Animals In Control Group Research Articles

Changes in the Cyto- and Fibroarchitectonics of the Cerebellar Cortex in Rats Subjected to Extreme Physical Activity.

Physical overexertion surpassing the functional capacity of the nervous system causes the hyperactivation of the neural structures of the cerebellum. In turn, it causes the depletion of intracellular resources and progressive structural changes in cerebellar cells and fibers. These degenerative changes may lead to cerebellar dysfunction, including the worsening of coordination, balance, and motor functions. In order to maintain the health and functioning of the cerebellum and the nervous system in general, one needs to avoid physical overexertion and have enough time to recover. Three major types of Purkinje cells were identified in control group animals. After the forced swimming test, animals had significant morphological changes in pyriform cells, granule cells, internuncial neurons, and neuroglial cells. In particular, the extreme degeneration of granule cells was manifested via their fusion into conglomerates. These changes demonstrate that neurodegeneration in the cerebellum takes place in response to physical overexertion.

Preclinical evaluation of 225Ac-labeled minigastrin analog DOTA-CCK-66 for Targeted Alpha Therapy.

Treatment studies were performed (3 groups, n = 5, AR42J tumor-bearing 394-NOD SCID mice). Control group animals were injected with [68Ga]Ga-DOTA-CCK-66 (1.1MBq, PET/CT imaging), while treatment group animals received a single dose of either [177Lu]Lu-DOTA-CCK-66 (37MBq, radioligand therapy (RLT)) or [225Ac]Ac-DOTA-CCK-66 (37kBq, TAT). All animals' tumor volume and body weight were monitored twice a week until end-point criteria were reached. Blood samples were evaluated (VetScan VS2, Abaxis) once mice were sacrificed. Upon treatment, an initial decline in tumor volume, followed by a significantly delayed tumor growth of treated cohorts, was observed. Mean survival of 177Lu- as well as 225Ac-treated animals was increased by 3- (37 ± 3 d) and 4.5-fold (54 ± 6d), respectively, when compared to non-treated animals (12 ± 3d). Blood sample analysis did not indicate toxic side effects to the liver, kidney, or stomach upon 177Lu and 225Ac-treatment. We demonstrated a substantial therapeutic efficacy of 177Lu- and 225Ac-labeled DOTA-CCK-66. As expected, treatment with the latter resulted in the highest mean survival rates. These results indicate a high therapeutic potential of 225Ac-labeled DOTA-CCK-66 for TAT in MTC patient management.

Gastroduodenal effects of leflunomide under experimental adjuvant-induced rheumatoid arthritis and comorbid arterial hypertension

Use of leflunomide (LEF), a synthetic immunosuppressor, for the treatment of rheumatoid arthritis (RA), has become an innovation in the management of the disease. Despite the widely and successfully use of the LEF for RA treatment, its pharmacological properties and side effects are not well understood. The aim of the study was to examine the gastroduodenal effects of prolonged enteral use of the LEF in adjuvant-induced RA and comorbid arterial hypertension (AH). The experiments were carried out on male and female mature nonlinear rats, which were divided into five groups. The animals of control group (n = 10) were treated with starch suspension, the animals of the second group (n = 15) were injected with complete Freund’s adjuvant – adjuvant-induced arthritis (AA), the third group (n = 15) AA and comorbid AH (salt-loading model), the fourth group (n = 15) consisted of animals with AA treated with LEF, the fifth group (n = 15) – animals with comorbid condition (AA + AH), which were treated by LEF. Administering of starch-LEF suspension into the stomach was carried out through a special metal probe to rats with adjuvant-induced arthritis and comorbid pathology once daily for the first 3 days at a dose of 15 mg/kg (shock dose) and then at a dose of 1.5 mg/kg (therapeutic dose) for the next 53 days. After finishing the experiments, gastroduodenal toxic effects of the drug were determined according to the frequency of rats with pathological lesions (hyperemia, petechiae, edema, ulcers) and intensity of the damages using a point scale. It has been established that prolonged use of the LEF leads to high levels of gastroduodenal toxicity in rats with adjuvant-induced arthritis and combined pathological condition: an increase in the frequency of rats with gastroduodenal mucosal pathology and intensity of various types of damages (petechiae, edema, ulcers). LEF induced higher prevalence of duodenal than gastric toxicity in rats with both adjuvant-induced arthritis and arterial hypertension comorbidity. Thus, gastroduodenal mucosa pathology after prolonged use of LEF in adjuvant-induced arthritis and comorbid state increases a need for adjuvant therapy to prevent such adverse effects. Moreover, further pharmacological research in order to introduce appropriate drugs is recommended. Furthermore, conducting preclinical and clinical studies to understand the pharmacokinetics, mechanisms of action, and interaction of LEF, in particular, with antihypertensive and gastroprotective agents seem necessary. Implementation of the results of the study in healthcare practice will help to optimize the treatment regimen for rheumatoid arthritis and comorbidities.

Lining the small intestine with mycobacteriophages protects from Mycobacterium avium subsp. paratuberculosis and eliminates fecal shedding

Johne's disease (JD), a chronic, infectious enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP), affects wild and domestic ruminants. There is no cure or effective prevention, and current vaccines have substantial limitations, leaving this disease widespread in all substantial dairy industries causing economic, and animal welfare implications. Mycobacteriophages (MPs) have been gaining interest in recent years and are proposed as a promising solution to curtailing MAP infection. Using a well-validated infection model, we have demonstrated the preventative potential of MPs to protect dairy calves against MAP infection. Calves were supplemented daily with a phage cocktail from birth till weaning at 2 m of age and inoculated with MAP at 2 wk of age. Infection status was measured for 4.5 mo through blood, fecal, and postmortem tissue samples. Our findings highlight the remarkable efficacy of orally administered MPs. Notably, fecal shedding of MAP was entirely eliminated within 10 wk, in contrast to the infected control group where shedding continued for the entirety of the trial period. Postmortem tissue culture analysis further supported the effectiveness of MPs, with only 1 out of 6 animals in the phage-treated group testing positive for MAP colonized tissues compared to 6 out of 6 animals in the infected control group. Additionally, plaque assay results demonstrated the ability of phages to persist within the intestinal tract. Collectively, these results underscore the potential of orally administered MP cocktails as a highly effective intervention strategy to combat JD in dairy calves and by extension in the dairy industry.

Electroacupuncture at Zusanli regulates the pathological phenotype of inflammatory bowel disease by modulating the NLRP3 inflammasome pathway.

This study sought to explore the effect of electroacupuncture (EA) intervention at Zusanli (ST36) acupoint on modulating the NLRP3 inflammasome pathway for treating inflammatory bowel disease (IBD). C57BL/6 mice were administrated with 3% dextran sulfate sodium (DSS) to construct the IBD model. DSS mice were then administrated with EA (10 Hz, 1.5 mA) at ST36 for 7 days or intragastric administration of sulfasalazine (SASP) each day during the entire course. The control group animals were administered with distilled water. Then, partial least squares discriminant analysis revealed differences in the relative content of metabolites. The pathological changes of colon and spleen tissues were observed by H&E and immunohistochemistry (IHC) staining. qPCR determined the mRNA expression levels, while ELISA and western blot analysis determined the protein expression. Compared with the control groups, DSS-induced decreases of body weight were reversed after EA stimulation at ST36 or SASP treatment. The DAI of DSS mice was significantly higher relative to the control groups, whereas the DAI of DSS mice were decreased after EA stimulation at ST36 or SASP treatment. The intestinal weight/length ratio increased significantly in DSS groups; however, EA at ST36 significantly improved the macroscopic/microscopic characteristics and the weight and length of the colon. EA reversed inflammation and leukocyte infiltration and normalized the elevated levels of IL-1β, IL-18, and NLRP3. Furthermore, EA improved the expression levels of ZO-1, occludin, and claudin 1, exhibiting normalization of the colon's tight junctions. EA at Zusanli acupoint of colon tissue significantly improved the pathological phenotype, showing a therapeutic effect on IBD.

Antidyslipidaemic evaluation of coagulin C and withacoagulin isolated from Withania coagulans (Stocks) dunal berries in hypertension induced secondary dyslipidemia in albino mice

Vascular diseases like hypertension has often been associated with dyslipidemia. Sustained elevated blood pressure for not less than four weeks develops gross irregularities inside arteriolar lumen. The pathological changes might be due to intimal arteriosclerotic thickening causing tapering of the structural narrowing along the arterioles. It might also be due to occurrence of irregular damage in the vascular structure from focal arteriolar constriction and lead to secondary dyslipidemia. The pharmacological intervention may lead to rectification and therapeutic benefit against dyslipidemia.The withanolides coagulin C and withacoagulin were isolated by flash chromatography from the extracts of Withania coagulans (Stocks) Dunal berries. The withanolides were characterized using 1H NMR and 13C NMR, UV and mass spectrometry and confirmed as coagulin C and withacoagulin. The oral toxicity of coagulin C and withacoagulin were investigated in albino mice followed by their antidyslipidemic activity with two dose levels i.e. 25 mg/kg and 50 mg/Kg p.o. This dyslipidemia was secondary to hypertension which was developed by DOCA salt model (25 mg/kg; s.c.+ 2% NaCl). Mice were administered dose of 10, 50 and 500 mg/kg coagulin C and withacoagulin orally for 28 days daily for sub-acute toxicity effect.Coagulin C and withacoagulin were found to be tolerated upto 2000 mg/kg p.o. during acute toxicity study in mice. Coagulin C and withacoagulin were claimed to possess No-Observed Adverse Effect Level (NOEL) at the dose of 500 mg/kg orally. The high dose (50 mg/Kg) withacoagulin has highly significant difference in total cholesterol, triglyceride, HDL, LDL and VLDL levels in comparison to disease control group along with highly significant difference in triglyceride and VLDL level compared to animals of vehicle control group. Coagulin C at both the low (25 mg/Kg) and high dose (50 mg/Kg) have highly significant difference in comparison to vehicle control, disease control and positive control in controlling total cholesterol, triglyceride, LDL and VLDL levels in the experimental animals indicating that coagulin C is not much as effective as withacoagulin in the normalization of altered lipid profile of the experimental animals.On the basis of spectroscopic analysis the isolated compounds from Withania coagulans (Stocks) Dunal berries were confirmed to be coagulin C and withacoagulin. Acute oral toxicity test in mice was found to have safe profile even at high dose for both the compounds. On development of secondary dyslipidemia due to DOCA salt induced hypertension in albino mice, both the isolated compounds showed different degree of correction in the altered lipid profile of the animals. Whereas withacoagulin has been found to have more significant and successful in controlling the altered lipid profile, coagulin C has only marginal effect. The docking studies also confirm the result and hence it can be suggested that withacoagulin is better than coagulin C in managing the alteration of lipid profile of the experimental mice.

Influence of enzyme drug and adsorbent on digestive metabolism processes of steers fattening in technogenic zone

In the practice of feeding of young ruminants for fattening, feed drugs of new generation adsorbents are increasingly and widely introduced. They often exhibit synergistic eff ects on metabolic processes with a large set of biologically active additives including enzyme drugs. The purpose of the research was to study the infl uence of the adsorbent drugs toxfi n and MEC celloviridin G20x on the state of rumen metabolism in fattening steers in whose diets there was an excess content of lead, zinc and cadmium salts. It was found that in order to activate the processes of digestive metabolism due to better elimination of heavy metal salts, it is advisable to jointly introduce the adsorbent toxfi n in the amount of 1 kg/t of compound feed and MEC celloviridin G20x in the amount of 70 g/t of compound feed into the composition of the rations of fattening steers. The results of the conducted studies showed that by the end of fattening at the age of 18 months the animals of the 4th experimental group had the highest body weight, which surpassed the steers of the 1st control group by 29.20 kg (p < 0.05) or 6.86 %. Relative to the control analogues the steers of the 4th experimental group showed an activation of growth in the forestomachs of ciliates (producers of cellulases and amylases) by 54.26 % (p < 0.05) and bacteria of the Flavobacterium vitarumen group (producers of proteinases) by 29.36 % (p < 0.05). Due to the combined feeding of the adsorbent drugs toxfi n and MEC celloviridin G20x, the processes of detoxifi cation of heavy metal salts were more effective. Therefore, the fattening steers of the 4th experimental group more actively accumulated volatile fatty acids in the rumen fl uid, significantly (p < 0.05) outperforming the animals of the 1st control group in this indicator by 1.68 mmol/100 ml or by 20.71 %.

Model of Accelerated Aging in CB6F2 Mice Induced by Ionizing Radiation.

A model for accelerated aging in mice was developed: CB6F2 mice aged 39-45 days were exposed to fractionated 4-fold relatively uniform γ-radiation (137Cs, 0.98 Gy/min) at a total dose of 6.8 Gy. Radiation exposure led to delayed active growth, leukopenia, and lymphopenia for over 1 year during the post-radiation period. The death of irradiated males and females occurred significantly earlier than in control group animals. Median lifespans in the experimental group were 35-38% lower than in the control group (p<0.001). Ionizing radiation exposure led to the early development of hair depigmentation, cachexia, and the development of aging-associated diseases. In irradiated mice, oncological pathology constituted 30-35% in the mortality structure, which is twice as often as in the control group. The developed model can be used to study the pathogenesis of accelerated aging under radiation exposure and the search for means of its prevention and treatment.

Investigation of fingolimod-induced lymphocyte sequestration on inflammatory response and neurological damages after cardiac arrest

BackgroundA sepsis-like syndrome is known to occur after cardiac arrest, leading to cerebral infiltration by white blood cells (WBC). We hypothesized that pharmacological sequestration of WBC, and more specifically lymphocytes within lymphoid tissues, could reduce the cerebral infiltration by these inflammatory cells and subsequent acute brain injury in a porcine model of cardiac arrest. Lymphocyte sequestration was induced by the sphingosine-1 phosphate receptors agonist fingolimod.MethodsIn a first set of experiments, anesthetized pigs underwent a sham instrumentation with no cardiac arrest (n = 4). They received an administration of fingolimod (1 mg/kg, i.v.) in order to confirm its effect on WBC. In a second set of experiments, animals randomly received fingolimod or saline two hours prior to an episode of ventricular fibrillation (14 min) with subsequent resuscitation (n = 6 in each group). Neurological injury was assessed 24 h after resuscitation.ResultsIn the first set of experiments, WBC and blood lymphocyte counts were significantly reduced by − 61 ± 10% and − 75 ± 6% two hours after fingolimod administration. In the second set of experiments, blood lymphocyte counts, but not WBC, were also significantly reduced after cardiac arrest in Fingolimod vs Control group. However, most cytokine blood levels were not different among groups, including Interleukin (IL)-1ra, IL-8 or IL-18 blood levels. A difference was only observed for IL-6, which decreased in Fingolimod vs Control (e.g., 5.6 ± 4.8 vs 59.4 ± 20.6 pg/ml at 2 h after cardiac arrest, respectively; p = 0.126). Neurofilament light chain (NFL) blood levels were not different among groups (57 ± 25 vs 84 ± 41 pg/ml in Fingolimod vs Control at 6 h after resuscitation, respectively). After awakening, 3 and 2 animals were prematurely euthanized for ethical reasons due to recurrent seizures in Fingolimod and Control groups, respectively. At Day 1, neurological dysfunction score was not different between groups (87 ± 7 vs 87 ± 5% in Fingolimod vs Control, respectively). Conversely, a decrease in the number of CD3 + cells was observed in the brain of surviving animals in Fingolimod vs Control group (3.10 ± 0.50 vs 7.53 ± 0.57 CD3 + cells/field, respectively; p = 0.0286).ConclusionFingolimod-induced WBC sequestration, and more specifically lymphocytes sequestration, did not improve clinical neurological dysfunction following cardiac arrest although it reduced cerebral infiltration by lymphocytes.

Understanding the Pathogenesis of Endothelial Injury Induced by Bluetongue Virus in Experimentally Infected Sheep

Bluetongue (BT) is an economically important contagious viral disease transmitted by insect vectors of Culicoides species affecting both domestic and wild ruminants caused by Genus Orbivirus of Reoviridae family. Present in vivo study for the first time conducted to understand the role of oxidative stress and acute phase proteins in the pathogenesis of endothelial injury in target blood vessels of BTV-4 infected sheep. Twelve sheep of either sex were divided into two groups of six animals each and infected intradermally at multiple sites in the neck region with 6×105.9 TCID50/ml of BTV-4 and animals of control group were inoculated with uninfected tissue culture fluid. Animals were sacrificed at 3, 7, 15, 28, 60, and 90 days post infection (dpi) and postmortem examination was conducted. Oxidative stress parameters like LPO, SOD, GPO and catalase in the blood vessels, hemolysate and serum were estimated. Oxidative stress parameter like LPO was upregulated and anti-oxidant parameters like SOD and GPO were downregulated in blood vessels homogenates, hemolysate and serum of BTV-4 infected animals than uninfected control. This shows the role of oxidative stress in causing endothelial cell damage during BTV infection. Acute phase proteins like CRP and fibrinogen in serum and plasma were estimated and found upregulated in BTV-4 infected animals, which shows that inflammatory changes during the disease causes vascular damage. Prothrombin time was estimated in plasma of BTV-4 infected animals and was prolonged in BTV infected animals indicating coagulation disorder. Increased oxidative stress and fibrinogen, and prolonged prothrombin time indicated the coagulation disorders induced by BTV infection by causing endothelial cell damage and severe vascular disease results in congestion, haemorrhages and edema in the blood vessels and other target organs in sheep.

Intravesical Instillation of Hyaluronic Acid With Epidermal Growth Factor for Restoring Urothelial Denudation and Alleviating Oxidative Stress in Lipopolysaccharide-Induced Interstitial Cystitis of Rats.

To investigate the efficacy of an intravesical instillation of hyaluronic acid (HA) combined with epidermal growth factor (EGF) for the treatment of interstitial cystitis (IC) using a lipopolysaccharide (LPS)-induced IC animal model. A total of 24 female Sprague-Dawley rats were randomized to 4 groups: sham control, IC, HA, and treatment (HA/ EGF) groups. A polyethylene-50 tube was placed inside the bladder of each animal. IC was induced by twice-weekly instillations of LPS for 3 weeks, which resulted in chronic injury of the urothelium. Animals in the sham control group only received saline instillation. Treatment solutions of HA and HA/EGF were given on days 0, 7, and 14 after IC induction (400 μL of HA in a concentration of 0.4 mg/0.5 mL and 400 μL of NewEpi, a commercialized HA/EGF mixture containing 2 μg of EGF and 0.4 mg of sodium hyaluronate). Animals were sacrificed on day 21 for further examinations. The HA/EGF group showed visible improvement in hematuria with a significant reduction of red blood cells in the urine compared to the HA group. Histological examination revealed that HA/EGF treatment reversed the abnormalities developed in IC, including infiltration of inflammatory cells, irregular re-epithelialization, and fibrotic tissue. Moreover, HA/ EGF significantly reduced the levels of proinflammation cytokines (tumor necrosis factor-α, interleukin [IL]-6, and IL-1β) and substantially lowered the elevated oxidative stress biomarker malondialdehyde, yet restored the levels of antioxidant enzymes glutathione peroxidase and superoxide dismutase, with superior results than HA treatment. Cystometry studies indicated that HA/EGF significantly prolonged intercontraction interval and increased micturition volume. HA/EGF has been demonstrated as a more effective treatment for enhancing the urothelium lining and reducing inflammatory changes to alleviate clinical symptoms associated with IC in rats, compared to HA alone.

INFLUENCE OF FEED ADDITIVE "ENZAKTIVE MIX" ON THE HEMATOLOGICAL PROFILE OF THE BLOOD OF SOWS AND THEIR PIGLETS

The dynamics of the haematological profile changes and the productivity of sows and piglets obtained from them when feeding the feed additive "EnzActive Mix" were described in the article. The feed additive contains yeast of the genus Saccharomyces cerevisiae with an activity of at least 1.5 x 10^10 CFU/g and a six enzymes complex with protease, cellulase, xylanase, ɑ-amylase, β-gluconase, phytase, which are located under the layer of yeast cells. The results of the study showed that the use of "EnzActive Mix" in sows and their piglets’ diet has a stimulating effect on the oxygen transport function of blood. This is evidenced by a higher number of erythrocytes and platelets, a higher concentration of haemoglobin and haematocrit in the blood of sows of the experimental group compared to the control group on 21st day of lactation (p&lt;0.05). Addition of the feed additive "EnzActive Mix" to the diet caused changes in certain forms of leukocytes ratio in the sows’ blood. Thus, during this period, a higher (p&lt;0.05) relative lymphocytes number and a tendency to decrease of segmented neutrophils number in the blood of experimental group sows compared to the control group were found. Similar changes were noted in the haematological parameters study of piglets obtained from these sows. In particular, the leukocytes number in the blood of the experimental group piglets at 5-, 14- and 28-day age was higher than in their peers of the control group. At the same time, on the 5th day of life, the differences relatively to the control group animals were significant (p&lt;0.05). Feeding of the feed additive led to a decrease in the segmented neutrophils relative number in the blood of the experimental group piglets on the 14th and 28th day of life (p&lt;0.05) and an increase in the lymphocytes number (p&lt;0.05) during the indicated periods of the study. The positive effect of the feed additive "EnzActive Mix" on the productivity parameters of the studied animals was established. In particular, the body weight of experimental group piglets at 5-, 14- and 28-day age was 7.8%, 13.7% (p&lt;0.05) and 8.4% (p&lt;0.01) higher, respectively, than that of the control group.

Оценка системы гемостаза кроликов после прижизненного экспериментального моделирования забрюшинной гематомы

The purpose of research is to study the dynamics of the hemostasis system in rabbits after intravital experimental modeling of retroperitoneal hematoma. The object of the study was 50 male rabbits of the Gray Giant breed, from which experimental and control groups of 25 individuals each were formed. Animals of the experimental group under non-inhalation anesthesia underwent intravital modeling of a retroperitoneal hematoma using a method we developed, including taking blood from the ear artery with the addition of an anticoagulant and subsequent injection of autologous blood into the retroperitoneal space. Animals in the control group were subjected to non-inhalational anesthesia only. Intravital experimental modeling of retroperitoneal hematoma in rabbits led to a number of significant changes in the hemostatic system, which were most pronounced in the first 14 days after the experiment. The international norma-lized ratio (INR) in the experimental group of animals had significantly low values compared to the control group only on the 1st, 3rd and 7th days, and the activated partial thromboplastin time (aPTT) – on the 1st, 3rd days , 7th and 14th days after the experiment. There were no significant differences in the level of fibrinogen in blood plasma between the animals of the experimental and control groups. Thrombin time (TT) in animals of the experimental group was significantly higher only on the 1st, 3rd, 7th and 14th days, and the D-dimer content was significantly higher in all studied periods after the experiment in comparison with the control group animals. The highest level of D-dimer in the blood plasma of animals in the experimental group was recorded on the 7th day and probably indicated that in retroperitoneal hematomas the most active processes of fibrinolysis occur at this time. Low values of INR in the first 7 days and aPTT in the first 14 days, as well as prolongation of TT in the first 14 days after the experiment, give reason to believe that with retroperitoneal hematomas, the threshold for the likelihood of thrombus formation increases during these periods.

L-citrulline Mitigates Systemic Inflammation and Associated Pulmonary Vascular Remodeling in a Neonatal Rat Model of Postnatal Undernutrition

Background &amp; Aim: Preterm infants with postnatal growth restriction and undernutrition are at increased risk of developing bronchopulmonary dysplasia (BPD), pulmonary hypertension (PH), metabolic syndrome, and sepsis. Preclinical and clinical studies report increased systemic inflammation, and lung and pancreatic histopathology in growth-restricted neonates. Safe therapies to mitigate these adverse outcomes associated with postnatal growth restriction and undernutrition in preterm infants are needed. L-citrulline is a non-essential amino acid, involved in the urea and nitric oxide cycles, that exhibits anti-inflammatory properties. This study aims to investigate whether L-citrulline may decrease systemic inflammation and improve histopathology in the lung, heart, and pancreas in a neonatal model of postnatal undernutrition. Methods: Neonatal Sprague Dawley rat pups were divided into 2 groups, normal nutrition (litter size n=12) and undernutrition (litter size n=18) by altering the litter size of the nursing dam. Body weight was measured daily and both groups received daily intraperitoneal injections of saline or L-citrulline (2.5g/kg) for 21 days. On postnatal day 21, heart, lung, and pancreatic tissue were collected for histology and mRNA analyses. Lung and pancreatic tissue were processed for qPCR to detect mRNA expression of pro-inflammatory mediators (MCP-1/CCL2, MIP-1α /CCL3, IL-6, and TNF-α). Right ventricular hypertrophy was calculated using Fulton’s index (weight of the right ventricle divided by the combined weight of the left ventricle and the intraventricular septum). Lung sections were stained with Movat Pentachrome and examined for the presence of immune cells and medial wall thickening of pulmonary arteries. In a separate group of animals, bronchoalveolar lavage fluid (BALF) and blood obtained by cardiac puncture were cytocentrifuged and stained with hematoxylin &amp; eosin for immune cell counts. Results: Undernourished rats had a lower body weight than normally nourished rats at postnatal day 21 (n=11-17; p&lt;0.0001). Undernourished animals had an increased number of immune cells in BALF and lung tissue and an elevated neutrophil count in the blood which were all decreased with concurrent L-citrulline treatment (n=3-4; p&lt;0.001). L-citrulline treatment also prevented the increased mRNA expression of pro-inflammatory mediators in the lungs [MCP-1/CCL2, TNF-α, and IL-1β] (n=8; p&lt;0.05) and pancreas [MCP-1/CCL2, MIP-1α /CCL3, IL-6, and TNF-α] (n=3; p&lt;0.05) of undernourished animals. These animals had a greater pulmonary artery medial wall area and Fulton’s index when compared to normally nourished animals treated with saline or L-citrulline (n=3-4; p&lt;0.001). Concurrent L-citrulline treatment in undernourished animals prevented pulmonary artery vascular remodeling and heart tissue histopathology such that the medial wall area and Fulton’s index did not differ from that of animals in control groups (n=4; p&lt;0.01). Conclusion: L-citrulline effectively mitigates the adverse effects of postnatal undernutrition, reducing lung, pancreatic, and systemic inflammation and histopathology in the lung and heart associated with PH. As a safe, well-tolerated, and inexpensive treatment, L-citrulline presents promising therapeutic potential that could prevent short and long-term morbidities associated with growth restriction and undernutrition in preterm infants. This work was funded by the Women's Auxiliary of The Hospital for Sick Children. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Investigation of Some Ion Channel Expressions in Cochlear Nucleus of Tinnitus Induced Rats

Aim: The aim of this study is to gain a better understanding of how certain ion channels play a role in the molecular mechanisms of salicylate- and noise-induced tinnitus. Method: The present study was conducted on thirty-two, 4-month-old, male Wistar Albino rats. Rats were equally divided into four groups; two experimental groups and two control groups. The assessment of tinnitus was based on a behavioral test which was modified from the conditional suppression method. Tinnitus was induced by sodium salicylate administration and noise exposure in rats in which the suppression ratios were zero (0). All animals in both experimental and control groups were decapitated in deep anaesthesia for 2 h after salicylate or saline administration and noise exposure, consecutively. Tissues from the left and right cochlear nucleus were dissected immediately in ice-cold RNA later (Invitrogen). Before reverse transcription, the RNA pools were arranged. Quantitative changes in HCN1, HCN2, HCN4, SCN1A, SCN2A1, SCN3A, TRPM2, TRPM7 and GAPDH mRNA expressions in the cochlear nucleus in both experimental and control groups were examined by quantitative real-time PCR method. Statistical data were analysed using the SPSS 21 program (Version 21.0, SPSS Inc., Chicago, IL, USA) with the Kruskal-Wallis and Mann-Whitney U tests. Results: Fold changes in the expression levels of SCNA1, SCN2A1, SCN3A, TRPM2, TRPM7, CACNA1B, HCN1, HCN2 and HCN4 genes in both salicylate-induces tinnitus (SAT) and noise-induced tinnitus (NT) groups compared with the control group. According to these data, it is seen that the mRNA levels of all genes are lower in the cochlear nucleus area of the rats in both SAT and NT groups than in the control group. Considering each of these genes in NT group: SCNA1, SCN3A, TRPM7 genes slightly decreased; SCN2A1, TRPM2, HCN1 and HCN4 genes slightly increased compared with the SAT group. For HCN2 gene fold changes were nearly the same in the NT and SAT groups. Conclusion: The findings of this study suggest that tinnitus generation may be closely related to alterations in several key ion channel families activity including voltage-gated calcium channels, hyperpolarization-activated cyclic nucleotide–gated (HCN) channels, transient receptor potential (TRP) channels, voltage-gated sodium channels within the CN, specifically in response to salicylate-induced and noise-induced tinnitus models.

Feed costs and indicators of reproductive capacity of high-yielding cows at different levels of Bypass soybeans in rations

The article presents the results of a scientific and economic experiment to study the influence of different levels of Bypass soybeans in the diets of high-yielding cows on feed costs per kilogram of milk and reproductive functions of cows. Provision of the rations of the experimental cows with an unequal amount of the difficult-to-dissolve fraction of crude protein, at the expense of Bypass soybeans, affected their consumption of bulk feed. The experimental cows consumed 4.1–4.8 kg from the specified 5 kg of alfalfa hay, 20.8–24.6 kg from 25 kg of corn silage, and 8.4–9.7 kg from 10 kg of alfalfa hay. The consumed feed provided cows with crude protein at the level of 16.0–16.5 % of the dry matter, its poorly soluble fraction at the level of 27.3 % of the crude protein in the 1st control group, 31.1 % in the 2nd experimental group, 35.2 % – in the 3rd experimental group and 40.0 % – in the 4th experimental group. The sugar-starch-protein ratio was normal and ranged from 2.20 to 2.28 : 1 with a norm of 2.1 : 1. The micronutrient rations were balanced due to micronutrients of organic origin. On the basis of the obtained data, it was proved that the lowest feed consumption per 1 kg of milk was observed in cows of the 4th experimental group, which were fed rations with 2 kg of Bypass soybeans, where the level of insoluble protein was 40 % of crude protein and these consumptions were 6.5 MJ against 7.4 MJ in the 1st control group. Cows of the 4th experimental group also had the best insemination and service period indicators. The duration of the service period in the experimental groups was less in percentage compared to the animals of the 1st control group by: 15.0 in the 2nd; 25.6 in the 3rd and 32.9 in the 4th. Partial and complete replacement in the diets of cows of the 2nd, 3rd and 4th experimental groups of soybean meal with Vypass soybeans at the level of 1.0; 1.7 and 2 kg of compound feed concentrate in the first 100 days of lactation has a positive effect on the physiological state, stimulates appetite and provides normal conditions for fertilization and embryo development.

Морфофункциональная характеристика печени и поджелудочной железы при воздействии гепатопротектора Ремаксол на фоне острого алкогольного воздействия у крыс

Purpose of the study. To study the effect of hepatoprotective drug Remaxol on morphology of the liver and pancreas as well as on biochemical indices of blood serum in rats with alcohol damage. Material and methods. The study was carried out on sexually mature male white laboratory rats divided into three groups: 1st control group – intact animals (2 rats); 2nd – experimental animals (5 rats), on which acute pancreatitis on the background of alcohol poisoning was modeled by daily administration of 40% aqueous ethanol solution at a dose of 5 mg/kg weight for 7 days without treatment; 3rd – experimental group (5 rats) with alcohol exposure and with administration of Remaxol at a dose of 5 mg/kg weight daily for another 7 days. Morphological study of the liver, its macrostructure was carried out, diameters of hepatocytes and pancreatic acinuses in animals of experimental and control groups were measured, biochemical parameters were estimated. Results. Blood tests indicate acute alcohol poisoning in the experimental groups; short-term (7 days) alcohol damage has less effect on the size of hepatocytes than on the size of pancreatic acini; pancreatic acini were more reactive to the toxic effect of alcohol; against the background of insignificant morphometric differences, the morphological signs of cells and organs changed; in the liver and pancreas there were signs of inflammatory (swelling of cells in the liver and pancreas). Conclusion. Histological picture of the liver and pancreas of animals with acute alcohol damage against the background of Remaxol treatment showed a decrease in the toxic effect of 40% alcohol on hepatocytes and pancreatocytes, normalization of their morphological parameters.

Effect of photobiomodulation therapy as a multimodal approach to treatment of dermal abrasions in cownose rays (Rhinoptera bonasus).

Photobiomodulation therapy (PBMT) has become increasingly utilized in the zoo and aquarium fields to treat ailments in various species; however, its use in teleosts and elasmobranchs is relatively underrepresented in the literature. This study examined the efficacy of PBMT as an adjunct treatment to reduce the healing time of dermal abrasions in cownose rays (Rhinoptera bonasus). Ten cownose rays were included in this study and separated into control (n = 5) and treatment groups (n = 5). Animals in both treatment and control groups received intramuscular injections of enrofloxacin every 72 h to provide broad-spectrum antibiotic coverage. Cownose rays in the treatment group additionally received PBMT using the Companion® CTC-12 unit using the large, noncontact head. Control group rays were subjected to identical handling without the use of PBMT. A total of 196 Joules were delivered at a power of 2 W based on a dose of 5 J/cm2 at each session. Observational analysis revealed a lack of appreciable difference in healing time between the treatment and control groups in the aforementioned settings; however, it is recommended that PBMT protocols should be re-evaluated if there is little to no response in healing after three to four sessions. PBMT use on acute superficial dermal abrasions in cownose rays was well-tolerated in this study and may have utility in a multimodal treatment approach to wound healing. This study also produced a wound scoring metric based on photographs captured at each treatment. Future studies should utilize increased fluence (J/cm2) and irradiance (W/cm2) and incorporate synchronous histological analysis and increased sample size.

Morphological changes in the liver of bulls after the use of adaptogens

The paper presents the results of a study of the influence of adaptogens of plant and animal nature on the productive qualities of Kazakh white-headed bulls and morphological changes in their liver. 40 6-month-old beef bulls were selected for the study, which were divided into 4 groups: I (control) group of bulls consumed only the basic diet, II — adaptogen leucea safflower, III — drone homogenate, IV — pantocrine at the rate of 0.01 ml per 1 kg of animal body weight. At 18 months, a control slaughter of 3 animals from each group was carried out. During the experiment, it was found that the mass of the paired carcass in the bulls of the experimental groups increased by 11.1–21.7 kg (4.25–8.31%, р ≤ 0.05), the slaughter weight by 11.7–23.0 kg (4.24–8.34%, р ≤ 0.05–0.01), the slaughter yield by 0.9–1.9%. The analysis of the morphological composition of the carcass indicates an increase in the specific weight of its edible part of the carcass: by weight of muscle tissue by 4.0–7.7 kg (4.66–8.97%, р ≤ 0.05–0.01), fat — by 0.7–1.4 kg (4.67–9.33%, р ≤ 0.05). There is a high degree of maturity and high nutritional value of the meat of bulls of the experimental groups. Histological studies of the liver of bull calves indicate that when using tincture from drones, the liver structure corresponds to the norm, while in the control group animals some signs of dystrophic changes without pronounced infiltrations and signs of stagnation were found.

Abstract 5002: Next-generation mRNA-LNP vaccine prototype achieves tumor clearance in a GBM mouse model

Abstract Glioblastoma (GBM) is the most prevalent and aggressive primary brain tumor type with an abysmal prognosis, featuring an immunologically dampened tumor microenvironment and limited neoantigen production. Novel therapeutic strategies are urgently needed. A well-characterized GBM-associated mutation that can elicit an anti-tumor immune response is EGFRvIII mutation. The cancer-driver mutation is detected in approximately 30% of patients at the time of diagnosis and plays a pivotal role in the emergence of GBM. EGFRvIII is an in-frame epidermal growth factor receptor (EGFR) deletion generating a constitutively active oncogenic protein. Its highly immunogenic nature represents an ideal focal point for cutting-edge mRNA-LNP candidate therapeutics. To evaluate proprietary lipid nanoparticle (LNP) compositions for their capacity to induce anti-GBM immunity, we developed a murine transplantable GBM tumor model by neonatal electroporation. The somatically engineered glioblastomas harbor two targeted CRISPR knockouts and stably overexpress EGFRvIII to yield transplantable cell lines for orthotopic tumor initiation. One week after intracranial tumor cell implantation of 6-8 week-old female mice (day 0; n=10), half the animals were administered either buffer (control cohort) or our novel anti-EGFRvIII mRNA-LNP vaccine (experimental cohort). The regimen involved four intramuscular injections before monitoring tumor progression by MRI on day 30. The imaging revealed a high tumor burden in all control group animals, compared to no detectable tumor cells in vaccine-protected mice. All control group animals were humanely euthanized by day 38 due to symptoms associated with the growing brain tumor mass. The asymptomatic experimental animals received a fifth anti-EGFRvIII vaccination before MRI-based examinations on day 43 and day 78 confirmed that no tumor cells could be detected in vaccine-protected mice. The results highlight the potency and potential of our proprietary anti-EGFRvIII mRNA-LNP vaccine as a novel anticancer therapeutic. In addition, the established tumor model represents a valuable tool to test future mRNA-LNP designs. Outstanding cellular and molecular investigations will reveal vaccine-mediated mechanistic insights, allowing for the co-advancement of the tumor model and next-generation multi-target mRNA-LNP vaccine candidates. Citation Format: Robert Nechanitzky, Shannon Snelling Snelling, Kristofor Ellestad, Xueqing Lun, Kirstin Olsen, Yu Wu, Yury Karpov, Jun Liu, Matthew Gold, Haritha Menon, Rajesh Krishnan, Robert Georgantas, Pamela S. Ohashi, Douglas J. Mahoney, Jennifer A. Chan, Natalia Martin-Orozco. Next-generation mRNA-LNP vaccine prototype achieves tumor clearance in a GBM mouse model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5002.