Proteolytic enzyme-treated seaweed co-product (Ulva laetevirens) inclusion in corn-soybean and European broiler diets to improve digestibility, health, and performance.

The impact of addition of an Ulva laetevirens (previously Ulva rigida) co-product treated with a broad-spectrum endo-protease when added to a standard corn-soy (S) based diet and a diet based on European protein sources (EU) on performance, in vivo digestibility and multiple gastrointestinal characteristics was investigated in broilers. In total, 624 Ross 308 one-day-old male broilers were fed one of 6 experimental diets (8 replicates) consisting of a basal diet (S or EU), or a basal diet including the U. laetevirens co-product (U) treated without (U-) or with (U+) a proteolytic enzyme. Starter diets contained 0 (wk 1) and 2.5 (wk 2), and the grower diets (wk 3 and 4) 5% seaweed co-product. In the last 2 wk, birds fed the S vs. EU grower diets showed a higher BW, BWG, and FI, as well as a lower FCR (-0.05 g/g) in wk 3 (P < 0.05). Heavier gizzards (+13%; P < 0.001) and heavier gizzard contents (+92%; P < 0.001) were observed in birds fed the EU vs. S diets, as well as longer villi (+8%; P = 0.010). U diets had a higher water holding capacity than the basal diets (+19%). In wk 4, U inclusion resulted in increased FCR (+0.06 g/g; P < 0.001), water intake (+7%; P < 0.001), and duodenal cross section (+5%; P = 0.033). Enzyme treatment did not affect digestibility of any nutrients, except for ash which was increased in birds fed U+ vs. U- diets (+60%; P < 0.001). U in S diets led to higher, and U in EU diets led to lower apparent pre-cecal digestibility of all nutrients (P < 0.001 for all nutrients). Although for both diet types performance was decreased, dietary U. laetevirens inclusion had different effects when added to a standard corn-soy diet and a diet based on European protein sources. No obvious health effects were observed, leading to the conclusion of the absence of performance of health promoting bioactive components in the U. laetevirens co-product, or of diminishing of these effects due to the proteolytic enzyme treatment.

A proteolytic enzyme treatment to improve Ulva laetevirens and Solieria chordalis seaweed co-product digestibility, performance, and health in broilers.

To explore the potential use of seaweed co-products for broiler diets, this study investigates whether an enzyme treatment of seaweed co-products improves performance, in vivo digestibility and health in broilers. In total, 360 Ross 308 male broilers were fed one of 5 experimental diets: a basal diet, or a basal diet including the U. laetevirens or S. chordalis co-product, with or without proteolytic enzyme treatment of the seaweed, using 6 replicate pens of 12 birds each. The starter (d 0-13) and grower (d 14-21) diet contained 5 and 10% (w/w) seaweed product, respectively. A general linear model with contrast statements was used after model assumptions and goodness of fit were evaluated through normal distribution of residuals. Inclusion of seaweed in the broiler diets increased body weight gain (+14%; P = 0.002), and feed intake (+12%; P = 0.001) in the third week of the experiment. Birds fed the U. laetevirens compared to the S. chordalis diets had a higher body weight gain (+11%; P = 0.007), and a lower feed conversion ratio (FCR; -7%; P < 0.001). Seaweed inclusion reduced apparent pre-cecal digestibility of all nutrients (P < 0.05). Birds fed U. laetevirens vs. S. chordalis diets had a 10% reduced villus length (P < 0.001). Enzymatic treatment reduced the digestibility of most nutrients, and increased crypt depth in birds fed the U. laetevirens diets, whereas the opposite was observed for the birds fed the S. chordalis diets (Seaweed × Enzyme P = 0.035). Untreated vs. treated seaweed in the diets led to lower (-60%) plasma Interleukin-13 levels (P = 0.035). In conclusion, the proteolytic enzyme treatment of the seaweed co-products did not improve performance nor health-related parameters, and reduced digestibility of the diets. Dietary inclusion of U. laetevirens co-products did improve performance based on growth and FCR, whereas inclusion of S. chordalis did not. Inclusion of U. laetevirens in broiler diets slightly reduced duodenal villus length and crypt depth. The inflammation response was strongly reduced, specifically in birds fed the untreated U. laetevirens diet, making the U. laetevirens co-product of interest for future research.

Mortality After Elective and Ruptured Abdominal Aortic Aneurysm Surgical Repair: 12-Year Single-Center Experience of Estonia.

BACKGROUND AND AIMS: Abdominal aortic aneurysm is a degenerative vascular pathology with high mortality due to its rupture, which is why timely treatment is crucial. The current single-center retrospective study was undertaken to analyze short- and long-term all-cause mortality after operative treatment of abdominal aortic aneurysm and to examine the factors that influence outcome. MATERIAL AND METHODS: The data of all abdominal aortic aneurysm patients treated with open repair or endovascular aneurysm repair in 2004-2015 were retrospectively retrieved from the clinical database of Tartu University Hospital. The primary endpoint was 30-day, 90-day, and 5-year all-cause mortality. The secondary endpoint was determination of the risk factors for mortality. RESULTS AND CONCLUSION: Elective abdominal aortic aneurysm repair was performed on 228 patients (mean age 71.8 years), of whom 178 (78%) were treated with open repair and 50 (22%) with endovascular aneurysm repair. A total of 48 patients with ruptured abdominal aortic aneurysm were treated with open repair (mean age 73.8 years) at the Department of Vascular Surgery, Tartu University Hospital, Estonia. Mean follow-up period was 4.2 ± 3.3 years. In patients with elective abdominal aortic aneurysm, 30-day, 90-day, and 5-year all-cause mortality rates were 0.9%, 2.6%, and 32%, respectively. In multivariate analysis, the main predictors for 5-year mortality were preoperative creatinine value and age (p < 0.05). In patients with ruptured abdominal aortic aneurysm, 30-day, 90-day, and 5-year all-cause mortality rates were 22.9%, 33.3%, and 55.1%, respectively. In multivariate analysis, the risk factors for 30-day mortality in ruptured abdominal aortic aneurysm were perioperative hemoglobin and lactate levels (p < 0.05). According to this study, the all-cause mortality rates of elective abdominal aortic aneurysm and ruptured abdominal aortic aneurysm at our hospital were comparable to those at other centers worldwide. Even though some variables were identified as potential predictors of survival, the mortality rates after ruptured abdominal aortic aneurysm remain high. Early diagnosis, timely treatment, and detection of the risk factors for abdominal aortic aneurysm progression would improve survival in patients with abdominal aortic aneurysm.

Inverse relations of serum phosphatidylcholines and lysophosphatidylcholines with vascular damage and heart rate in patients with atherosclerosis.

BACKGROUND AND AIMS: The rapidly growing discipline of lipidomics allows the study of a wide spectrum of lipid species in body fluids and provides new insights into the pathogenesis of cardiovascular disease. We investigated serum phosphatidylcholine (PC) and lysophosphatidylcholine (lysoPC) species in relation to arterial stiffness, hemodynamics, and endothelial dysfunction in symptomatic patients with atherosclerosis and in healthy controls. METHODS AND RESULTS: Thirty-two patients with peripheral arterial disease (age 61.7 ± 9.0 years), 52 patients with coronary artery disease (age 63.2 ± 9.2 years), and 40 apparently healthy controls (age 60.3 ± 7.1 years) were studied. Serum levels of 90 glycerophospholipids were determined with the AbsoluteIDQ™ p180 kit (BIOCRATES Life Sciences AG, Innsbruck, Austria). The technique of applanation tonometry was used for non-invasive pulse wave analysis and carotid-femoral pulse wave velocity (cf-PWV) assessment. Decreased serum levels of several individual PC and lysoPC species (e.g., PC aa C28:1, PC aa C30:0, PC aa C32:2, PC ae C30:0 and PC ae C34:2, lysoPC a C18:2) were observed for the patient groups in comparison to the healthy subjects. In addition, a considerable number of PCs and lysoPCs were inversely related to either cf-PWV, heart rate, asymmetric dimethylarginine (ADMA) or ADMA/arginine for patients with symptomatic atherosclerosis but not for the controls. CONCLUSION: We found altered relationships between PC and lysoPC profiles, inflammation, and arterial function in atherosclerotic patients, compared to healthy subjects.

Association Between Fibulin-1 and Aortic Augmentation Index in Male Patients with Peripheral Arterial Disease.

BACKGROUND: Fibulin-1 (FBLN-1), a newly identified biomarker for vascular stiffness in type 2 diabetes, may participate in the pathophysiological processes leading to progression of arterial stiffness in atherosclerosis. In the present study, the relationship between FBLN-1 and arterial stiffness was examined in patients with atherosclerosis and in healthy subjects. METHODS: Thirty-eight patients with peripheral arterial disease (PAD) (age 62.4 ± 9.0 years), 38 patients with coronary artery disease (CAD) (age 64.0 ± 9.5 years), and 30 apparently healthy controls (age 61.1 ± 6.4 years) were studied. Serum FBLN-1, oxidized low density lipoprotein (oxLDL), resistin and plasminogen activator inhibitor-1 (PAI-1) levels were measured using the enzyme linked immunosorbent assay method. The technique of applanation tonometry was used for non-invasive pulse wave analysis and pulse wave velocity assessments. RESULTS: The levels of FBLN-1 (PAD = 9.4 [4.9-17.8] vs. CAD = 7.1 [4.8-11.8] vs. controls = 5.6 [4.1-8.4] µg/mL; p = .005), carotid-femoral pulse wave velocity (cf-PWV) (9.8 ± 2.2 vs. 9.5 ± 2.2 vs. 8.3 ± 2.2 m/s; p = .023) and the heart rate corrected augmentation index (AIx@75) (29.4 ± 7.2 vs. 19.2 ± 7.2 vs. 15.4 ± 7.1%; p < .001), differed among the three groups. A correlation between FBLN-1 and AIx@75 was observed only in patients with PAD (rho = 0.37, p = .021). The relationship retained statistical significance in a multiple regression model after adjustment for potential confounders. CONCLUSIONS: An independent association was demonstrated between serum FBLN-1 and AIx@75 in the PAD group. Thus, the findings suggest that FBLN-1 may play a role in arterial stiffening in patients with atherosclerosis.

In- and outdoor reproduction of first generation common sole Solea solea under a natural photothermal regime: Temporal progression of sexual maturation assessed by monitoring plasma steroids and gonadotropin mRNA expression.

Reproduction of many temperate fishes is seasonal and maturation and spawning of gametes are under photothermal control. Reproductive success of first generation (G1) common sole Solea solea in captivity has been low. In this study, the sexual maturation status has been assessed during the prespawning months in G1 sole that were housed (a) outdoor under the natural photoperiod and temperature, or (b) indoor under artificial photothermal induction. Maturation was assessed in male and female G1 broodstock in November as controls, after which the remaining population was divided over two outdoor flow-through tanks placed in a pond and two indoor recirculating aquaculture system (RAS) tanks. Subsequently, maturation status (gonadosomatic index GSI and plasma levels of testosterone T and 17ß-estradiol E2) was assessed in one tank for each condition in January, February and during spawning in early April, while fish in the other tank were not disturbed in achieving reproductive success. Quantitative real-time PCR was performed to determine species-specific gonadotropin mRNA expression in females. Successful G1 spawning and egg fertilisation occurred in all experimental tanks. Gonadal development was similar under both conditions. Higher E2 and T levels were found in indoor housed females. Gonadotropin expression revealed similar profiles between outdoor and indoor housed females. G1 sole could be reproduced in the outdoor tanks under the natural photoperiod and in the indoor tanks under artificial simulation of this regime that includes a potentially crucial chilling period of 2-3 months at 5-7 °C.

Prognostic impact of arterial stiffness in patients with symptomatic peripheral arterial disease.

OBJECTIVES: Arterial stiffness (AS) is increasingly recognized as an independent risk factor in different high-risk populations. Whether changes in AS can predict prognosis in patients with symptomatic peripheral arterial disease (PAD) has never been investigated. The aim of the present study was to test the hypothesis that AS is an independent predictor of all-cause and cardiovascular disease (CVD) mortality in patients with symptomatic PAD. METHODS: A cohort of 117 symptomatic PAD patients (aged 62.3 ± 7.7 years) were prospectively recruited from the Department of Vascular Surgery, Tartu University Hospital, between 2002 and 2010. The AS was measured using pulse wave analysis and assessment of pulse wave velocity (PWV). RESULTS: During the follow-up period (mean 4.1 ± 2.2 years) there were 32 fatal events. Kaplan-Meier analysis showed that the probability of all-cause and CVD mortality decreased with increasing small artery elasticity (SAE), as estimated by the log-rank test (p = .004; p = .005, respectively). By contrast, large artery elasticity, augmentation index, and aortic and brachial PWV were not significantly related to mortality. In a Cox proportional hazard model, SAE above the median was associated with decreased all-cause and CVD mortality after adjustment for confounding factors: relative risk (RR), 0.37; 95% confidence interval (CI), 0.17-0.81; p = .01; RR, 0.11; 95% CI, 0.01-0.86; p = .04, respectively). CONCLUSIONS: This study provides the first evidence, obtained from an observational study, that decreased small artery elasticity is an independent predictor of all-cause and CVD mortality in patients with symptomatic PAD.

Aortic stiffness and vitamin D are independent markers of aortic calcification in patients with peripheral arterial disease and in healthy subjects.

OBJECTIVE: Arterial stiffness is a significant determinant of cardiovascular risk and is related to vascular calcification. Vitamin D may regulate arterial calcification and has been associated with cardiovascular survival benefits. However, data about the relationship between arterial stiffness, aortic calcification and vitamin D levels in patients with peripheral arterial disease (PAD) and in healthy subjects are limited. We examined the potential association between aortic calcification, arterial stiffness and vitamin D levels in patients with symptomatic PAD and in healthy individuals. METHODS: We studied 78 men with PAD (aged 63 ± 7 years) and 74 healthy men (aged 61 ± 10 years). Aortic pulse wave velocity (aPWV) was determined by applanation tonometry using the Sphygmocor device. Aortic calcification score (ACS) was quantified by computed tomography. Serum 25-hydroxyvitamin D (25(OH)D) levels were measured using a radioimmune assay. RESULTS: ACS (4.9(2.3-8.9) vs. 0.2(0.03-1.6) (cm³); p < 0.01), aPWV (9.8 ± 2.4 vs. 8.2 ± 1.6 (m s⁻¹; p < 0.01) and 25(OH)D (15.1 ± 5.4 vs. 19.0 ± 5.9 (ng ml⁻¹); p < 0.01) were different in the patients compared with the controls. In multivariate analysis, ACS was independently determined by 25(OH)D, aPWV, calcium and age in patients with PAD (R² = 0.49; p < 0.001) and by 25(OH)D, aPWV, cholesterol/high-density lipoprotein (HDL) and age in the control group (R² = 0.55; p < 0.001). Increased aPWV and lower levels of 25(OH)D were associated with decreased ankle-brachial pressure index (p = 0.03). CONCLUSION: These results indicate that calcification of the aorta is independently associated with aortic stiffness and serum 25(OH)D level in patients with PAD and in healthy subjects. Aortic stiffness and abnormal vitamin D level may contribute to vascular calcification and are related to higher severity grade of atherosclerotic disease.

Effects of a synbiotic product on blood antioxidative activity in subjects colonized with Helicobacter pylori.

AIM: To evaluate the impact of the consumption of a synbiotic product on the antioxidative activity markers of blood in asymptomatic H. pylori-colonized persons. METHODS AND RESULTS: Fifty-three healthy adult volunteers without gastric symptoms participated in a randomized, double-blind placebo-controlled study. The crossover consumption of the enterocoated capsules containing antioxidative Lactobacillusfermentum ME-3, Lact. paracasei 8700:2 and Bifidobacterium longum 46 with Raftilose P95 lasted for 3 weeks and did not change the H. pylori colonization. In H. pylori-positive subjects the sera values of total antioxidative status (TAS) were significantly lower compared to H. pylori-negative subjects (0.97 vs 1.05 mmol l(-1), P = 0.008). After the consumption of the synbiotic, TAS values (0.97 vs 1.03 mmol l(-1), P = 0.004) increased, while the ratio between oxidized and reduced glutathione (0.035 vs 0.030, P = 0.016) decreased in H. pylori-positive subjects. CONCLUSIONS: The consumption of a synbiotic containing an antioxidative probiotic strain improved the reduced systemic antioxidative activity in H. pylori-colonized asymptomatic subjects. SIGNIFICANCE AND IMPACT OF THE STUDY: A synbiotic product containing an antioxidative probiotic strain may be useful in the reduction of systemic oxidative stress in H. pylori infection.

Ten years experience of treating aorto-femoral bypass graft infection with venous allografts.

OBJECTIVE: The aim of the present study was to analyze our results of treating synthetic vascular graft infection in the aorto-femoral segment with venous allografts. PATIENTS AND METHODS: During 1997-2006 we treated 11 male patients with venous allografts for aorto-femoral segment prosthetic graft infection. Patients were followed postoperatively with patency assessment based on Doppler ultrasonography. RESULTS: The mean interval between the implantation of the synthetic vascular graft and the appearance of infection was 43+/-29 months (range 5 months-10 years). Overall primary patency of the venous allograft was 100%, 88% and 66% at 1, 6 and 36 months, respectively. Secondary patency rates of the venous allograft were 94% and 73% after 5 and 8 years, respectively. Overall survival rate was 100%, 64%, 46% and 27% after one month, 3, 5 and 8 years, respectively. Limb salvage rate was 100%, 94% and 73% after 1 month, 5 and 8 years, respectively. CONCLUSIONS: The use of in situ venous allograft for the treatment of synthetic graft infection in the aorto-femoral segment has led to favourable short- and long-term results.

Arterial elasticity is associated with endothelial vasodilatory function and asymmetric dimethylarginine level in healthy subjects.

Arterial stiffening may be linked to the reduced bioactivity of nitric oxide (NO) and increased plasma concentrations of the endogenous NO synthase inhibitor asymmetric dimethylarginine (ADMA). The aim of this study was to investigate whether large (C1) and small artery (C2) elasticity is associated with endothelial function index (EFI) and plasma concentration of ADMA. We included 63 healthy subjects, aged 19 to 70 years, in the study. EFI, C1 and C2 were assessed by pulse wave analysis (PWA) and ADMA level was measured using an enzyme-linked immunoassay. Linear regression analysis revealed significant positive correlation between EFI and both C1 and C2 (R = 0.29, p = 0.02; R = 0.38, p = 0.002, respectively). A significant inverse association occurred between ADMA and C1 as well as C2 (R = -0.32, p = 0.03; R = -0.37, p = 0.009, respectively). In multiple regression analysis, C2 was determined by EFI, ADMA, age and BMI, and C1 was correlated with EFI, age and BMI. These findings suggest that endothelial vasodilatory dysfunction and accumulation of ADMA may be important mechanisms underlying reduced arterial elasticity in healthy subjects.

An antioxidant tetrapeptide UPF1 in rats has a neuroprotective effect in transient global brain ischemia.

Different glutathione analogues have potential to maintain or increase tissue glutathione level and to scavenge the reactive oxygen species. We designed and synthesized a novel non-toxic glutathione analogue, named UPF1, which possessed 60-fold higher hydroxyl radical scavenger efficiency in vitro, compared with glutathione itself, and investigated the effects of UPF1 on a four-vessel occlusion model of rats. The UPF1 was administered via the jugular vein in two separate experiments at two time points: 20 min before global brain ischemia and immediately before reperfusion. In both cases the number of pyramidal cells surviving in the subfield of CA1 at the dorsal hippocampus in the UPF1-treated groups of rats was twice as high as in the vehicle group.

Is elective abdominal aortic aneurysm repair accompanied by high grade oxidative stress?

BACKGROUND AND AIMS: During elective abdominal aortic aneurysm repair (AAAR), lower torso ischaemia-reperfusion event is unavoidable. Previous studies on AAAR have reported the importance of oxidative stress (OS) in ischaemia-reperfusion injury, however, the grade of OS has not been adequately clarified up to now. The aim of this study was to perform a complex investigation of the time-course and grade of systemic and cellular OS in patients undergoing AAAR. MATERIAL AND METHODS: Arterial blood samples were taken from 18 patients undergoing elective AAAR (at four points in time: before anaesthesia, 5 min after aortic clamping and 5 min and 30 min after clamp removal). Diene conjugates (DC), thiobarbituric acid reactive substances (TBARS), total antioxidative capacity (TAC), glutathione redox ratio (GSSG/GSH), and levels of antioxidative enzymes as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSHPx) were measured from the radial arterial blood. RESULTS: 30 min after the removal of the aortic cross-clamp, arterial CAT showed significant elevation (96.0 vs 56.9 U/l, p < 0.05); GSHPx was significantly elevated (51.5 vs 39.9 U/g Hgb, p < 0.05) and TAC was decreased (31.4 vs 36.5%, p < 0.05) in comparison with preoperative value. CONCLUSIONS: We found limited alterations of several OS parameters, which do not characterize either systemic or cellular high-grade OS during elective AAAR.

Exposure of rats to hyperoxia enhances relaxation of isolated aortic rings and reduces infarct size of isolated hearts.

Exposure of rats to hyperoxia before organ harvesting protected their isolated hearts against global ischaemia-reperfusion injury in a previous study. The present study investigates whether hyperoxia influences vasomotor function and regional ischaemia of the heart. Isolated rings of the thoracic aorta were obtained from rats immediately or 24 h after in vivo exposure to 60 min of hyperoxia (>95% O2), and the in vitro dose-response to phenylephrine (PHE), prostaglandin F2alpha (PGF2alpha) and endothelin-1 (ET-1), acetylcholine (Ach) and sodium nitroprusside (SNP) was assessed. Hyperoxia in vivo increased the relaxation of aortic rings to Ach and SNP, while it delayed contraction to PHE. The effect was more evident when the vessels were harvested immediately rather than 24 h after hyperoxic exposure. In separate experiments rat hearts were isolated immediately after hyperoxia, buffer-perfused, and subjected to 30 min of regional ischaemia and reperfused for 120 min. Infarct size was determined by triphenyl tetrazolium chloride staining. Hyperoxia significantly reduced infarct size. In normoxic controls 23.0 +/- 8.3% of the area at risk was infarcted, while in hyperoxic animals infarct size was 14.8 +/- 5.6% of the area at risk (P = 0.012). Exposure of rats to hyperoxia modifies the vasomotor response of isolated aortic rings, and reduces the infarct size of isolated rat heart. These novel aspects of hyperoxic treatment require further studies to explore the potential of its clinical application.

A novel mutation in the M1S1 gene responsible for gelatinous droplike corneal dystrophy.

PURPOSE: To identify the genetic defect in the M1S1 gene causing gelatinous droplike corneal dystrophy (GDLD) in an Estonian family. METHODS: DNA was extracted from members of a GDLD-affected family and control persons. Polymerase chain reaction followed by direct sequencing was used to detect mutations in the M1S1 gene. Sequencing results were confirmed with restriction analysis. RESULTS: Sequencing of the M1S1 gene revealed a novel mutation and a common polymorphism. All patients with GDLD were found to be homozygous for the insertion of nucleotide C in position 520 in M1S1. The mutation leads to formation of truncated protein. The mutation was excluded in 103 normal, unaffected individuals. Very close to the location where the mutation was identified in the M1S1 gene, a single-nucleotide polymorphism (518A/C) was found, changing aspartic acid to alanine at codon 173. CONCLUSIONS: The data indicate that mutation ins520C in the M1S1 gene is the primary cause of GDLD in the family studied.