an imbalance in the content of pro- and anti-inflammatory cytokines, an increase in the content of acute-phase C-reactive protein was established. The positive effect of the cytostatic Vincristine secondary to a previous detoxification therapy with AUТ-M sorbent during a progressive development of inflammatory processes in the presence of modeled carcinogenesis was evidenced.

Активність трансаміназ і вміст білірубіну у крові щурів за введення антибіотика енрофлоксацину, нанополімеру ПЕГ-400 та їх комплексу

The large-scale production of antibiotics able to the targeted drug delivery to the affected tissues and target cells is relevant for ensuring an increase in the effectiveness of the humans and animals treatment. The aim of this study is to evaluate the effect of the antibiotic enrofloxacin applied alone or in combination with the nanopolymer PEG-400 on the activity of the transamination enzymes (ALT, AST) and the concentration of bilirubin in the rats’ serum. The complex enrofloxacin-PEG-400 was obtained by the reaction of enrofloxacin chloride with PEG-400. Four groups of 12 rats each were studied; there were three experimental groups and a control one. Control group rats were injected intramuscularly with saline, the first experimental group with the antibiotic enrofloxacin, the second with the nanopolymer PEG-400, the third with the complex enrofloxacin-PEG-400. The volume of administrated drugcorresponded to the dose of enrofloxacin for animals’ treatmentand it was 0.03 ml per 200 g of rat weight for all groups. The drugs were being administered for four days, every day. Decapitation of animals was carried out in 7, 14 and 12 days after drug administration. In the blood of animals the AST / ALT activity and their ratio (de Ritis Ratio) have been studied, as well as the total bilirubin. Seven days afterthe drugs introduction,the ALT activity in the serum of the twofirst experimental groups of rats, injected with the antibiotic enrofloxacin in its traditional form and nanopolymer PEG-400, corresponded to control values, whereas theALT activity in the blood of animals afterthe administration of nanopolymer PEG-400 and complexwas lower than control values.At the same time, the AST activity in the serum of the first experimental group was at the control level.As for the others ones,nanopolymer PEG-400 and complexwere administered,the higher rates could be due to active penetration of these substances into cells and mitochondria, where this enzymewas localized. De Ritis Ratioincreased, so it certifies the active elimination of AST in the blood. The bilirubin level in the serum of experimental animals was higher than the level of control. Such changes in the blood parameters of experimental ratscan be considered asa minor disturbance of the liver structure and function. On day 14 of the experiment, in comparison with day 7, the ALT activity in the serum of all rats groups resulted in a significant reduction, except for the first experimental group. The lowest activity rates were in the third experimental group and they corresponded to physiological values.During the experiment, the AST activity in the blood of all animals increased in comparison with the first study, but it didn’t go beyond physiological fluctuations.The maximum activity was determined after the administration of the traditional form of enrofloxacin. As for other experimental groups, they rates were a little lower.However, it can be assumed that enrofloxacin in its traditional formcan enter the bloodstream from the place of administration much more slowly than nanopolymerPEG-400 and complex enrofloxacin – nanopolymerPEG-400 and it can be metabolized in the livertissue.This fact can be described as an improvementof the speed of penetration and accumulation in cells of the antibiotic enrofloxacin combined with the nanopolymer PEG-400 for its transportation. De Ritis Ratio was higher in the studied rats. At the same time, the bilirubin level of all experimental animals decreased, but remained higher, as compared to controls. 21 days after the administration of the test substances, the ALT activity in the animals’ blood was going down. In the third experimental group, as well as in the control, the enzyme activity level in the blood was within physiological fluctuations, while in rats, injected with the antibiotic enrofloxacin and PEG-400 separately, the ALT activity was higher. On day 21of the experiment, the AST activity in the blood of all animals was at the physiological level. Meanwhile, the maximum value of the enzyme after the use of PEG-400 was a little lower than after the administration of the antibiotic enrofloxacin alone.At the same time it was lower after the complex enrofloxacin with nanopolymer PEG-400 administration.According to the dynamics of ALT/AST activity in the blood of animals, it may be concluded that the recovery of liver cells structure is faster in rats, injected with the complex enrofloxacin with nanopolymer PEG-400. The total bilirubin in the blood of all experimental groups has decreased. Thus, the obtained blood parameters can certify that three weeks after the administration of the antibiotic enrofloxacin, nanopolymer PEG-400 and complex enrofloxacin with PEG-400 the structure and the functional state of liver cellsrenewal process is going to start. The best results have been obtained after the use of complex enrofloxacin with PEG-400. In the further research it is planned to study the effect of the enrofloxacin – nanopolymer PEG-400 complex on the functional state and the structure of other organs and systems of the body.

CORRECTION OF STRESS-INDUCED HORMONAL CHANGES BY THE RIL-2 MEDICATION IN EXPERIMENTAL TRAUMATIC BRAIN INJURY

Traumatic brain injury (TBI) leads to steady stress reaction that may substantially influence on posttraumatic period. Stress changes concentration of sex hormones too, that play role of neurosteroids in CNS. The aim of the study was to detect changes in corticosterone and testosterone concentration in serum of experimental animals during recovery period after TBI. And also to find possibility of disturbance correction by cytokine preparation rIL-2 (ronkoleukin). After TBI both experimental and control group of animals get daily injections of recombinant human interleukin-2. TBI decreased the level of corticosterone on 30%, the injection of rIL-2 increased it. Testosterone slightly changed after TBI. But it’s level increased when after TBI the injections of rIL-2 were done. The obtained results can be considered as evidence of the stress reaction normalization in animals after TBI and under the conditions of their treatment with rIL-2.

Hydrocarbon metabolism in mice during preclinical studies of modified starch

Preclinicl studies of new nutritional supplements include the establishment of not only harmlessness, lethal or semi-lethal dose, but also biochemical signs of the laboratory animals at certain doses of the substance. New food additives include modified starch. Of scientific interest are the studies of white mice hydrocarbon metabolism considering the safety of modified starch. The study of modified starch safety was performed according to the established requirements. Eighteen 2-month-old laboratory mice (females) were used for the study, with an average body weight of 19–21 g. Groups were formed using the randomization method. Each group consisted of six experimental animals. Experiments on the harmlessness of modified starch on white mice were performed according to the scheme, where animals from the control group were intramuscularly administered 0.3 cm3 of 0.9% NaCI solution. Mice from the first experimental group were led with a 5.0% solution of modified starch. Animals from experimental II were guided by a 10.0% solution of modified starch. The volume of solutions of modified starch was 0.3 cm3. Blood serum of laboratory mice was tested for glucose, lactic and pyruvic acid levels, and the liver was used to determine glycogen level. All those actions were performed according to current bioethics requirements. It was experimentally proven that on the eleventh day from the beginning of injecting the 0.3 cm3 5.0% and 10.0% solution of modified starch in the blood serum of the laboratory mice glucose content did not differ from those obtained in control animals, which were injected with saline. When using the test solutions, itt was found that the levels of lactic and pyruvic acid in the serum of experimental animals were within the physiological standards. In those mice that were injected intragastrically the increased doses of modified starch the level of glycogen in the liver was about 39.7–40.1 g/kg which corresponded to physiological standards. Thus, a single injection of 0.3 cm3 5.0% and 10.0% solution of modified starch does not cause disorder of hydrocarbon metabolism in mice.

Influence of B. bovis hairworms on biochemical parameters of blood of patients with bovicolosis of calves

The purpose of the work was to identify the features of the pathological process and changes in the main indicators of blood of cattle for the damage by its hairworms B. bovis. Scientific researches were conducted in the conditions of the Ploskivske BPPZ of the Brovarsky District of the Kyiv Region and the Laboratory of Biochemical Research of the National Cancer Institute. To do this, two groups of young cattle, black-and-white breeds up to one year old with 7 heads in each, were created. The experimental group included calves spontaneously invasive with hairworms. The control group was free of animal parasites. For this, they were previously, in two months, subjected to two-fold treatment with Insecticide Tactics (MSD, the Netherlands). Selected animals on the principle of analogues. All calves were in the same sanitary and hygienic conditions and received the same feed according to the diet. It was found that the glucose content in a sick young bovine animal was 6.25% (P &lt; 0.5) higher than that in calf-free calves. There was a probable difference in the level of urea in the blood of animals, where the average level in the infected calves was 12.69% (P &lt; 0.001) higher than in the control animals. The tendency to decrease by 15.31% (P &lt; 0.5) was found in the average values of serum creatinine in calves in sick animals compared with the group of animals that served as controls and were free from parasites. In blood serum, calves infected with hairworms was found a slightly higher (4.25%, P &lt; 0.5) level of total bilirubin than those found free from parasites. Indicators of serum cholesterol levels in blood serum of experimental animals were within normal limits. However, it was significantly lower by 14.71% (P &lt; 0.05) in animals affected by parasites. The albumin-globulin ratio in our studies in invasive calves was 9.8% (P &lt; 0.01) higher than that of control animals. In experimental calves with clinical manifestations of bovycolous, the average index of alkaline phosphatase content was 17.58% (P &lt; 0.5) lower than its mean value in blood serum free of invasion by parasites of animals. Thus, among the biochemical parameters of serum of blood of invasive calves, a probable increase in urea and albumin to globulin ratios and a decrease in cholesterol was observed, indicating a negative effect of the hairworms on the organism of animals.

Morphological and biochemical indexes of blood of domestic sheep for 3 periods of lambing in the conditions of the Central Yakutia

The results of morphological and biochemical parameters of blood of ewes in the period of pregnancy and after childbirth for 3 periods of lambing in Yakutia are presented. It was found that an increase in the content of red blood cells in the blood after childbirth caused an increase in the concentration of hemoglobin in sheep of both breed groups. At the same time statistically significant increase of hemoglobin concentration was revealed in 1 and 3 lambs. Thus, the increase in the value of the studied index in 1 lamb, in sheep of the Romanov breed was 117.5 ± 1.12 g/l or 3.8 % ( p≥ 0.95), in TRANS - Baikal fine - wool - 118.8 ± 0.99 g/l (P ≥ 0.99), in 3 lambs: in Romanov sheep-119.4 ± 0.91 g/l or 2.8 % ( P≥ 0.95 ), in TRANS-Baikal-119.9 ± 1.99 g/l or 8.6 % ( P≥ 0.95).0.95). According to the biochemical parameters of blood it was found that during the period of pregnancy the ewes of both breed groups differed by the maximum level of total protein for the periods of research. After delivery, there was a decrease in the concentration of total protein in the serum of experimental animals. Thus, the level of total protein reduction in 1 lamb - in sheep of the Roma breed was 7.55 g/l or 10.9% (p ≥ 0.99), in TRANS-Baikal – 0.8 g/l or 1.1 %; in 2 lambs – 6.39 g/l (9.09%) with a significant difference (p ≥ 0.99), and 0.58 g/l (0.78 %); in 3 lambs – 3.13 g/l (4.49%) (P ≥ 0.99), and 1.14 g/l (p 1.45 %). At the same time, attention is drawn to the fact that the level of total blood protein quite clearly expresses the physiological state of the Romanov sheep breed and is less susceptible to changes in the TRANS-Baikal sheep. In General, biochemical parameters of ewes for 3 periods of lambing were within physiological norms and significant deviations were not revealed and are caused by the manifestation of the General biological regularity of the physiological state of sheep during pregnancy and after childbirth.

Cardioprotective Effect of Rhapontigenin in Isoproterenol-Induced Myocardial Infarction in a Rat Model

Background/Aims: Rhapontigenin (RPG) is a stilben derivative and is known to bear several effects such as antiallergic, anticoagulative, hypoglycemic, antiangiogenic, and purgative. The investigation was conducted to evaluate the cardioprotective efficacy of RPG in rats having acute myocardial infarction (MI) induced by isoproterenol (ISO). Methods: Animals were divided into 6 groups: group I (control group), group II (ISO-treated), group III (1.0 mg/kg/day RPG and ISO-treated), group IV (2.5 mg/kg/day RPG and ISO-treated), group V (5.0 mg/kg/day RPG and ISO-treated), and group VI (treated with RPG 5.0 mg/kg/day). Various cardiac stress markers, including infarct size and heart/body weight index, were investigated in animals with ISO-induced MI, such as inducible nitric oxide synthase (iNOS), creatinine kinase (CK), lactate dehydrogenase (LD), cardiac troponin-T (CTT), superoxide dismutase (SOD), and malondialdehyde. INOS, p38, caspase-3, and connexin 43 expressions were analyzed in animals. Inflammatory mediators, tissue necrosis factor-α (TNF-α) and interleukin-6 (IL-6), were detected in serum of experimental animals. Results: Group I animals indicated normal levels of biochemical parameters, whereas group II animals indicated high levels of these parameters and successful induction of MI. Pretreatment of animal groups III, IV, and V with RPG revealed amelioration of infarct size, heart/body weight index, CK, LD, CTT in rats, whereas group VI animals were treated only with RPG (5.0 mg/kg/day) and not with ISO. Levels of TNF-α, IL-6, MD, SOD, p38, and iNOS expressions were significantly downregulated by RPG administration (5.0 mg/kg/day). Conclusion: RPG ameliorates MI caused by ISO in rats and provides cardioprotective effect, via anti-inflammatory, antioxidant, and antiapoptotic effect.

Multiple organ toxicity of Datura stramonium seed extracts.

Datura stramonium seed ranks top among major plants commonly abused as drug in Nigeria. The present study therefore sought to unravel the target organs of toxicity as well as underscore the role of extraction solvent in the toxicity of Datura stramonium seed. Twenty male Wistar rats were randomly placed into four groups (I-IV) of five animals per group. Group I served as the control and was administered with distilled water only, while groups II, III and IV animals received 50 mg/kg body weight of aqueous, methanolic and diethylether extracts of Datura stramonium seeds by oral gavage for 14 days. Specific biomarkers of toxicity such as aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and lipid peroxidation (MDA) were estimated in the liver, brain, kidney and heart homogenates as well as serum of experimental animals. Lipid profile and activity of antioxidant enzymes such as superoxide (SOD) and catalase were determined in selected tissues while creatinine and urea were estimated in the kidney and serum. Results indicated that Datura stramonium seed extract generally caused a significant decrease in ALT, AST, ALP and MDA in selected tissue homogenates while these parameters increased significantly in the serum relative to the control group. Lipid profile was significantly deranged across selected tissues while SOD and catalase activity were significantly decreased relative to control. Put together, toxicity of Datura stramonium seed extract is diverse depending on the organ involved and solvent used for its extraction. Therefore, illicit use of Datura stramonium seeds as drugs among young people should be discouraged.

Особливості обміну речовин чистопородного і помісного молодняку свиней

Особливості обміну речовин чистопородного і помісного молодняку свиней

Abstract A09: Development of epigenetic-based strategies for blocking breast cancer growth, invasion, and metastasis in vitro and in vivo

Abstract DNA hypomethylation has been implicated in the coordinated targeting of various signaling pathways involved in tumor growth and metastasis. In the current study, through various in vitro and in vivo assays, we have examined the plausibility of using universal methyl donor S-adenosyl methionine (SAM) to block breast cancer development, growth, and metastasis in our xenograft and transgenic models of breast cancer. Treatment of highly invasive human triple-negative breast cancer (TNBC) cell lines MDA-MB-231 and Hs578T with SAM resulted in a significant dose-dependent decrease in cell proliferation, invasion, migration, colony formation, and increased apoptosis in vitro. Affymetrix gene expression array and real-time PCR (qPCR) validated the ability of SAM to decrease the expression of several genes implicated in cancer progression in MDA-MB-231 cells. For the in vivo studies, MDA-MB-231 cells expressing green fluorescent protein (MDA-MB-231-GFP) were inoculated into female CD-1 nude mice via mammary fat pad. From day three post tumor cell inoculation, animals were treated with SAM (0.8-1.6 mg/day) or vehicle alone as control via daily oral gavage, and tumor volume was determined at weekly intervals for 10 weeks. SAM treatment caused a significant dose-dependent decrease in tumor volume and GFP-positive metastasis to lungs, liver, and spleen in experimental animals compared to vehicle-treated controls. Analysis of RNA from primary tumors by qPCR showed the ability of SAM to cause a marked decrease in the expression of several prometastatic and EMT pathway genes. Pyrosequencing of tumoral DNA from control and experimental animals showed that SAM treatment causes a significant increase in the percentage of CpG methylation at the promoter region of several cancer-related genes which were seen to be downregulated in the qPCR assay. We next determined the effect of SAM in MMTV-PyMT transgenic mouse model of breast cancer where SAM treatment resulted in a significant delay in the development of mammary tumors and decreased tumor growth in experimental animals as compared to vehicle-treated controls. SAM was found to bioavailable in the serum of experimental animals as determined by mass spectrometry and no notable adverse side effects were seen, including any change in animal behavior. Results from these studies provide compelling evidence for the therapeutic potential of SAM in breast cancer to provide the rationale for initiating clinical trials with SAM in patients with breast and other common cancers as monotherapy or in combination setting with current therapeutic agents to reduce cancer-associated morbidity and mortality. Note: This abstract was not presented at the conference. Citation Format: Niaz Mahmood, David Cheishvili, Ani Arakelian, William J. Muller, Imrana Tanveer, Haseeb Khan, Moshe Szyf, Shafaat A. Rabbani. Development of epigenetic-based strategies for blocking breast cancer growth, invasion, and metastasis in vitro and in vivo [abstract]. In: Proceedings of the AACR International Conference held in cooperation with the Latin American Cooperative Oncology Group (LACOG) on Translational Cancer Medicine; May 4-6, 2017; São Paulo, Brazil. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(1_Suppl):Abstract nr A09.

Effects of Vitamin D3 on the NADPH Oxidase and Matrix Metalloproteinase 9 in an Animal Model of Global Cerebral Ischemia.

Decreased blood flow in the brain leads to a rapid increase in reactive oxygen species (ROS). NADPH oxidase (NOX) is an enzyme family that has the physiological function to produce ROS. NOX2 and NOX4 overexpression is associated with aggravated ischemic injury, while NOX2/4-deficient mice had reduced stroke size. Dysregulation of matrix metalloproteinases (MMPs) contributes to tissue damage. The active form of vitamin D3 expresses neuroprotective, immunomodulatory, and anti-inflammatory effects in the CNS. The present study examines the effects of the vitamin D3 pretreatment on the oxidative stress parameters and the expression of NOX subunits, MMP9, microglial marker Iba1, and vitamin D receptor (VDR), in the cortex and hippocampus of Mongolian gerbils subjected to ten minutes of global cerebral ischemia, followed by 24 hours of reperfusion. The ischemia/reperfusion procedure has induced oxidative stress, changes in the expression of NOX2 subunits and MMP9 in the brain, and increased MMP9 activity in the serum of experimental animals. Pretreatment with vitamin D3 was especially effective on NOX2 subunits, MMP9, and the level of malondialdehyde and superoxide anion. These results outline the significance of the NOX and MMP9 investigation in brain ischemia and the importance of adequate vitamin D supplementation in ameliorating the injury caused by I/R.

Methyl donor S-adenosylmethionine (SAM) supplementation attenuates breast cancer growth, invasion, and metastasis in vivo; therapeutic and chemopreventive applications.

DNA hypomethylation coordinately targets various signaling pathways involved in tumor growth and metastasis. At present, there are no approved therapeutic modalities that target hypomethylation. In this regard, we examined the therapeutic plausibility of using universal methyl group donor S-adenosylmethionine (SAM) to block breast cancer development, growth, and metastasis through a series of studies in vitro using two different human breast cancer cell lines (MDA-MB-231 and Hs578T) and in vivo using an MDA-MB-231 xenograft model of breast cancer. We found that SAM treatment caused a significant dose-dependent decrease in cell proliferation, invasion, migration, anchorage-independent growth and increased apoptosis in vitro. These results were recapitulated in vivo where oral administration of SAM reduced tumor volume and metastasis in green fluorescent protein (GFP)-tagged MDA-MB-231 xenograft model. Gene expression analyses validated the ability of SAM to decrease the expression of several key genes implicated in cancer progression and metastasis in both cell lines and breast tumor xenografts. SAM was found to be bioavailable in the serum of experimental animals as determined by enzyme-linked immunosorbent assay and no notable adverse side effects were seen including any change in animal behavior. The results of this study provide compelling evidence to evaluate the therapeutic potential of methylating agents like SAM in patients with breast cancer to reduce cancer-associated morbidity and mortality.