Total Nitric Oxide Synthase Activity Research Articles

Strain-specific responses of Penaeus vannamei to two Enterococcus faecium strains supplementation: A trade-off between growth and immunity

Probiotic properties are highly species- or even strain- specific. Strain-specific responses of growth performance, non-specific immunity, disease resistance, and intestinal microbiota in Penaeus vannamei to two Enterococcus faecium strains supplementation were investigated in this study. Basal diet (control, CON) was supplemented with E. faecium HK and LYB at the dose of 1 × 1010 CFU / kg (HK group) and 1 × 1011 CFU / kg (LYB group), respectively. The results indicated that the HK and LYB groups exhibited significant enhancements in growth performance, non-specific immunity, and disease resistance of shrimp against Vibrio parahaemolyticus compared to the control (P < 0.05). However, the specific growth rate of shrimp in the LYB group was significantly better than that in the HK group (P < 0.05). Furthermore, the HK group showed significant increases in phenoloxidase, total nitric oxide synthase activities, lysozyme content in the serum, as well as the relative expression levels of proPO, LGBP, Imd, Relish, and eIF4E2 genes in the hepatopancreas compared to the LYB group (P < 0.05). These results suggested the strain HK exhibited better non-specific immunity efficiency while the strain LYB exerted better growth enhancement in shrimp. Dietary both of strain HK and LYB significantly altered microbial community structure (P < 0.05), and the strain HK significantly improved intestinal microbial diversity in shrimp. The relative abundance of Rubritaleaceae and Rhodobacteraceae were significantly increased, whereas those of facultative anaerobes and potential pathogens (i.e., Vibrionaceae, Flavobacteriaceae and Erysipelotrichaceae, etc.) were reduced in response to dietary supplementation of E. faecium HK and LYB. Furthermore, dietary both of strain HK and LYB could promote intestinal microbial community stability by increasing average connectivity, cooperation interactions, the number of generalists within the network. When growth performance and intestinal microbial homeostasis of shrimp are considered, E. faecium LYB might be better than the strain HK in shrimp farming.

Вплив модуляторів факторів транскрипції NF-κB і Nrf2 на показники оксидативно–нітрозативного стресу в скелетних м’язах щурів за умов хронічної гіпомелатонінемії та вуглеводно-ліпідної дієти

This article describes the effect of NF-κB inhibitor ammonium pyrrolidine dithiocarbamate and Nrf2 inducer dimethylfumarate on indicators of oxidative-nitrosative stress in skeletal muscles of rats with chronic hypomelatoninemia, induced by roundthe-clock illumination with an intensity of 1500 lux for 30 days, against the background of a carbohydrate-lipid diet (20% fructose solution and appropriate food) for 60 days. The study demonstrated that the administration of ammonium pyrrolidine dithiocarbamate and dimethylfumarate under the experimental conditions impedes the development of hypomelatoninemia, this was accompanied by a two-fold increase in the blood serum melatonin concentration compared to the control group. Moreover, the administration of ammonium pyrrolidine dithiocarbamate and dimethylfumarate significantly reduces the production of superoxide anion radical by electron transport chains of mitochondria (by 47.9 and 51.3%) and sarcoplasmic reticulum (by 48.6 and 52.0%) in the homogenate of femoral muscles, the total activity of NO synthase (by 37.2 and 36.2%) and its inducible isoform (by 41.1 and 40.0%), the concentration of peroxynitrites of alkaline and alkaline earth metals (by 37.2 and 41.0 %), while the activity of constitutive NO-synthases (by 2.9 and 2.8 times) and their conjugation index (by 5.2 and 5.4 times) increases compared to the respective control values. We suggest that modulators of redox-sensitive transcription factors NF-κB and Nrf2 under conditions of chronic hypomelatoninemia and the exposure to the carbohydrate-lipid diet are effective means to restrain the development of oxidative-nitrosative stress in skeletal muscles.

Photobiomodulation therapy protects red blood cells against nitrative stress during streptozotocin-induced diabetes

Background. According to the International Diabetes Federation Diabetes Atlas, 10th edition, diabetes is responsible for 6.7 million deaths in 2021. Diabetes mellitus is a group of metabolic disorders characterized by hyperglycemia secondary to either resistance to insulin, insufficient insulin secretion, or both. Oxidative and nitrative stress is a vital part of the complex mechanism by which diabetes and its complications develop. It is known that Photobiomodulation therapy accelerates diabetic wound healing, treats relegated inflammation, and increases oxygen availability for cells. Although some basic molecular mechanisms caused by photobiomodulation therapy in different cell types are already known, they have not been studied in erythrocytes and are different due to the absence of central organelles such as nucleus and mitochondria. The aim of the study was to investigate the effect of photobiomodulation therapy on the development of nitrative stress in blood plasma and erythrocytes of rats from different experimental groups. Materials and Methods. The study was performed on white outbred male rats weighing 130–180 g. The diabetes mellitus was induced by intraperitoneal injection of streptozotocin (60 mg/kg). Rats were exposed to photobiomodulation with light-emitting diodes at a wavelength of 630–660 nm daily for 10 days. The irradiation time was 5 minu­tes. The content of nitrite and nitrate anions, total NO synthase activity, as well as the activity of its endothelial and inducible isoforms in red blood cells of rats were determined spectrophotometrically. Results and Discussion. Under streptozotocin-induced diabetes mellitus, the content of nitrite and nitrate anions and NO synthase activity increased in the rats’ red blood cells, as well as in blood plasma. Moreover, we found an increase in inducible NO synthase activity and nitrate ion content in red blood cells of irradiated healthy rats. Also, there was an increase in nitrite and nitrate ion content after photobiomodulation therapy in the blood plasma of healthy animals. On the other hand, irradiation caused a decrease in NO synthase activity with a parallel reduction in both nitrite and nitrate anions content in erythrocytes and blood plasma of rats with experimental diabetes. Conclusion. Photobiomodulation therapy protects rats’ red blood cells from nitrative stress during streptozotocin-induced diabetes mellitus.

Positive effects of replacing commercial feeds by fresh black soldier fly (Hermetia illucens) larvae in the diets of Pacific white shrimp (Litopenaeus vannamei): Immune enzyme, water quality, and intestinal microbiota

This study investigated the effects of replacing commercial feeds with fresh black soldier fly larvae (BSFL) on the immune enzyme activities, water quality in the culturing environment, and intestinal microbiota of Pacific white shrimp (Litopenaeus vannamei). Five different feeding regimes were designed, in which 0% (control group), 25%, 50%, 75%, and 100% commercial feeds were replaced with equal wet weight of fresh BSFL, respectively. The experiment lasted for 45 days, and the results showed that the serum T-NOS (total nitric oxide synthase), AKP (alkaline phosphatase), and ACP (acid phosphatase) activity were significantly increased in the BSFL25% group. However, the immune enzymes of the hepatopancreas decreased significantly in all the BSFL-containing groups compared to the control group. Total ammonia nitrogen concentrations throughout the experimental phase and nitrate concentrations in the middle and later phases of the experiment were negatively correlated with the replacement rate of fresh BSFL in the diet, whereas pH was positively correlated with the replacement rate of fresh BSFL in the diet. Proteobacteria and Bacteroidetes were dominant phyla in the intestines of shrimps, but the relative abundances of Proteobacteria and Firmicutes decreased, with those of Bacteroidetes and Planctomycetes increased in the BSFL-containing groups. Among them, higher relative abundances of potential probiotics such as Motilimonas, Shimia, Pseudoalteromonas, and Shewanella and lower relative abundance of genus Vibrio were observed in the fresh BSFL-containing groups. Furthermore, shrimps fed with BSFL-containing diets had higher bacterial richness and diversity in the intestines. In conclusion, a proper replacement of commercial feed with fresh BSFL had a positive effects on the immune-related enzyme activities, water quality, and intestinal health in the L. vannamei.

Effects of Dietary Supplementation with Glutamine on the Immunity and Intestinal Barrier Gene Expression in Broiler Chickens Infected with Salmonella Enteritidis.

Simple SummarySalmonella Enteritidis, a Gram-negative bacterium, is ingested into the gut of animals, especially young chicks. Glutamine (Gln), a nonessential amino acid, is an important energy source and active free radical-scavenging compound for enterocyte and lymphocyte cells. In this study, we found that Gln might alleviate the intestinal inflammatory response and increase the intestinal immune barrier function in broilers challenged with Salmonella Enteritidis. During the experimental period, supplementation with Gln improved growth performance, increased serum immunoglobulin concentrations and intestinal mucosa Bcl-2 mRNA expression levels, and decreased the lysozyme (LZM, only serum) and nitric oxide (NO) activities in serum and the intestinal mucosa, along with the intestinal mucosa barrier gene mRNA expression levels. These findings provide useful information regarding the intestinal immune barrier function of broilers.The effects of glutamine (Gln) on immunity and intestinal barrier gene expression levels in broilers challenged with Salmonella Enteritidis were evaluated. A total of 400 1-day-old broilers were randomly assigned to four groups, 10 repetition treatments per group with 10 broiler chickens for a 21-day feeding trial. The groups were the normal control group (CON, no infected group, fed with a basal diet); the S. Enteritidis-infected control group (SCC, infected with 2.0 × 104 CFU/mL of S. Enteritidis, fed a basal diet); and the Gln 1 and 2 groups, who were challenged with S. Enteritidis and fed a basal diet plus Gln at 0.5% and 1.0%, respectively. The results show that S. Enteritidis had adverse effects on the average daily feed intake, average daily gain, and the feed conversion ratio of infected broilers compared with those of CON broilers on d 7 (p < 0.05); decreased serum immunoglobulin A (IgA), immunoglobulin M (IgM), and immunoglobulin G (IgG) concentrations, and intestinal mucosa Bcl-2 mRNA expression levels (p < 0.05); increased the Lysozyme (LZM, only serum), NO, inducible NO synthase (iNOS) (except at 4 d), and total nitric oxide synthase (TNOS) (except at 4 d) activities in serum and the intestinal mucosa; and increased intestinal mucosa polymeric immunoglobulin receptor (pIgR) (except at 21 d), Avian beta-defensin 5 (AvBD5), AvBD14, Bax, and Bak mRNA expression levels during the experimental period (p < 0.05). Supplementation with Gln improved growth performance; increased serum IgA, IgG, and IgM concentrations and intestinal mucosa Bcl-2 mRNA expression levels (p < 0.05); decreased the LZM (only serum), NO, iNOS (except at 4 d), and TNOS (except at 4 d) activities in serum and the intestinal mucosa; and decreased intestinal mucosa pIgR (except at 21 d), AvBD5, AvBD14, Bax, and Bak mRNA expression levels during the experimental period (p < 0.05). These results suggest that Gln might lessen the inflammatory reaction of the small intestine and enlarge the small bowel mucosa immune and barrier function in broiler chickens challenged with S. Enteritidis.

Beneficial Changes in Growth Performance, Antioxidant Capacity, Immune Response, Hepatic Health, and Flesh Quality of Trachinotus ovatus Fed With Oedocladium carolinianum

The purpose of this study is to assess the feasibility of astaxanthin-rich Oedocladium carolinianum as an immunostimulant in the diet for Trachinotus ovatus. Three experimental diets containing 0% (OC0), 1% (OC1), and 5% (OC5) O. carolinianum powder were formulated for 6-week feeding trials. The results indicated that the OC5 diet boosted the growth performance through decreasing the feed conversion ratio and increasing digestive enzyme activities and intestinal villus length. Meanwhile, fish fed with the OC5 diet promoted antioxidant ability via stimulating the Nrf2-ARE signal pathway and enhancing antioxidant enzyme activities. Furthermore, the OC5 diet exerted hepatoprotective effects by suppressing the lipid deposition and inflammation response and enhancing the transport capacity of cholesterol. Besides, the OC5 diet improved the non-specific immunity by activating the lysozyme and complement system and increasing the nitric oxide content and total nitric oxide synthase activity. Dietary O. carolinianum supplementation promoted the deposition of astaxanthin in the whole body. Therefore, a diet supplemented with 5% O. carolinianum is recommended to boost the growth, antioxidant capacity, immune response, and flesh quality of T. ovatus.

Effects of Barranca yajiagengensis Powder in the Diet of Trachinotus ovatus on the Growth Performance, Antioxidant Capacity, Immunity and Morphology of the Liver and Intestine.

Barranca yajiagengensis, a novel filamentous microalga, can accumulate lutein under high-light and low-nitrogen conditions. It is well known that lutein has antioxidant, anti-inflammatory and immune-modulating properties. The purpose of this study is to evaluate the effects of including lutein-rich B. yajiagengensis powder in the diet of Trachinotus ovatus on the growth performance, antioxidant capacity, immunity, liver, and intestinal morphology. For this aim, three experimental diets containing 0% (BY0), 1% (BY1), and 5% (BY5) B. yajiagengensis powder were formulated for six-week feeding trials. The results indicated that growth performance, feed utilization, and intestinal morphology were not affected by different diet treatments. Fish fed with the BY5 diet promoted antioxidant ability by activating the Nrf2-ARE signal pathway and enhancing antioxidant enzymes activities. Furthermore, the BY5 diet improved non-specific immunity and antibacterial ability by activating lysozymes and the complement system and increasing the nitric oxide (NO) content and total nitric oxide synthase activity. Dietary B. yajiagengensis supplementation improved the liver morphology and exerted hepatoprotective effects. Therefore, as a natural source of lutein, B. yajiagengensis has the potential as a safe and non-toxic immunostimulant for T. ovatus. A diet supplemented with 5% B. yajiagengensis is recommended to improve the growth, antioxidant capacity, immune response, and liver health of T. ovatus.

Effects of Water Extract from Artemisia argyi Leaves on LPS-Induced Mastitis in Mice.

Simple SummaryMastitis is a common disease in dairy cows. On the one hand, it will reduce milk yield and milk quality of dairy cows, thus increasing the cost of animal husbandry, and, on the other hand, it will influence the health of infected animals and even human beings. Generally speaking, because mastitis is caused by pathogenic microorganisms, antibiotic treatment is commonly used. However, antibiotic resistance of microorganisms caused by wrongful use of antibiotics and antibiotic residues after treatment has become an urgent problem to be solved. Chinese herbal medicines are pure natural substances, and many of them have antibacterial, anti-inflammatory, or immune-enhancing effects. In this experiment, Artemisia argyi (A. argyi) was selected as the research object to construct the cell model of cow mastitis. Studies have found that A. argyi extract can play a positive role in the regulation of inflammation, which is rich in organic acids and flavonoids. Therefore, A. argyi extract may be a potential treatment for mastitis.In the context of the unsatisfactory therapeutic effect of antibiotics, the natural products of plants have become a research hotspot. Artemisia argyi (A. argyi) is known as a traditional medicine in China, and its extracts have been reported to have a variety of active functions, including anti-inflammatory. Therefore, after establishing the mouse mastitis model by lipopolysaccharide (LPS), the effects of A. argyi leaves extract (ALE) were evaluated by pathological morphology of the mammary gland tissue, gene expression, and serum oxidation index. Studies have shown that ALE has a restorative effect on LPS-induced mammary gland lesions and significantly down-regulated the rise of myeloperoxidase (MPO) induced by LPS stimulation. In addition, ALE played a positive role in LPS-induced oxidative imbalance by restoring the activities of glutathione peroxidase (GSH-PX) and superoxide dismutase (SOD) and preventing the increase in nitric oxide (NO) concentration caused by the over-activation of total nitric oxide synthase (T-NOS). Further analysis of gene expression in the mammary gland showed that ALE significantly down-regulated LPS-induced up-regulation of inflammatory factors IL6, TNFα, and IL1β. ALE also regulated the expression of MyD88, a key gene for toll-like receptors (TLRs) signaling, which, in turn, regulated TLR2 and TLR4. The effect of ALE on iNOS expression was similar to the effect of T-NOS activity and NO content, which also played a positive role. The IκB gene is closely related to the NF-κB signaling pathway, and ALE was found to significantly alleviate the LPS-induced increase in IκB. All of these results indicated that ALE may be considered a potential active substance for mastitis.

Effect of FLU-ACIL RHINO nasal spray on the inflammatory process and recovery of patients after FESS

Topicality: The main method of treatment of chronic rhinosinusitis is functional endoscopic rhinosurgery (FESS). However, the regeneration of tissues damaged during surgery remains important. It has three stages – alteration (primary and secondary), exudation and proliferation – and takes place with the formation of reactive oxygen species. Therefore, determination of total NO synthase activity (NOS), inducible synthase activity (iNOS) and constitutive (cNOS) isoforms are biochemical markers of the inflammatory response. The activity of the alteration process and the degree of postoperative damage is characterized by the concentration of glycosaminoglycans and L-oxyproline. Materials and methods: This study determined the multimodal (multi-vector) effect of Flu-Acyl rhino spray (6% N-acetylcysteine in 3% hypertonic pH-controlled saline) on the inflammatory response of the nasal mucosa in the postoperative period. The study was conducted at the Department of Otolaryngology of NMU named after O.O. Bogomolets on the basis of "Alexander Clinical Hospital of Kyiv". Clinical and laboratory examinations of 52 patients aged 18 to 60 years who underwent FESS were performed. Patients were randomized into two groups. 24 patients received standard treatment (control group), 26 patients – with the addition of nasal spray Flu-Acyl rhino (main group). Results: It was found that the indicators of total activity of NO synthase (NOS) and its isoforms, as well as the concentration of glycosaminoglycans and L-oxyproline were lower in patients of the main group on days 7 and 14 of the postoperative period compared with patients in the control group (р&lt;0,05). These results can be explained by the anti-inflammatory effect of Flu-Acyl rhino spray due to the presence of N-acetylcysteine, which is able to inhibit the activation of the transcription factor NF-kB, which leads to decreased iNOS activity on the 7th and 14th day of treatment. N-acetylicysteine also reduces the degradation of amorphous connective tissue, shifts the balance in the proteolysis / antiproteolysis system towards the predominance of the antiproteolytic component, participates in the normalization of physiological production and utilization of sulfide anion. Conclusions: The use of Flu-Acyl rhino nasal spray in the postoperative period due to multimodal action promotes physiological changes from alteration and exudation to proliferation processes, normalizes mucociliary clearance in the postoperative period, improves healing and reduces recovery and disability.

Исследование влияния адемола на обмен оксида азота в головном мозге крыс с черепно-мозговой травмой

Актуальность. Высокие концентрации оксида азота (NO) приводят к прогрессированию нарушений мозгового кровообращения на фоне черепно-мозговой травмы (ЧМТ). Одним из ведущих молекулярных механизмов нейроцитопротекторного действия современных фармакологических препаратов является их корректирующее воздействие на обмен NO. Цель: оценить влияние адемола по сравнению с амантадином сульфатом на состояние L-аргинин/NO в головном мозге крыс с ЧМТ. Материалы и методы. Опыты проведены на крысах-самцах. Экспериментальную модель тяжелой ЧМТ создавали с использованием пневматического пистолета. Терапевтическое действие адемола при ЧМТ оценивали в дозе 2 мг/кг с интервалом 2 р/сутки в течение 8 суток. В качестве лекарственного средства для контрольной группы применяли 0,9% NаСl в дозе 2 мл/кг, а препарата сравнения — амантадина сульфат в дозе 5 мг/кг. Для определения эффективности исследуемых препаратов использовали уровень L-аргинина и суммарную активность NO-синтазы. Результаты. Сравнительный анализ эффективности адемола и амантадина сульфата на 8-е сутки наблюдения показал, что в группе животных с ЧМТ, которым применяли адемол, содержание L-аргинина в мозге было больше на 112 % (р &lt; 0,05), чем в группе контроля. В этих условиях суммарная активность NO-синтазы в головном мозге была меньше на 26,6 % (р &lt; 0,05) относительно нелеченых животных с ЧМТ. Зато у животных, леченных амантадином сульфатом, уровень L-аргинина в головном мозге был больше на 72 % (р &lt; 0,05), чем в группе контрольной патологии. При этом суммарная активность NO-синтазы в головном мозге была меньше на 15,4 % (р &lt; 0,05). Выводы. Экспериментальное моделирование травматического поражения головного мозга сопровождалось развитием в его структурах пертурбаций в системе L-аргинин/NO-синтаза. Одним из патогенетических механизмов защитного действия на головной мозг при ЧМТ является способность адемола предупреждать истощение в поврежденном мозге запасов аминокислоты L-аргинина и гиперактивацию NO-синтазы, причем по указанному эффекту адемол превосходил амантадина сульфат (р &lt; 0,05).

The effect of photobiomodulation therapy of some indices of rats’ blood cells functional state under experimental diabetes mellitus

Background. Diabetes mellitus is a chronic endocrine-metabolic disease caused by an absolute or relative insulin deficiency. During diabetes, there are perfect conditions for the development of oxidative stress: the content of substrates for oxidation increases, the content of natural antioxidants decreases and the activity of antioxidant systems is suppressed. It is known that photobiomodulation therapy produce antioxidant and antihyperglycemic effects. Here we investigated its influence on blood system functioning. Materials and Methods. The study was performed on male Wistar rats. Experimental diabetes mellitus was induced by the intraperitoneal injection of streptozotocin. Leukocyte formula was calculated using blood smears stained by Romanowsky–Giemsa. Catalase activity was determined spectrophotometrically. Affinity of hemoglobin to oxygen was evaluated by spectrophotometric method in Ivanov’s modification by drawing oxygenation curves. The protoporphyrin content in whole blood was measured by analyzing its fluorescence spectra. The content of NO2-, total and inducible NO synthase activity was determined spectrophotometrically. Results. Under the action of photobiomodulation therapy on healthy animals, there was a shift of oxygenation curves to the left and a decrease of P50, whereas under irradiation of rats with diabetes, there was a shift of oxygenation curves to the right and increase in P50 compared to indices in nonirradiated animals. During diabetes, there was a decrease in protoporphyrin content compared to control, but there was a tendency to increase under photobiomodulation. Photobiomodulation therapy of rats with diabetes increased catalase activity in erythrocyte hemolysates. We revealed significant changes in leukocyte formula under photobiomodulation. The total NO synthase activity in leukocytes of rats with diabetes was higher compared to healthy animals, but decreased under the action of photobiomodulation. We found an increase in inducible NO synthase activity in leukocytes of rats with diabetes and in leukocytes of irradiated healthy animals. An increase in NO2- content in leukocytes of rats with diabetes was detected. Under photobiomodulation, NO2- content was significantly lower in rats with diabetes. Conclusion. Photobiomodulation therapy produces a corrective action on blood system during diabetes, in particular, it improves oxygen release from hemoglobin and prevents hypoxia. Simultaneously with the increase in tissue oxygen saturation, a decrease in NO synthase activity and nitrite content along with an increase in catalase activity prevents the development of oxidative stress.

Dietary Supplementation with Enterococcus faecium R1 Attenuates Intestinal and Liver Injury in Piglets Challenged by Lipopolysaccharide.

Simple SummaryThe biological characteristics of E. faecium R1 and the effect of dietary supplementation with E. faecium R1 on the growth performance of weaned piglets were studied. The results showed that E. faecium R1 had the characteristics of effective bacteriostatic activity, acid resistance, bile salt resistance, and high-temperature resistance. Dietary supplementation with E. faecium R1 (6.5 × 106 CFU/g) improved intestinal function of weaning piglets by decreasing diarrhea incidence. Further research found that dietary supplementation with E. faecium R1 (6.5 × 106 CFU/g) attenuated intestinal and liver injury in piglets challenged by lipopolysaccharide.In this study, a strain of E. faecium R1 with effective bacteriostatic activity, acid resistance, bile salt resistance, high-temperature resistance was screened. To study the effect of E. faecium R1 on lipopolysaccharide (LPS)-induced intestinal and liver injury in piglets, twenty-four weaned female piglets were randomly assigned into one of three groups (8 piglets per group). Piglets in the control group and LPS group were fed a basal diet, piglets in the E. faecium group were fed the basal diet supplemented with E. faecium R1 (6.5 × 106 CFU/g). On day 21 of the trial, piglets in the LPS group and E. faecium group were intraperitoneally administered LPS (100 μg/kg), piglets in the control group were administered the same volume of saline. Subsequently, blood samples were collected at 3 h, and intestinal, liver, and pancreas samples were collected at 6 h. Results showed that E. faecium R1 supplementation significantly decreased the diarrhea rate and feed to gain ratio, and dramatically reduced LPS-induced intestinal and liver injury in piglets. Compared with the LPS group, E. faecium R1 supplementation significantly increased the content of glucagon in plasma and IL-1β in the liver, and the mRNA levels of villin in jejunum and ileum and Bcl-xL and pBD-L in the ileum, and significantly decreased the contents of prostaglandin 2 and malondialdehyde in the liver and the activities of myeloperoxidase and aspartate aminotransferase in plasma in piglets. Moreover, E. faecium R1 improved the pancreatic antioxidant capacity in piglets, which was indicated by a significant increase in catalase activity and a decrease in total nitric oxide synthase activity. In summary, dietary supplementation with E. faecium R1 alleviates intestinal and liver injury in LPS-challenged piglets.

Activation of glucocorticoid receptors is associated with the suppression of antioxidant responses in the liver of goats fed a high-concentrate diet

This study investigated changes in oxidative stress and the relevant mechanisms in the liver of goats fed a high-concentrate (HC) diet for 5 weeks. Twelve goats were randomly assigned to a low-concentrate (concentrate-to-forage = 55:45, LC, n = 6) or HC diet (concentrate-to-forage = 90:10, n = 6), with dry matter as the base. Enzyme activity assays, real-time polymerase chain reaction and western blotting were used to evaluate antioxidant parameters and gene expression in the liver. Superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), total nitric oxide synthase activity and total antioxidant capacity (T-AOC) in the liver declined (p < .01) in HC-fed goats compared to those in LC-fed goats. The mRNA levels of GPX1, CAT and SOD1 were down-regulated (3.69, 47.37 and 27.61%, respectively) in HC-fed goats compared to those in LC-fed goats. Furthermore, glutathione S-transferase M1 (GSTM1) was upregulated (466.35%, p < .01) in the liver of HC-fed goats. The mRNA and protein levels of the nuclear factor E2-related factor 2 (NRF2) and total glucocorticoid receptor (GR) declined (p < .05) in HC-fed goats (by 28.57, 33.1, 30.85 and 34%, respectively). However, the nuclear translocation of GR increased (p < .05; by 44.75%) in HC-fed goats. Negative correlations were detected for hepatic nuclear GR protein expression with hepatic CAT activity and GPx activity. In conclusion, feeding an HC diet to goats for 5 weeks suppressed NRF2-dependent antioxidant responses and enhanced GR nuclear translocation in the liver. Highlights Feeding goats high-concentrate diets suppresses NRF2-dependent antioxidant responses in liver, downregulating NRF2 and altering SOD1, GPx and CAT activity. This suppression of NRF2-dependent antioxidant responses is associated with GR activation in the liver of goats.

Chronic L-Name-Treatment Produces Hypertension by Different Mechanisms in Peripheral Tissues and Brain: Role of Central eNOS.

The goal of our study was to analyze the time course of the effect of NG-nitro-L-arginine methyl ester (L-NAME) on nitric oxide synthase (NOS) isoforms and nuclear factor–κB (NF-κB) protein expression, total NOS activity, and blood pressure (BP) in rats. Adult 12-week-old male Wistar rats were subjected to treatment with L-NAME (40 mg/kg/day) for four and seven weeks. BP was increased after 4- and 7-week L-NAME treatments. NOS activity decreased after 4-week-L-NAME treatment; however, the 7-week treatment increased NOS activity in the aorta, heart, and kidney, while it markedly decreased NOS activity in the brainstem, cerebellum, and brain cortex. The 4-week-L-NAME treatment increased eNOS expression in the aorta, heart, and kidney and this increase was amplified after 7 weeks of treatment. In the brain regions, eNOS expression remained unchanged after 4-week L-NAME treatment and prolonged treatment led to a significant decrease of eNOS expression in these tissues. NF-κB expression increased in both peripheral and brain tissues after 4 weeks of treatment and prolongation of treatment decreased the expression in the aorta, heart, and kidney. In conclusion, decreased expression of eNOS in the brain regions after 7-week L-NAME treatment may be responsible for a remarkable decrease of NOS activity in these regions. Since the BP increase persisted after 7 weeks of L-NAME treatment, we hypothesize that central regulation of BP may contribute significantly to L-NAME-induced hypertension.

INOS is not responsible for RyR1\xa0S-nitrosylation in mdx mice with truncated dystrophin

BackgroundPrevious research indicated that nitric oxide synthase (NOS) is the key molecule for S-nitrosylation of ryanodine receptor 1 (RyR1) in DMD model mice (mdx mice) and that both neuronal NOS (nNOS) and inducible NOS (iNOS) might contribute to the reaction because nNOS is mislocalized in the cytoplasm and iNOS expression is higher in mdx mice. We investigated the effect of iNOS on RyR1 S-nitrosylation in mdx mice and whether transgenic expression of truncated dystrophin reduced iNOS expression in mdx mice or not.MethodsThree- to 4-month-old C57BL/6 J, mdx, and transgenic mdx mice expressing exon 45–55-deleted human dystrophin (Tg/mdx mice) were used. We also generated two double mutant mice, mdx iNOS KO and Tg/mdx iNOS KO to reveal the iNOS contribution to RyR1 S-nitrosylation. nNOS and iNOS expression levels in skeletal muscle of these mice were assessed by immunohistochemistry (IHC), qRT-PCR, and Western blotting. Total NOS activity was measured by a citrulline assay. A biotin-switch method was used for detection of RyR1 S-nitrosylation. Statistical differences were assessed by one-way ANOVA with Tukey-Kramer post-hoc analysis.Resultsmdx and mdx iNOS KO mice showed the same level of RyR1 S-nitrosylation. Total NOS activity was not changed in mdx iNOS KO mice compared with mdx mice. iNOS expression was undetectable in Tg/mdx mice expressing exon 45–55-deleted human dystrophin, but the level of RyR1 S-nitrosylation was the same in mdx and Tg/mdx mice.ConclusionSimilar levels of RyR1 S-nitrosylation and total NOS activity in mdx and mdx iNOS KO demonstrated that the proportion of iNOS in total NOS activity was low, even in mdx mice. Exon 45–55-deleted dystrophin reduced the expression level of iNOS, but it did not correct the RyR1 S-nitrosylation. These results indicate that iNOS was not involved in RyR1 S-nitrosylation in mdx and Tg/mdx mice muscles.

Colonization and Probiotic Effect of Metschnikowia sp. C14 in the Intestine of Juvenile Sea Cucumber, Apostichopus japonicus

Viable cell count was used to determine whether Metschnikowia sp. C14 can colonize the intestine of juvenile sea cucumber Apostichopus japonicus . Sea cucumber individuals were divided into two groups, which were fed the control diet for 38 days or the C14-supplemented diet at 10 5 cells g −1 diet for 28 days, then the control diet from day 29 to day 38. The number of C14 cells in the intestine of sea cucumber fed the C14-supplemented diet significantly increased from day 7 to day 28, and decreased from day 29 to day 38. Sea cucumber fed with the diet containing C14 showed a significant increase in trypsin activity and lipase activity from day 21 to day 33 compared with the control. Feeding C14 significantly improved the phagocytic activity and respiratory burst in coelomocytes from day 21 to day 35 and from day 14 to day 38, respectively. In addition, there was an obvious enhancement in lysozyme activity (from day 21 to day 38 or day 33), phenoloxidase activity (from day 21 to day 28) and total nitric oxide synthase activity (from day 14 to day 38) in coelomic fluid supernatant and/or coelomocyte cell lysate supernatant compared with the control. There were significant positive correlations between the number of C14 cells colonizing the intestine and trypsin activity of the intestine, lysozyme activity of the coelomic fluid supernatant and coelomocyte lysate supernatant from sea cucumber. These data suggested that the number of C14 cells should be maintained at 10 5 cfu (colony-forming units) g −1 intestine material for the maximum benefit.

Decreased Level of Neuropeptide Y Is Associated With Gastroparesis in Male Diabetic Rats.

BackgroundSubstance P (SP) and neuropeptide Y (NPY), excitatory and inhibitory neuropeptides, respectively, may impact gastric motility in patients with diabetic mellitus (DM). We investigated these neuropeptide levels, NPY receptors, total nitric oxide synthase (NOS) levels, and neuronal NOS alpha (nNOSα) activation status and levels in streptozotocin-induced type I diabetes in male rats.MethodsRats were grouped based on serum glucose and gastric emptying time: normal untreated control (CM), diabetic (DM) and diabetic gastroparesis (DM + GP). Neuropeptide serum levels were determined by enzyme-linked immunosorbent assay (ELISA). Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting measured NPY receptors, Y1 and Y2, and nNOSα expression. Low-temperature SDS-PAGE followed by western blotting was used to measure the dimerization of nNOSα. An NOS colorimetric assay kit was used to measure total NOS activity.ResultsSP levels were significantly decreased (P < 0.05) in DM and DM + GP compared to CM. NPY levels were significantly decreased (P < 0.05) in DM compared to CM, and DM + GP had a more significantly decreased NPY when compared to both DM and CM. Protein levels of neuropeptide receptor Y1 (NPY-Y1) in the smooth muscle of pylorus were significantly increased in DM, but not in DM + GP when compared to CM. Neuropeptide receptor Y2 (NPY-Y2) was not detected. Changes in nNOSα activity and their protein levels, as well as total NOS activity, among the groups were insignificant.ConclusionsIncreased expression of pylorus NPY-1R and decreased serum NPY are present in diabetes. A more pronounced decreased serum NPY with no NPY-1R upregulation in pyloric smooth muscle is associated with gastroparesis. NPY levels show no relationship with nNOSα levels, their activation status, or total NOS activity in pyloric smooth muscle. These data suggest a pathophysiological role of severely depleted NPY and absence of NPY-Y1 upregulation for gastroparesis phenotype.

EFFECT OF AP-1 TRANSCRIPTION FACTOR INHIBITOR ON STRUCTURAL, METABOLIC AND BIOMECHANICAL CHANGES IN BONE TISSUE DURING COMBINED EXCESSIVE INTAKE OF SODIUM FLUORIDE AND SODIUM NITRATE

This article highlights the effect produced by the inhibitor of the AP-1 transcription factor activation on the mechanisms of structural, metabolic and biomechanical disorders in the femoral bones and vertebrae during combined excessive intake of sodium fluoride and sodium nitrate. The experiment was conducted on 30 white rats divided into 4 groups: the 1st included the intact animals, the 2nd group involved the rats subjected to the co- administration of sodium fluoride (10 mg / kg body weight) and sodium nitrate (500 mg / kg body weight) for 30 days, the 3rd group included the animals, which starting from the 15th day of intoxication, were injected SR 11302 ((E, E, Z, E) -3-Methyl-7- (4-methylphenyl) - 9- (2,6,6-trimethyl-1-cyclohexen-1-yl) -2,4,6,8-nonatetraenoic acid), an inhibitor of AP-1 activation in a dose of 1 mg / kg intraperitoneally 3 times a week. It has been revealed that the SR 11302 administration restores the mechanism of NO autoregulation in the femoral bones during the sodium fluoride and sodium nitrate co- administration, reducing the total activity of NO synthase and activity of its inducible isoform under a reciprocal increase in total arginase activity, and suppresses the peroxynitrite production. This is accompanied by a decrease in the activity of enzymes, which are known as markers of bone resorption (acid phosphatase and its bone isoform) and restriction of the depolymerization of collagen, proteoglycans and sialoglycoproteins of the connective (bone) tissue in the femurs and vertebrae. Moreover, the introduction of SR 11302 under the experimental conditions is accompanied by an increase in the density and mineral saturation of the femurs and vertebrae, and an improvement in the biomechanical characteristics of the femurs (their strength and elasticity).

The Role of Different NO Synthase Isoforms in the Regulatory Effect of Prolactin and Growth Hormone on the State of Chromosomes in Mature Oocytes Aging in vitro

The fertility of female mammals depends on the quality of oocytes, which decreases in the process of biological aging. We found an inhibitory effect of two related pituitary hormones (prolactin (PRL) and growth hormone (GH)) on destructive modifications of metaphase chromosomes in the eggs of domestic cow (Bos taurus taurus), aging in vitro. The involvement of different NO synthase (NOS) isoforms in realization of the effect of PRL and GH on age-related changes of MII chromosomes with prolonged cultivation of in vitro matured cow oocytes was studied in the present work. In the absence of hormones NPLA, neuronal NOS (nNOS) inhibitor had no effect on the frequency of chromosomal abnormalities in aging oocytes. At the same time, 1400 W (inducible NOS (iNOS) inhibitor) and L-NAME (efficient endothelial NOS (eNOS and nNOS ) inhibitor) decreased this frequency, while the blocking of all three NO synthase isoforms led to the opposite effect. The inhibitory effect of PRL on destructive chromosome modifications in aging eggs increased in the presence of L-NAME, but was not expressed with the inhibition of nNOS, iNOS, or all NO synthase isoforms simultaneously. Neither NPLA nor L-NAME modulated the inhibitory effect of GH on abnormal chromosome changes in oocytes. On the contrary, the blocking of iNOS, as well as all three NO synthase isoforms, led to an increase in the portion of oocytes with destructive changes of MII chromosomes. At the same time, the level of total activity of NO synthase in oocytes did not depend on the presence of the studied hormones or NOS inhibitors during the prolonged cultivation of oocyte–cumulus complexes. Data obtained suggest that the inhibitory effect of PRL and GH on abnormal changes of metaphase chromosomes in aging cow eggs is associated with a decrease in the activity of endothelial NO synthase, as well as (in the case of PRL) with an increase in the activity of neuronal NO synthase in the cumulus cells surrounding the oocytes.

The response of nitric oxide system to high Altitude in Phrynocephalus erythrurus on Qinghai-Tibetan plateau.

Nitric oxide (NO), produced by nitric oxide synthases (NOS) from L-arginine, plays important roles in a wide range of physiological processes. However, little is known about ectothermic species. To investigate the response of NO/NOS system in adaptation to different altitudes in Phrynocephalus genus, the red tail toad-headed lizard Phrynocephalus erythrurus that live at 4500-5300 m on the Qinghai-Tibet Plateau and another low altitude living lizard Phrynocephalus przewalskii were selected in the present study. The results of mRNA expression and activity of NOSs, as well as NO metabolite levels in different tissues of the two lizards indicate that nNOS mRNA levels in cardiac and skeletal muscle were notably elevated in P. erythrurus, and iNOS expression was also increased markedly (up to 4-fold) in cardiac muscle. There was no significant difference in eNOS mRNA level in tested tissues between two species. However, the total NOS activity in skeletal muscle of P. erythrurus was slightly lower than that of P. przewalskii (p < .05) while no difference in other tissues. Similarly, lower iNOS activity (p < .01) was found in cardiac and skeletal muscle in P. erythrurus compared to P. przewalskii. In addition, the NO metabolite levels were dramatically lower in P. erythrurus in all tested tissues. We propose that higher nNOS and iNOS mRNA expression, lower iNOS activity and NO metabolite levels may represent physiological characteristics in nitric oxide system, which may contribute to high-altitude adaptation in P. erythrurus.