Plasma Nitric Oxide Concentration Research Articles

Hemp seed protein and its hydrolysate compared with casein protein consumption in adults with hypertension: a double-blind crossover study

BackgroundThe effects of consuming hemp seed protein (HSP) as well as its hydrolysate-derived bioactive peptide (HSP+) on blood pressure (BP) has not, to our knowledge, been investigated in humans. ObjectivesWe aimed to investigate how consumption of HSP and its hydrolysate modulates 24-h systolic (SBP) and diastolic BP (DBP) and plasma biomarkers of BP compared with casein. MethodsIn a double-blind, randomized, crossover design trial, 35 adults who had mild hypertension with SBP between 130 and 160 mmHg and DBP ≤110 mmHg were recruited. Participants were randomly assigned to varying sequences of 3 6-wk treatments, 50 g casein/d, 50 g HSP/d, or 45 g HSP plus 5 g HSP-derived bioactive peptides/d (HSP+), separated by a 2-wk washout period. Treatment effects were assessed with a linear mixed model with repeated measures. ResultsCompared with casein, after HSP+ consumption, 24-h SBP and 24-h DBP decreased from 135.1 and 80.0 mmHg to 128.1 ± 1.6 (P < 0.0001) and 76.0 ± 1.4 mmHg (P < 0.0001), respectively, whereas these values were 133.5 ± 1.6 and 78.9 ± 1.4 mmHg after HSP consumption (P < 0.0001). There were no differences between the HSP and HSP+ consumption in plasma angiotensin-converting enzyme (ACE) activity, renin, or nitric oxide (NO) concentrations. However, these 2 treatments were able to lower both ACE and renin activities and raise NO concentration in plasma compared with casein. ConclusionsThese results suggest that hemp protein consumption, as well as in combination with bioactive peptides, may have a role in the dietary management of hypertension.This trial was registered at clinicaltrials.gov as NCT03508895.

Effects of Phytosterol Ester Supplementation on Egg Characteristics, Eggshell Ultrastructure, Antioxidant Capacity, Liver Function and Hepatic Metabolites of Laying Hens during Peak Laying Period.

The aim of this experiment was to investigate the effects of dietary Phytosterol Ester (PSE) supplementation on egg characteristics, eggshell ultrastructure, antioxidant capacity, liver function, hepatic metabolites, and its mechanism of action in Hy-Line Brown laying hens during peak laying period. A total of 256 healthy Hy-Line Brown laying hens were randomly allocated into four groups. The hens in the control group were fed a basal diet, while those in the experimental groups were fed a basal diet further supplemented with 10, 20, and 40 mg/kg PSE, respectively. It was found that the addition of 20 mg/kg and 40 mg/kg PSE to the diets increased egg weight, but decreased egg breaking strength (p < 0.05). The addition of PSEs to the diets increased albumen height and Haugh unit in all experimental groups (p < 0.05). Electron microscopic observation revealed that the mammillary thickness increased significantly at doses of 20 and 40 mg/kg, but the total thickness decreased, and the effective thickness also thinned (p < 0.05). The mammillary width narrowed in all experimental groups (p < 0.001). Dietary supplementation with 40 mg/kg PSE significantly increased egg yolk Phenylalanine, Leucine, and Isoleucine levels (p < 0.05). In untargeted liver metabolomic analyses, L-Phenylalanine increased significantly in all experimental groups. Leucyl-Lysine, Glutamyl-Leucyl-Arginine, and L-Tryptophan increased significantly at doses of 10 and 20 mg/kg (p < 0.05), and L-Tyrosine increased significantly at doses of 10 and 40 mg/kg (p = 0.033). Aspartyl-Isoleucine also increased significantly at a dose of 10 mg/kg (p = 0.044). The concentration of total protein in the liver was significantly higher at doses of 20 and 40 mg/kg than that of the control group, and the concentrations of total cholesterol and low-density lipoprotein cholesterol were significantly reduced (p < 0.05). The concentration of triglyceride and alkaline phosphatase were significantly reduced in all experimental groups (p < 0.05). Steatosis and hemorrhage in the liver were also improved by observing the H&E-stained sections of the liver. Concerning the antioxidant capacity in the liver, malondialdehyde concentration was significantly reduced (p < 0.05) at a dose of 40 mg/kg. In the ovary, malondialdehyde and nitric oxide concentrations were significantly reduced (p < 0.001). In all the experimental groups, plasma nitric oxide concentration was significantly decreased while superoxide dismutase was significantly increased, and total antioxidant capacity concentration was significantly increased (p < 0.05) in the 10 mg/kg and 40 mg/kg doses. Metabolomics analyses revealed that PSEs play a role in promoting protein synthesis by promoting Aminoacyl-tRNA biosynthesis and amino acid metabolism, among other pathways. This study showed that the dietary addition of PSEs improved egg characteristics, antioxidant capacity, liver function, and symptoms of fatty liver hemorrhagic syndrome in Hy-Line Brown laying hens at peak laying stage. The changes in liver metabolism suggest that the mechanism of action may be related to pathways such as Aminoacyl-tRNA biosynthesis and amino acid metabolism. In conclusion, the present study demonstrated that PSEs are safe and effective dietary additives as an alternative to antibiotics.

Betulin-rich hydroalcoholic extract of Daphne oleoides attenuates bleomycin-induced pulmonary fibrosis in rat

Background and objectivePulmonary fibrosis (PF) is a chronic and progressive respiratory disease representing the final stage of lung inflammatory disorders. Reactive oxygen species (ROS), an essential factor in the formation and progression of pulmonary fibrosis, are a significant adverse effect of Bleomycin (BLM). Antioxidant activities have been found in Daphne oleoides. In this study, we attempted to explore the function of hydroalcoholic extract of Daphne oleoides (D. oleoides) and Betulin in inhibiting bleomycin (BLM)-induced pulmonary fibrosis in rat". Materials and methodsThe current experimental study used 36 male Wistar rats (180–220). Following a random process, the animals were divided into six groups six (n = 6). Group, I (the control group) received normal saline, while Group II (the hazardous group) received intratracheal BLM (7.5 units per kg). Following the administration of BLM, Groups V and VI received daily doses of vitamin E (500 mg/kg/d, p.o.) and Betulin (10 mg kg/d, p.o.), whereas Groups III and IV received daily doses of Daphne oleoides extract (300 and 600 mg/kg/d, p.o.). Then, blood samples from the hearts of the animals were taken to assess the plasma concentrations of nitric oxide (NO) and malondialdehyde (MDA). Finally, the rats were euthanized, and the lung tissues were taken out for histological analysis and assessments of the levels of lung hydroxyproline (HP), ferric-reducing ability (FRAP), NO, Glutathione Concentration (GSH), thiol content (tSH) and MDA. FindingsElevated lung index, lung hydroxyproline, NO, and MDA plasma levels, and a reduction in total body thiol content (tSH) in the group receiving BLM were evidence of pulmonary toxicity. Treatment with D. oleoides extracts, Betulin, and Vit E, especially at 600 mg/kg, led to a marked reduction in the above parameters compared with the BLM-received group (p < 0.01). Histological Analysis of the BLM-treated group showed a considerable Lung injury with interstitial infiltration, collapsed alveolar spaces, and alveolar septal thickening. These changes were mitigated with D. oleoides 600, Betulin-, and vitamin E. These changes were mitigated with D. oleoides 600, Betulin-, and vitamin E. ConclusionThese findings suggest that D. oleoides and Betulin prevent bleomycin-induced lung fibrosis in rats by decreasing inflammatory and antioxidant markers. Daphne oleoides, therefore, have the potential to be used therapeutically to treat pulmonary fibrosis.

The effect of exenatide (a GLP-1 analogue) and sitagliptin (a DPP-4 inhibitor) on asymmetric dimethylarginine (ADMA) metabolism and selected biomarkers of cardiac fibrosis in rats with fructose-induced metabolic syndrome

Asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide (NO) synthesis, is a risk factor for endothelial dysfunction, a common pathophysiological denominator for both atherogenesis and cardiac fibrosis.We aimed to investigate whether the cardioprotective and antifibrotic effects of incretin drugs, exenatide and sitagliptin, may be associated with their ability to affect circulating and cardiac ADMA metabolism.Normal and fructose-fed rats were treated with sitagliptin (5.0/10 mg/kg) or exenatide (5/10 µg/kg) for 4 weeks. The following methods were used: LC-MS/MS, ELISA, Real-Time-PCR, colorimetry, IHC and H&E staining, PCA and OPLS-DA projections.Eight-week fructose feeding resulted in an increase in plasma ADMA and a decrease in NO concentration. Exenatide administration into fructose-fed rats reduced the plasma ADMA level and increased NO level. In the heart of these animals exenatide administration increased NO and PRMT1 level, reduced TGF-ß1, α-SMA levels and COL1A1 expression. In the exenatide treated rats renal DDAH activity positively correlated with plasma NO level and negatively with plasma ADMA level and cardiac α-SMA concentration. Sitagliptin treatment of fructose-fed rats increased plasma NO concentration, reduced circulating SDMA level, increased renal DDAH activity and reduced myocardial DDAH activity. Both drugs attenuated the myocardial immunoexpression of Smad2/3/P and perivascular fibrosis.In the metabolic syndrome condition both sitagliptin and exenatide positively modulated cardiac fibrotic remodeling and circulating level of endogenous NOS inhibitors but had no effects on ADMA levels in the myocardium.

Dietary ARA, DHA, and Carbohydrate Ratios Affect the Immune Status of Gilthead Sea Bream Juveniles upon Bacterial Challenge.

This study aims to assess the effects of different dietary n-6/n-3 long-chain polyunsaturated fatty acid ratios and CHO content in the immune response of gilthead seabream. For that purpose, gilthead sea bream juveniles (initial body weight = 47.5 g) were fed for 84 days with four isoproteic (47% crude protein) and isolipidic (18% crude lipids) diets with high (20%) or low (5%) level of gelatinized starch (HS or LS diets, respectively) and included approximately 2.4% ARA or DHA. At the end of the trial, the DHA-enriched groups presented increased red blood cell (RBC) count, hemoglobin, plasmatic nitric oxide (NO) content, and antiprotease and alternative complement activities. The ARA groups had increased thrombocyte count, and plasmatic bactericidal activity against Vibrio anguillarum was lower in the fish fed the ARA/LS diet. After the feeding trial, the fish were challenged with an intraperitoneal injection (i.p.) of killed Photobacterium damselae subsp. piscicida (Phdp) and sampled at 4 and 24 h after the challenge. At 4 h after i.p., the ARA groups presented increased plasma total immunoglobulins (Ig) and bactericidal activity against V. anguillarum. In addition, the fish fed the ARA/LS diet presented lower white blood cell (WBC) and alternative complement activity. At 24 h after i.p., the ARA groups presented increased RBC, WBC, and thrombocyte numbers, total IG, plasma peroxidase activity, and casp3 expression in the distal intestine. The HS groups presented increased plasma NO content and bactericidal activity against Phdp and decreased protease, antiprotease activity, and bactericidal activity against V. anguillarum. In conclusion, high dietary DHA levels seemed to improve the immune status of unchallenged gilthead sea bream juveniles, while high dietary ARA levels improved the fish immune response to a bacterial challenge. The energy provided by dietary starch seems to be important to promote a fast response by the fish immune system after a challenge.

Effects of the In ovo Administration of L-Ascorbic Acid on Tissue L-Ascorbic Acid Concentrations, Systemic Inflammation, and Tracheal Histomorphology of Ross 708 Broilers Subjected to Elevated Levels of Atmospheric Ammonia

The effects of in ovo injection of L-ascorbic acid (L-AA) on tissue L-AA concentrations, systemic inflammation, plasma mineral concentrations, and tracheal histomorphology of Ross 708 broilers subjected to elevated atmospheric ammonia (NH3) levels after hatch were investigated. The four in ovo treatments included non-injected (control), saline-injected (control), or saline containing 12 or 25 mg of L-AA. The in ovo treatments were applied at 17 days of incubation by injecting a 100 μL volume of each pre-specified treatment into the amnion. At hatch, 12 male chicks were randomly allocated to each of the 12 replicate battery cages belonging to each treatment group. The cages were arranged in a randomized complete block design within a common room. All birds were exposed to 50 ppm of NH3 at 35 days of posthatch age (doa), and the concentration of NH3 in the room was recorded every 20 s. At 0, 7, 14, 21, and 28 doa, one bird from each cage was arbitrarily selected and euthanized for determinations of liver and eye L-AA concentrations at 0, 7, 14, 21, 28 doa; plasma nitric oxide concentrations at 0, 14, 21, and 28 doa; and plasma calcium and trace mineral concentrations at 0 and 21 doa. Tracheal histomorphology evaluations were performed at 0, 21, and 28 doa. There were no significant treatment differences for plasma nitric oxide and mineral concentrations, and for liver and eye L-AA concentrations at each sampling timepoint. In ovo injection of either 12 or 25 mg of L-AA decreased tracheal attenuation incidence at 0 doa compared to the non-injected or saline-injected control groups. Furthermore, the percentage of mild tracheal inflammation scores was lower at 28 doa in response to the in ovo injection of 12 mg of L-AA compared to the non-injected or saline-injected control groups. These results indicate that in ovo injection of 12 mg of L-AA reduces tracheal inflammation in broilers subjected to elevated atmospheric NH3.

Effects of dietary camelina, flaxseed, and canola oil supplementation on transepidermal water loss, skin and coat health parameters, and plasma prostaglandin E2, glycosaminoglycan, and nitric oxide concentrations in healthy adult horses

Abstract Camelina oil is derived from a low-input, high-yield crop and, in comparison to many other dietary fat sources currently used in equine diets, provides a greater amount of α-linolenic acid [ALA; (n-3)], than linoleic acid [LA; (n-6)]. However, no research exists assessing the effects of feeding camelina oil to horses in contrast to other commonly used oils. The objective of this study was to compare the effect of supplementing camelina oil to that of flaxseed and canola oil supplementation, on outcomes related to skin and coat health in horses. Thirty adult horses [23 mares, 7 geldings; 14.9 years ± 5.3 years; 544 ± 66 kg body weight (BW) (mean ± SD)] underwent a 4-week wash-in period consuming hay and sunflower oil. Following the wash-in period, horses were blocked by location, age, and BW, and assigned to one of three treatment oils for 16 weeks (370 mg oil/kg BW): camelina (CAM), canola (OLA), or flaxseed (FLX) oil. Blood samples were collected and plasma prostaglandin E2 (PGE2; ELISA), nitric oxide (NO; Griess Reaction), and glycosaminoglycan (GAG; DMMB) concentrations were measured on weeks 0 (n = 30), 14 (n = 24), and 16 (n = 30). On weeks 0, 2, 4, 8, and 16, transepidermal water loss (TEWL) was measured pre- and post-acetone application using a VapoMeter (n = 26), and a 5-point-Likert scale was used to assess skin and coat characteristics on the side and rump of the horses (n = 30). All data were analyzed with repeated measures ANOVA using PROC GLIMMIX in SAS. Independent of treatment, coat color, and quality increased from baseline. There were no differences in the outcomes assessed between the horses supplemented camelina oil and those supplemented canola or flaxseed oil. These results suggest that independent of treatment, all oil supplements improved coat color and quality in horses. This provides indication that camelina oil is comparable to existing plant-based oil supplements in supporting skin and coat health and inflammation in horses.

Effects of arginine and guanidinoacetic acid with or without phenylalanine on ascites susceptibility in cold-stressed broilers fed canola meal-based diet

In order to evaluate the effects of ARG sources (arginine [ARG] and Guanidinoacetic acid [GAA]) and phenylalanine (PHE) supplementation on performance, susceptibility to ascites, intestinal morphology, and nutrient digestibility in the cold-stressed broilers fed a canola meal (CM)-based diet, a 2×2 factorial experiment with four treatments was conducted. The dietary treatments included CM-based diet + 2.57 g/kg ARG, CM-based diet + 2.57 g/kg ARG + 1.5 g/kg PHE, CM-based diet + 1.8 g/kg GAA and CM-based diet + 1.8 g/kg GAA + 1.5 g/kg PHE. The corn-CM diet without supplementation was used as a negative control (NC) group in the fifth treatment that excluded the factorial arrangement. The results showed that adding ARG to diets without PHE supplement increased (p < 0.05) feed intake. Also, birds fed diets containing ARG had higher (p < 0.05) body weight gain (BWG) compared to those fed GAA added diets. Supplementation of PHE improved (p < 0.05) the FCR compared to groups fed diets without added PHE. Further, ARG addition increased (p < 0.05) plasma nitric oxide (NO) concentration, carcass, breast and leg yields, duodenal, jejunal, and ileal villus height (VH) to crypt depth (CD, and dry matter digestibility, while decreasing (p < 0.05) ascites mortality and right ventricle (RV) to total ventricle (TV) ratio compared to GAA added groups. Supplementation of PHE also declined susceptibility to ascites by reducing (p < 0.01) RV to TV ratio while increasing (p < 0.05) plasma NO level. The digestibility of ether extract also increased (p < 0.05) in broilers fed GAA supplemented diets versus those fed ARG added diets. The findings suggested that ARG may improve BWG and lower ascites incidence in broilers fed a diet based on CM under cold stress because of its antihypertensive effects. Moreover, the findings of this study demonstrated the importance of including PHE formulation in ARG-deficient diets to attenuate the adverse effects of cold stress on broilers. It was also concluded that GAA could be efficaciously used in cold-stressed broilers fed an ARG-deficient diet.

Evaluation of erythrocyte arginase activity, plasma nitric oxide concentration and oxidative stress status in cattle with anaplasmosis

Bovine anaplasmosis is an arthropod-borne disease characterized by high fever, anaemia and sometimes jaundice. The role of oxidative stress in anaplasmosis has been investigated, but erythrocyte arginase (ARG) activity has not been studied. In this study, we aimed to investigate the changes in haematological parameters, erythrocyte ARG activity, plasma nitric oxide (NO) levels and oxidative stress parameters and explain the relationship between each other in cattle with anaplasmosis. The material of this study consisted of 14 cattle, aged 10–12 months with anaplasmosis (infected group) and 14 healthy cattle aged 10–12 months (control group). Our data revealed that leukocyte parameters and plasma NO levels and serum malondialdehyde (MDA), total oxidant status (TOS) and oxidative stress index (OSI) levels were higher while erythrocyte parameters, erythrocyte ARG activity and serum total antioxidant status (TAS) and glutathione (GSH) levels were lower in the infected group compared to the control group. There was a strong correlation between erythrocyte ARG activity and NO, MDA, TOS, OSI, TAS and GSH. ROC analysis and correlation results suggest that erythrocyte ARG activity is an effective oxidative stress marker. We concluded that severe oxidative stress occurs in anaplasmosis. As the severity of anaemia increases, erythrocyte ARG activity plummets while plasma NO level elevates. These two parameters may also be used as prognostic and oxidative stress markers. Although decreased erythrocyte ARG activity is a disadvantage in haemolytic diseases, this situation can be compensated by increased NO. Thus, homeostasis of these two parameters may contribute to the elimination of the infection.

Intramuscular administration of l-arginine boosts testicular hemodynamics, plasma concentrations of testosterone and nitric oxide in heat-stressed rams

The current study aimed to assess, for the first time, the effects of intramuscular injection of l-arginine (L-arg) on testicular hemodynamics, echogenicity, and plasma concentrations of testosterone, total antioxidant capacity, and nitric oxide (NO) in Ossimi rams. Twelve sexually matured heat-stressed rams were randomly assigned to one of two groups: the L-arg group (n = 6) received 5 mg/kg L-arg dissolved in 2 ml normal saline 0.9%, or the control group (n = 6) received merely 2 ml of normal saline 0.9%. Blood sampling, B-mode ultrasound assessment of the testicular parenchyma, and pulsed-wave Doppler ultrasound of the testicular artery for both right and left testis were performed immediately before 0 min and 1, 4, 24, 48, 72, and 168 h after L-arg or saline administration. In the L-arg group, resistive index (RI) and pulsatility index (PI) means were significantly lower compared to the control group at 4–168 h post-treatment. Plasma testosterone concentrations were higher (P < 0.05) at 4 h and onward in the L-arg treated compared to the control rams, the same for NO levels however its increase (P < 0.05) was observed as soon as 1 h post-treatment. In L-arg treated rams, NO concentrations were positively correlated to plasma testosterone concentrations (r = 0.7, p < .01), but negatively correlated to both RI and PI (r = −0.8 and −0.6, respectively, p < .01). In conclusion, l-arginine administration enhanced testicular blood flow and increased plasma testosterone and nitric oxide concentrations in heat-stressed rams.

Dietary Strategies to Modulate the Health Condition and Immune Responses in Gilthead Seabream (Sparus aurata) Juveniles Following Intestinal Inflammation.

Several feed additives have proved to be beneficial in eliciting fish health. Β-glucans and curcumin are compounds with immunomodulatory capacities known to increase growth performance, stimulate immunity, improve general health, and enhance disease resistance in fish. The present study aimed to evaluate the effects of dietary Phaeodactylum tricornutum extracts rich in β-glucans and curcumin on gilthead seabream health status prior to and following an intestinal inflammatory stimulus. Three experimental diets were formulated: a practical commercial-type diet (CTRL), a CTRL diet supplemented with 1% microalgae-derived β-glucans extract (BG), and a CTRL diet supplemented with 0.2% of curcumin (CUR). After 30 days of the feeding trial, fish were sampled and subjected to an oral administration of 1% dextran sodium sulphate (DSS) to induce intestinal inflammation. Four groups were considered: a group of fish continued to be fed on the CTRL diet while the remaining groups were exposed to DSS, including CTRL-D (CTRL + DSS), BG-D (BG + DSS), and CUR-D (CUR + DSS), for 6 days. Growth, plasma and gut humoral immunity, liver and gut oxidative stress biomarkers, and intestinal gene expression were evaluated. No significant differences were found in growth after 30 days of feeding; however, seabream fed BG had decreased anti-protease activity and nitric oxide concentration in plasma while those fed CUR had increased mRNA levels of the tnfα, csf1r, and hep genes compared to those fed CTRL. After the inflammatory stimulus, hematocrit was enhanced in fish fed BG-D and CUR-D while red blood cell counts increased in those fed CTRL-D. Superoxide dismutase activity decreased in the intestine of all DSS groups while lipid peroxidation increased in the gut of fish fed CTRL-D and BG-D compared to CTRL. Moreover, the mRNA expression levels of csfr1 and sod decreased in fish fed CTRL-D and BG-D compared to CTRL, respectively. Despite the mild intestinal inflammatory condition induced by DSS, CUR was able to partially ameliorate its effects, improving the hematological profile and assisting against the oxidative stress.

Algorithm for predicting sepsis in newborns with respiratory pathology and perinatal lesions of the central nervous system on mechanical ventilation

BACKGROUND: Predicting sepsis in ventilated neonates remains a challenge in neonatology.&#x0D; AIM: To increase the efficiency of predicting sepsis diagnosis in newborns by developing a decision rule for its development based on decision trees.&#x0D; MATERIALS AND METHODS: This clinical study retrospectively reviewed 200 full-term newborns with respiratory pathology that are admitted to the intensive care unit and are on mechanical ventilation without clinical signs of bacterial infection.&#x0D; Upon admission to the department on days 1, 35, and 20, an enzyme-linked immunosorbent assay determined the plasma concentration of interleukin (IL)-1, IL-6, IL-8, tumor necrosis factor-, granulocyte colony-stimulating factor, soluble Fas ligand, fibroblast growth factors, and nitric oxide (NO), and immunophenotyping method determined CD3+CD19, CD3CD19+, CD3+CD4+, CD3+CD8+, CD69+, CD71+, CD95+, HLADR+, CD34+, CD14+, CD3CD56+; lymphocytes expressing AnnexinV-FITC+PI, and AnnexinV-FITC+PI+. The possibility of diagnosing sepsis upon intensive care unit admission was assessed by statistical cluster analysis of the total studied immunological criteria. The method of decision trees in the statistical environment R formed a diagnostic rule for predicting sepsis.&#x0D; RESULTS: Visualization of the cluster analysis results of admitted patients did not exclude the presence of two clusters among them (with and without sepsis, which explain the 60.81% of the point variability).&#x0D; Sepsis prediction rule are as follows: disease progression occurs if on day 1 CD95 is 16.8% and NO is 9.6 mkmol/l or CD95 is 16.8%, CD34 is 0.2%, CD69 is 4.12% or CD95 is 16.8%, CD34 is 0.2%, CD69 is 4.12%, and lymphocytes expressing AnnexinV-FITC+PI is 12.3%. The diagnostic accuracy was 96.00%; sensitivity was 97.00%; specificity was 94.90%; the false-positive proportion of diagnoses was 5.10%; the false-negative proportion of diagnoses was 2.94%; the positive result accuracy was 95.19%; and the negative result was 96.88%. The disease was complicated by bacterial sepsis development on 45 days of observation in 45 newborns.&#x0D; CONCLUSIONS: Significant importance in sepsis development belongs to the prevalence of altered immunocompetent cells over proliferation and endogenous synthesis of nitric oxide. The cumulative determination of CD95+, CD69+, AnnexinV-FITC+PI, CD34+, and plasma nitric oxide concentration helped diagnose sepsis development at the preclinical stage. The obtained results indirectly confirm the relevance of studies on sepsis prevention and treatment by drug correction of apoptosis and inhaled NO.

0153 Bright light exposition potentiates the vasodilation promoted by dynamic handgrip exercise

Abstract Introduction Bright light (~5000 lux) directed at human skin increases plasma nitric oxide concentration promoting vasodilation, with increased skeletal muscle blood flow (BF) and decreased blood pressure (BP). The same stimulus directed at the eyes increases sympathetic activity, promoting vasoconstriction and increased BP. If bright light potentiates or mitigates the vasodilation promoted during exercise is unknown. Therefore, this study aimed to compare the effect of different intensities of lights on vasodilation of the active skeletal muscle during dynamic handgrip exercise. Methods Eleven healthy and physically inactive adult men (29±6 years) participated of the current study. All experiments were conducted between June and July 2021, beginning at 5 PM and under constant environmental conditions. Subjects performed dynamic handgrip exercise using the dominant arm at a 90º angle in the supine position with 2 s cycles of contraction and relaxation for 6 min at 40% of maximal voluntary contraction during the three experimental condition in a randomized order: (BL) 5000 lux, Control light (CL) 500 lux, and Dim light (DL) ≤8 lux. In each condition, (BL, CL or DL), subjects remained in a rested state for 20 min before 1 min of baseline assessment followed by exercise. Conditions were always separated by 20 min of washout (under CL). Assessments comprised BF, vascular conductance (VC), and diameter of brachial artery of exercising arm (Ultrasound); BP measured in the rest arm (Photoplethysmography) and heart rate (ECG). Two-way ANOVA for repeated measures, p≤0.05. Results Baseline were not different (p&amp;gt;0.05). During exercise, BF and VC increased in the three conditions, however it was potentiated by BL (p &amp;lt; 0.0001 and p &amp;lt; 0.0001, respectively) compared with CL (+19% and +15%, respectively) and with DL (+12% and +11%, respectively). Arterial diameter increased similarly in the three conditions. Mean BP increased in the three conditions, however it was attenuated in the BL compared with DL (-23%) and similar to CL (&amp;lt;0.0001). Conclusion Bright light can potentiate skeletal muscle vasodilation during a small-mass muscle exercise attenuating the increase of BP. Support (If Any) The São Paulo Research Foundation - FAPESP 2020/11588-2.

L-arginine supplementation improved neonatal outcomes in pregnancies with hypertensive disorder or intrauterine growth restriction: A systematic review and meta-analysis of randomized controlled trials

Previous research established that the availability of l-arginine affects placental vascular development and fetal growth. However, practical details associated with the effects of l-arginine supplementation on the neonatal outcomes of hypertensive disorder (HD) and intrauterine growth restriction (IUGR) pregnancies are limited. The PubMed, ScienceDirect, and Web of Science databases were searched for peer-reviewed literature published by September 30, 2021 to investigate the operational details of l-arginine supplementation in improving neonatal outcomes in complicated pregnancies. Standardized mean difference (SMD) and weighted mean difference (WMD) of continuous variables, as well as the risk ratio (RR) for categorical variables were pooled by random-effects models. The results indicated that l-arginine supplementation increased the plasma nitric oxide (NO) concentrations in IUGR pregnancies (SMD: 0.71; 95% CI: 0.45, 0.97; I2=0%), but decreased the risk of preeclampsia in HD mothers (RR: 0.49; 95% CI: 0.31, 0.76; I2=0%). Administration with l-arginine elevated birth weights both in hypertensive and IUGR pregnant women, with WMDs of 194.70g (95% CI: 58.21, 331.20; I2=44.2%) and 134.00g (95% CI: 43.53, 224.46; I2=42.4%), respectively. However, the intervention had no effect on gestational age except in HD pregnancies (WMD: 7.05d; 95% CI: 3.16, 10.95; I2=36.5%). l-arginine administration during pregnancy significantly reduced the small for gestational age (SGA) risk of fetus both in HD (RR: 0.51; 95% CI: 0.31, 0.83; I2=0.0%) and IUGR mothers (RR: 0.46; 95% CI: 0.25, 0.88; I2=0.0%). Subgroup analyses revealed that l-arginine supplementation at <4g/d dosage or for ≥1-month duration or in the third trimester had a greater effect on birth weights in HD women without proteinuria, but a higher l-arginine dosage was more beneficial for extending gestational age and reducing the risk of SGA in older pregnancies. Additionally, intravenous infusion of l-arginine, but not oral administration, significantly increased birth weight in IUGR pregnancies with elevated NO concentrations, although the recommended amount should be confined to <4g/d. These findings provide practical guidelines for l-arginine supplementation to improve the birth outcomes of complicated pregnancies. CRD42021246290 (https://www.crd.york.ac.uk/PROSPERO).

ENOS T786C (rs2070744) Gene Polymorphism and Its Underlying Mechanism Associated with Coronary Heart Disease

The study aims to examine plasma nitric oxide concentration and its relationship to T-786C gene polymorphism levels in the development of coronary heart disease in young people. Reduced nitric oxide (NO) bioavailability and endothelial nitric oxide synthase (eNOS) T786C gene polymorphism have been identified as risk factors for the development of coronary heart disease. This cross-sectional study was conducted in SRM Medical College Hospital and Research Centre on 200 angiographically proven CHD subjects attending the Department of Cardiology and medicine and 100 controls from MHC in the age group of ≤ 45 years. Overnight fasting plasma samples were obtained to evaluate lipid profile and nitric oxide by Griess reaction in UV spectrophotometry utilizing the ELISA technique. Polymerase Chain Reaction and Restricted fragment length polymerase amplify the eNOS gene T-786C. As a result, NO levels in plasma were significantly lower in CHD patients than in controls. In addition, C allele carriers of the eNOS T786C polymorphism showed significantly lower mean plasma NO levels than T allele carriers (P &lt; 0.001). Our findings suggest that a lower plasma NO level is related to an increased risk of CHD. Furthermore, the eNOS T786C polymorphism is a significant risk factor for CHD development by lowering NO levels in the plasma. However, the eNOS T786C polymorphism influences the severity of CHD.

Effects of the In Ovo Injection of Vitamin D3 and 25-Hydroxyvitamin D3 in Ross 708 Broilers Subsequently Challenged with Coccidiosis: II Immunological and Inflammatory Responses and Small Intestine Histomorphology.

Simple SummaryVitamin D3 sources serve as immunomodulators and improve intestinal morphology, which can promote broiler performance. The in ovo injection of vitamin D3 sources has been shown to enhance immunity as well as histomorphological variables in unchallenged conditions. One of the main diseases affecting poultry production is coccidiosis. Because of this, the current study was designed to determine the effects vitamin D3 (D3) and 25-hydroxyvitamin D3 (25OHD3) alone or together on the inflammatory reaction and small intestine morphology of broilers that were challenged with coccidiosis. In this study, it is shown that the in ovo administration of 2.4 μg of 25OHD3 alone increased villus length to crypt depth ratio (VCR) with the D3 + 25OHD3 treatment being intermediate two weeks post-challenge (28 day of age). Furthermore, chickens that received of 25OHD3 alone experienced lower plasma nitric oxide concentration as a systemic inflammatory indicator in comparison to all other treatments. It is concluded that the in ovo injection of 2.4 μg of 25OHD3 at 18 days of incubation can enhance intestinal histomorphology as well as inflammatory reaction of broilers when infected with coccidiosis.In broilers challenged with coccidiosis, effects of in ovo vitamin D3 (D3) and 25-hydroxyvitamin D3 (25OHD3) administration on their inflammatory response and small intestine morphology were evaluated. At 18 d of incubation (doi), a 50 μL volume of the following 5 in ovo injection treatments was administrated: non-injected (1) and diluent injected (2) controls, or diluent injection containing 2.4 μg D3 (3) or 2.4 μg 25OHD3 (4), or their combination (5). Four male broilers were randomly allocated to each of eight isolated replicate wire-floored battery cages at hatch, and birds were challenged at 14 d of age (doa) with a 20x live coccidial vaccine dosage. One bird from each treatment–replicate (40 birds in each of 8 replicates per treatment) was bled at 14 and 28 doa in order to collect blood for the determination of plasma IL-1β and nitric oxide (NO) concentrations. The duodenum, jejunum, and ilium from those same birds were excised for measurement of villus length, crypt depth, villus length to crypt depth ratio (VCR), and villus surface area. In ovo injection of 2.4 μg of 25OHD3 resulted in a reduction in plasma NO levels as compared to all other treatments at 28 doa. Additionally, duodenal VCR increased in response to the in ovo injection of 25OHD3 when compared to the diluent, D3 alone, and the D3 + 25OHD3 combination treatments at two weeks post-challenge (28 doa). Therefore, it can be concluded that 2.4 μg of 25OHD3, when administrated in ovo at 18 doi, may be used to decrease the inflammatory reaction as well as to enhance the small intestine morphology of broilers during a coccidiosis challenge.

Eight Days of L-Citrulline or L-Arginine Supplementation Did Not Improve 200-m and 100-m Swimming Time Trials.

The effects of L-citrulline or L-arginine supplementation on exercise performance are equivocal, and the effects on swimming performance are unclear. We aimed to assess whether 8-day supplementation with L-arginine or L-citrulline supplementation would improve 200 m and 100 m freestyle swimming time-trial performances. After the baseline trial (first visit), in a double-blind, randomised design, 15 trained/developmental (5 females) swimmers and triathletes were assigned to three groups and underwent an 8-day supplementation period, with a daily dose of either 8 gr L-arginine (Arg, n = 5) or L-citrulline (Cit, n = 5) or placebo (Pla, n = 5). On day 9, participants completed experimental trial (second visit). In each trial, after blood sampling, participants performed both 200 m and 100 m freestyle swimming time-trials, with 30 min recovery between trials. Plasma nitric oxide (NOx) and blood lactate concentrations (BLa) were collected immediately before and after 200 m and 100 m TTs, respectively. No significant difference was observed in NOx between groups (p = 0.201). There was no significant difference in 200 m (p = 0.226) and 100 m swimming time-trials (p = 0.993) between groups. There was a main effect of time on BLa concentration (p < 0.001), but no trial × group (p = 0.243) and trial × lactate × group interaction effect (p = 0.276) was present. Furthermore, 8-day either L-citrulline or L-arginine supplementation did not enhance middle (200 m) and short-distance (100 m) swimming performance in trained/developmental swimmers and triathletes. These findings do not support the use of L-citrulline or L-arginine supplementation as ergogenic aids for swimming performance.

Acetate-mediated-obestatin modulation attenuates adipose-hepatic dysmetabolism in high fat diet-induced obese rat model.

Approximately 650 million of world adult population is affected by obesity, which is characterized by adipose and hepatic metabolic dysfunction. Short chain fatty acids (SCFAs) have been linked to improved metabolic profile. However, the effect of SCFAs, particularly acetate on adipose-hepatic dysfunction is unclear. Therefore, the present study investigated the role of acetate on adipose-hepatic metabolic dysfunction and the possible involvement of obestatin in high fat diet-induced obese Wistar rats. Adult male Wistar rats (160-190 g) were allotted into groups (n = 6/group): Control, acetate-treated, obese and obese + acetate-treated groups received vehicle (distilled water), sodium acetate (200 mg/kg), 40% HFD and 40% HFD plus sodium acetate respectively. The administration lasted for 12 weeks. HFD caused increased body weight gain and visceral adiposity, insulin resistance, hyperinsulinemia and increased pancreatic-β cell function and plasma/hepatic triglyceride and total cholesterol as well as decreased adipose triglyceride and total cholesterol, increased plasma, adipose, and hepatic malondialdehyde, TNF-α, uric acid, lactate production and plasma/adipose but not gamma-glutamyl transferase and decreased plasma, adipose, and hepatic nitric oxide, glucose-6-phosphate dehydrogenase (G6PD), glutathione (GSH) and obestatin concentration compared to the control group. Notwithstanding, treatment with acetate attenuated the alterations. The results demonstrate that high fat diet-induced obesity is characterized with adipose and hepatic lipid dysmetabolism, which is associated with obestatin suppression. Findings also suggest that acetate provide protection against adipose and hepatic metabolic perturbations by restoring obestatin as well as G6PD/GSH-dependent antioxidant system.

Plasma and amniotic fluid concentrations of nitric oxide: Effects on uterine artery and placental vasculature in women who underwent voluntary pregnancy termination and in women with missed and threatened abortion. A pilot study

ObjectivesFirst trimester miscarriage is a multifactorial event. Various angiogenic factors have been proposed as possible early markers of non-viable pregnancies. The aim of the present study was to evaluate the systemic nitric oxide (NO) production in healthy early pregnancy and its possible role in first trimester miscarriage. Study designWe prospectively enrolled women referred to our Unit for elective termination of pregnancy, threatened abortion or missed abortion. Blood samples were taken for testing circulating NO plasma levels. Subsequently, all patients underwent 2-D ultrasonographic analysis and Color Doppler imaging to assess the pulsatility index of the uterine arteries. 3-D ultrasonographic and power Doppler analysis allowed a volumetric and vascular reconstruction of the placenta. During dilatation and vacuum aspiration, amniotic fluid was collected. ResultsSeventy-two patients were enrolled: 25 with elective termination of pregnancy (Group I); 17 with threatened abortion (Group II); 30 with missed abortion (Group III). Group II showed greater placental volume and lower uterine arteries PI than others. The plasma NO concentration resulted statistically higher in women with threatened abortion, while amniotic fluid NO concentration were higher in the viable pregnancies (Group I) than in the aborted fetuses (Group III). Plasma NO was inversely correlated with both mean arterial pressure and uterine artery PI and was positively correlated with amniotic fluid NO and CRL; amniotic fluid NO was positively correlated with placental Vascularization Index and Vascularization-Flow Index. ConclusionAmniotic NO concentration was higher in viable pregnancies and positively related to Doppler 3D indices of vascularization and blood flow within the placenta. Further studies are needed to elucidate its role in first trimester miscarriage.

Clinical evidence for elevated levels of caveolin-1 in circulation of patients with diabetic foot ulcers.

Caveolin-1 directly interacts vascular endothelial growth factor receptor-2 (VEGFR2) and therefore prevents VEGF-induced angiogenesis. In addition, the production of nitric oxide (NO), which is effective in reducing ischemia in diabetic foot ulcers (DFU), is suppressed by caveolin-1 in endothelial cells. The present study was designed to investigate the change of caveolin-1 concentrations in DFU patients. A total of 150 participants were consecutively enrolled, including 40 DFU patients (DFU group), 40 diabetes patients without DFU (type 2 diabetes mellitus [T2DM] group), and 70 participants without diabetes (control group). Significant increased levels of plasma caveolin-1, accompanied with decreased concentration of plasma VEGF-A (vascular endothelial growth factor-A) and NO, were detected in DFU patients. Moreover, Pearson's correlation analysis revealed a negative correlation between plasma caveolin-1 and VEGF-A as well as NO levels in DFU patients. Furthermore, DFU patients had higher expression of caveolin-1 in the popliteal artery, compared to those in control and T2DM groups. Simultaneously, the amounts of eNOS (an enzyme responsible for the production of NO) and VEGFR2 were attenuated in the popliteal artery of DFU patients. Taken together, our study provided clinical evidence for the possible association of elevated caveolin-1 levels and the development of DFU. This may be induced by the suppressed VEGF-A/VEGFR2 and eNOS/NO signalling axis.