Plasma Alkaline Phosphatase Activity Research Articles

Determination of plasma bone-specific alkaline phosphatase isoenzyme activity in Holstein calves using a commercial agarose gel electrophoresis kit.

We measured the bone-specific alkaline phosphatase (ALP) isoenzyme activity in 67 plasma samples from 14 newborn Holstein calves using both a conventional method (featuring heat inactivation) and a commercial agarose gel electrophoresis (AGE) kit; the relevant isoenzymes were termed bone-specific ALP (BAP) and ALP isoenzyme 3 (ALP3). We explored whether the AGE kit afforded reliable data when used to analyze samples from Holstein calves. The blood was collected from the jugular vein of each calf immediately prior to the first colostrum feeding (pre-feeding), 20 and 40 h after pre-feeding, and on days 4 and 7; whereas three samples (from three calves) were not obtained. The total plasma ALP activity varied widely, exceeding the ranges of reference values. On electrophoresis, 52 of 67 plasma samples (77.6 %) clearly contained both ALP isoenzyme 2 and ALP3, as did control human serum. The total ALP activity of the 52 samples ranged from 166-1989 U/L (median: 1013 U/L), whereas the values for the other 15 samples (22.4%) exhibiting abnormal isoenzyme fractionation ranged from 1014-5118 U/L (median: 1780 U/L). In the 52 plasma samples exhibiting clearly separated isoenzymes, ALP3 and BAP activities were strongly positively correlated as revealed by Deming regression (y = 0.93x + 22.6, p⟨0.0001) and Bland-Altman analysis (ALP3/BAP activities limit of agreement: -5.1%). Thus, the AGE kit yields useful information on newborn calves, and can replace the conventional method when the total plasma ALP activity is less than approximately 1000 U/L.

Optimum dietary fiber level could improve growth, plasma biochemical indexes and liver function of largemouth bass, Micropterus salmoides

An 8 week feeding trial was conducted to evaluate the effects of dietary fiber levels on growth, plasma biochemical parameters and liver function of juvenile largemouth bass. Five isonitrogenous (49% crude protein) and isolipidic (11% crude lipid) diets were formulated to contain 1.24%, 3.43%, 5.12%, 6.95% and 8.50% fiber levels, respectively. Four groups of largemouth bass were randomly assigned to each diet. The results indicated that largemouth bass fed the diets containing 8.5% fiber had significantly lower weight gain (WG) and protein efficiency ratio (PER) than those fed the other fiber diet. The whole-body lipid and liver lipid contents and hepatosomatic index (HSI) decreased with increasing dietary fiber levels，while whole-body protein content rose. Meanwhile, plasma triglyceride (TG) and total cholesterol (TC) contents and aspartate transaminase (AST) activity significantly decreased as dietary fiber levels increased, whereas plasma alkaline phosphatase (ALP) activity significantly rose. The carnitine palmitoyltransferase I (CPT-1), adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK) activities and peroxisome proliferator–activated receptor (PPAR-α) content in liver significantly increased with increasing dietary fiber levels. Furthermore, the activities of superoxide dismutase (SOD), catalase (CAT) and oxide synthase (NOS) increased and the malondialdehyde (MDA) content in liver decreased significantly as dietary fiber levels increased. In addition, the liver histological investigation showed an improvement in liver histostructure at high fiber levels. These findings indicated that dietary fiber levels could improve liver function and antioxidant ability of M.salmoides; also, higher dietary fiber levels have a positive impact and decrease the potential risk of disease in M. salmoides. Based on WG, a dietary fiber level of 4.07% was optimal for growth performance of juvenile largemouth bass.

2066 Much Ado About Nothing: Isolated Raised ALP in IBD Patients Due to Macrocomplex

INTRODUCTION: Macro-Alkaline Phosphatase (ALP) is a rare benign condition characterised by increased plasma ALP activity due to an immunoglobulin-bound form of ALP. Due to an autoimmune process, antibodies can be generated against any of the enzymes, leading to the formation of “macroenzymes”, which interferes with the correct laboratory measurement of the native enzymes. These have been identified for several enzymes including alkaline phosphatase (ALP), amylase, AST, creatine kinase, γ-glutamyl transferase and lactate dehydrogenase. CASE DESCRIPTION/METHODS: 51-year-old woman with a long-standing history of ulcerative colitis and psoriasis presented for establishing care. She reported alternating diarrhea and constipation and musculoskeletal pains but no jaundice or itching. Her labwork showed normal levels of AST, ALT and total bilirubin but raised ALP (190 units/L, UNL-103). Hepatic origin of raised ALP was confirmed with mildly elevated GGT (42 U/L, UNL- 37). Repeat labs done at monthly intervals showed persistently elevated ALP ranging between 140-240 U/L. She underwent extensive workup to rule out an obstructive pathology, including a USG and MRCP which were unremarkable and liver biopsy which showed preserved hepatic architecture with minimal steatosis. Fractionation of her ALP via gel electrophoresis showed Macro-ALP complexes. She was followed up for a period of 2 years, with persistently elevated ALP but no obstructive symptoms or liver dysfunction. DISCUSSION: Isolated raised ALP is a common finding seen in up to 16% of hospitalized patient. However, this becomes concerning in patients with IBD due to its association with disorders of the hepatobiliary system, particularly Primary Sclerosing Cholangitis, seen in upto 5% of IBD patients The mean time from the development of symptoms of IBD to the diagnosis of PSC was 2.3 months. The biochemical parameters are consistent with cholestasis. Our patient was beyond the time frame for the development of PSC and showed no symptoms of signs of cholestasis. Macro-ALP as a cause of persistently elevated ALP has been reported twice earlier in literature. Strategies to identify macro enzymes include polyethylene glycol precipitation,counter-immunoelectrophoresis or gel electrophoresis. In general, the identification of macro-enzyme complexes is important to rule out pathological causes of raised serum enzyme activity and avoid unnecessary and potentially harmful diagnostic investigations.

Physiological Responses of Pacific White Shrimp Litopenaeus vannamei to Temperature Fluctuation in Low-Salinity Water.

Temperature is a significant environmental factor in aquaculture. To investigate the physiological responses during temperature fluctuation (28~13°C), experimental shrimps (Litopenaeus vannamei) were treated with gradual cooling from acclimation temperature (AT, 28°C) to 13°C with a cooling rate of 7.5°C/day and rose back to 28°C at the same rate after 13°C for 24 h. Hepatopancreas histological changes, plasma metabolites concentrations, relative mRNA expression of unfolded protein response (UPR) pathway and apoptosis in hepatopancreas and hemocyte were investigated. The results showed that with the decline of temperature, the number and volume of the secretory cells in hepatopancreas increased significantly, the tubule lumen appeared dilatated, and the epithelial cell layer became thinner. The contents of glucose (Glu) significantly decreased to the minimum value of 13°C for 24 h. The contents of triglyceride (TG), total cholesterol (TC), and total protein (TP) increased and reached the peak of 13°C for 24 h. Alkaline phosphatase (ALP) and alanine aminotransferase (ALT) activities in plasma reached the lowest and highest value in 13°C, respectively. The expressions of all genes related to UPR and apoptosis in the hepatopancreas and hemocytes were significantly changed during the cooling process and reached the highest level of 13 and 13°C for 24 h, respectively. During re-warming stage, the histopathological symptoms got remission and each of the plasma metabolite concentrations and gene expressions returned to AT levels. These results revealed that pacific white shrimp can adapt to a certain level of temperature fluctuation by self-regulation.

Evaluation of immune response and performance of silver catfish fed functional linseed fibres in response to hypoxia stress

This study was conducted to evaluate the immunostimulating activity and growth promoting of diets supplemented with different rations of soluble and insoluble linseed fibre to Rhamdia quelen under hypoxia-induced acute stress. For this reason, soluble and insoluble fractions of linseed fibre were concentrated separately and combined into four ratios (1:0.5; 1:1; 1:2 and 1:4), which were added to the diets of silver catfish (6.43 ± 0.12 g) and evaluated in a biological assay, along with a control diet (without addition of linseed fibre). After being fed the experimental diets for 45 days, specimens of silver catfish were submitted to hypoxia-induced acute stress. They were kept out of water for 60 s. Immediately afterwards, blood and cutaneous mucus were collected for subsequent determination of immunological indicators and stress. After stress, the fish were weighed and measured for performance parameter calculation. The experimental design was completely randomized with five treatments and four replications. The data underwent analysis of variance and the means were compared by Tukey's test (p < .05). The fish fed diets containing the 1:2 and 1:4 soluble:insoluble fibre ratios, showed higher total protein content, globulin, plasma alkaline phosphatase activity, and performance in addition to higher mucoprotein content in the cutaneous mucus of the fish. Regardless of their ratio in the diet, linseed fibre provided higher plasma levels of total immunoglobulins and reduction in cortisol levels. The 1:1, 1:2 and 1:4 diets led to higher levels of total immunoglobulins and alkaline phosphatase activity in cutaneous mucus. The results indicate that linseed fibre has a stress- reduction, immunostimulant and a growth promoter effect on silver catfish. The 1:2 or 1:4 soluble: insoluble fibre ratios provided greater stimulation of the target immunological indicators and performance.

Grapefruit juice exerts anti-osteoporotic activities in a prednisolone-induced osteoporosis rat femoral fracture model, possibly via the RANKL/OPG axis.

This study aimed to shed light on the protective and therapeutic anti-osteoporotic effects and mechanisms of action of grapefruit juice (GFJ) on prednisolone-induced osteoporosis a rat femoral fracture model. We found that treating rats with GFJ before and/or after prednisolone-induced osteoporosis resulted in increased bone density, total mineral content, and calcium content to counteract the osteoporotic effects of prednisolone. In parallel, the histological and ultrastructural results of the GFJ-treated groups correlated well with enhanced breaking strength of femurs subjected to a constant load. Furthermore, GFJ treatment before and after prednisolone-induced osteoporosis decreased plasma alkaline phosphatase and tartrate-resistant acid phosphatase activities and increased the level of insulin-like growth factor 1. Mechanistically, our immunohistochemistry study showed that GFJ ameliorated prednisolone-induced osteocalcin depletion, decreased receptor activator of nuclear factor kappa-B ligand (RANKL) expression, and increased osteoprotegerin (OPG) expression. GFJ showed a beneficial anti-osteoporotic effect against prednisolone-induced osteoporosis in rats, possibly via the RANKL/OPG axis, suggesting that GFJ might be a good candidate for developing anti-osteoporotic drugs.

Influence of bone meal degelatinisation and calcium source and particle size on broiler performance, bone characteristics and digestive and plasma alkaline phosphatase activity

ABSTRACT 1. The current experiment was performed to elucidate the effects of degelatinised bone meal (DBM) in combination with different particle sizes of limestone or oyster shell on broiler performance, bone characteristics and digestive and plasma alkaline phosphatase (ALP) activity. 2. Treatments were applied as a 3 × 3 factorial arrangement with three sources of P (DCP, bone meal and DBM) and three particle sizes (50, 100 and 200 µm) of limestone. Chickens were given either DCP or DBM with oyster shell (523 µm), resulting in a total of 11 treatments with 5 replicates of 8 chicks. 3. Performance criteria were measured weekly. Tibia strength, ash, calcium (Ca) and phosphorus (P) content and plasma P and Ca concentration along with plasma and intestinal alkaline phosphatase (ALP) activity and P digestibility were measured on d 14 and 28. 4. Body weight and FCR were improved in chicks which were fed DBM or oyster shell in comparison to the DCP and limestone respectively (P ≤ 0.05). Performance was influenced (P ≤ 0.05) by particle size; with coarser particles BW and feed intake were increased (P ≤ 0.05). Tibia shear force and P content were reduced (P ≤ 0.001), whereas tibia shear energy, length, ash and Ca content were increased by substitution of DCP with DBM or bone meal (P ≤ 0.001; P ≤ 0.05). A significant difference was observed in the tibia length between the chicks fed oyster shell or limestone with different particles (P ≤ 0.05). Plasma P concentration was reduced in chicks were fed with DBM, bone meal and lower limestone particle size. Intestinal ALP activity was increased (P ≤ 0.001) in chicks which were fed DBM, bone meal, oyster shell or coarse particles of limestone. The P digestibility in chicks fed bone meal was lower than that of those fed DBM or DCP (P ≤ 0.01). Overall, gelatin removal from bone meal improved broiler bone characteristics through the P digestibility and intestinal ALP activity enhancement.

The Effect of Artemisinin-based Combination Therapy (ACT) Antimalaria Drugs on Liver Enzymes in Pregnancy

Aims: Artemisinin-based Combination Therapies (ACTs) are employed as first-line agents in malaria chemotherapy. This study is aimed at assessing the effects of ACTs on renal function of pregnant women.&#x0D; Study Design: Comparative study.&#x0D; Place and Duration of Study: Pregnant women aged 18 to 50 years were recruited from antenatal clinic of Obstetrics and Gynecology Department of Ekiti State Hospital, Ado Ekiti, Nigeria between 2016 and 2018&#x0D; Methodology: One hundred and eighty pregnant women were grouped into three which include: Sixty pregnant women with malaria parasite on ACT drugs (Group A), sixty pregnant women with malaria parasite not on ACT drugs (Group B), sixty pregnant women without malaria parasite (Group C/control). Plasma Aspartate Aminotransferase (AST), Alkaline Phosphatase (ALP), Alanine Transaminase (ALT) and Lactate Dehydrogenase (LDH) activities were evaluated by standard methods. The data collected were analyzed using one-way analysis of variance (ANOVA) and Student’s t test to compare the data between the test groups and control.&#x0D; Results: Results showed there was a significant decrease (P=.05) in Body Mass Index in the pregnant women with malaria on ACT and those that were not on ACT when compared with control (24.1± 0.32 versus (25+ 2.30 vs 27± 1.62). A significant increase (P=.05) occurred in the levels of ALP,AST, ALT and LDH in pregnant women with malaria not on ACT drugs when compared the control (168.45±0.19, 10.0±0.27, 8.19±0.25, 4.5±0.21) versus (143.20±0.12, 8.71±0.30, 5.99±0.21, 2.08±0.19),while no significant difference occurred in the levels of ALP,AST, ALT and LDH in pregnant women with malaria on ACT when compared with control (141.60±0.78, 8.02±0.32, 6.10±0.30, 2.75+ 0.20) vs (143.20±0.12, 8.71±0.30, 5.99±0.21, 2.08±0.19).&#x0D; Conclusion: Normal therapeutic dose of ACT has no harmful effect on the liver in pregnancy.

Time – Course of Sodium Arsenate Induced Hepatotoxicity and Nephrotoxicity in Male Wistar Rats

Arsenic exposure has been implicated by several epidemiological studies as an important metalloid that is currently poisoning millions of people globally. In order to investigate the time – course of arsenic exposure on hepatic and renal toxicity, male albino rats (n=45) were exposed to arsenic (100 ppm, 150 ppm and 200 ppm) for 4, 8 and 12 weeks as sodium arsenate in their drinking water. Control animals (n=15) received distilled water for the same period after which blood and vital organs were removed from the animals and analyzed for alanine amino transaminase (ALT), aspartate amino transaminase (AST) gamma amino transaminase (γGT), alkaline phosphatase (ALP), creatinine and urea spectrophotometrically. Histological changes in hepatocytes was also examined. Before the commencement of arsenic exposure, five animals were sacrificed to obtain baseline data. Significant elevation in plasma ALT, AST, γGT and alkaline phosphatase activities characterized the effect of the arsenical at all doses and time interval relative to the controls. Plasma levels of creatinine and urea were also elevated at all-time intervals in the arsenic group. In most of the cases observed, the elevated level of these biochemical marker in circulation are time – and dose – dose dependent. Hepatic histopathology reveals degeneration of cytoplasmic contents, evidence of necrosis, collapse of central vein, cytoplasmic inclusion and enlarged hepatic sinusoids in arsenic – exposed groups. These findings suggest that different dose regimens of sodium arsenate at different time interval caused degenerative changes in hepatic and renal tissues in rats in dose – and time – dependent fashion. Keywords: Arsenic, Time – course, Hepatotoxicity, Nephrotoxicity DOI: 10.7176/JNSR/9-4-09

Protective effect of crocin on food azo dye tartrazine-induced hepatic damage by improving biochemical parameters and oxidative stress biomarkers in rats.

The objective of the present study was to demonstrate the protective effect of crocin on the adverse effects of tartrazine on liver. Crocin is a carotenoid and a strong free radical scavenger. Forty rats were randomly divided into 4 groups (n = 10). The first group was the control group (C) and saline solution was administered to this group. The second group (Cr) was administered 50 mg/kg crocin. The third group (T) was administered 500 mg/kg tartrazine. The fourth group (T+Cr) was administered the same doses of both crocin and tartrazine as the previous groups for 21 days. It was determined that tartrazine increased liver superoxide dismutase (SOD) activity, malondialdehyde (MDA) and total oxidant status (TOS) levels and catalase (CAT) activity, decreased glutathione (GSH), and total antioxidant status (TAS) levels. Furthermore, tartrazine administration resulted in significant increases in plasma aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activities and pathological changes in the liver. When tartrazine administered rats were treated with crocin for 21 days, the biochemical parameters improved, and liver tissues were restored. Thus, it was demonstrated that crocin had protective effects on the adverse effects caused by tartrazine administration.

Effect of lesimarin against thioacetamide-induced liver cirrhosis in rat

Liver cirrhosis is one of chronic liver diseases with high disability and mortality accompanying fibrosis, hepatocyte damage, and liver dysfunction. In this study, the hepatoprotective and the antifibrotic properties of lesimarin(lecithin - silymarin - Artemisia messerschmitiana Besser(AMB) extract complex at 11:3:6 ratio) on rat hepatic fibrosis induced by thioacetamide (TAA) was investigated. Rats were divided into seven groups: control, lesimarin, TAA, TAA+lesimarin, TAA+lecithin, TAA+silymarin, TAA+AMB. Rats were administered with TAA at a dose of 200 mg/kg body weight intraperitoneally twice a week for three months. Lesimarin, lecithin, silymarin and AMB were administered at a dose of 1.0, 1.0, 0.5, 1.0g/kg body weight orally daily for three months, respectively. TAA administration resulted in hepatic fibrosis, significant decrease in body weight, albumin level and A/G ratio and increase in plasma transaminase, GGT(γ-glutamyltransferase) and ALP(alkaline phosphatase) activities as well as hepatic hydroxyproline content, which were attenuated by lesimarin administration. Lesimarin was found to decrease AST, ALT and GGT, ALP and bilirubin, hydroxyproline levels and increase albumin level and A/G ratio and its effect is more prominent than those of individual constituents. These results suggest this new drug, lesimarin, might be a promising drug to be used for chronic liver diseases.

Evaluation of Serum Status of Biochemical Indices of Liver Injury and Oxidative Stress in Rats Exposed to Warri River Levels of Pb and Other Identified Metallic Co Pollutants

This study investigated serum status of biochemical indicators of liver injury and oxidative stress in rats exposed to Warri River level of lead(Pb) alone and in the presence of metallic co-pollutants. A total of 55 albino rats (of Wistar strain) weighing an average of 150.00± 09.00g, divided into 11 groups were used for the study. Groups I and II represented the deionized and Pti borehole water controls, while groups III- XI represented the test rat groups orally treated with water containing laboratory reconstituted Warri River Pb level on one hand and in the presence of laboratory reconstituted identified metallic co-pollutants including Fe, Ca, Cu, Mn, Mg, Zn via water on the other hand. The serum biochemical –hepatotoxic indices investigated were liver/body wt. ratios, body wt. change, lipid per oxidation products, plasma ALT and AST, plasma and liver alkaline phosphatase activities, plasma catalase and superoxide dismutase activities, plasma total and conjugated bilirubin level, plasma and urine glucose concentration, and plasma and urine total protein concentration. Our findings revealed an overall significant (P&lt;0.05) decrease in liver/body wt ratios and body wt change , significant (P&lt;0.05) increase in plasma ALT and AST activities, induced ALP and ACP activities, increase in SOD and catalase activities, increased plasma and urine bilirubin concentrations, decreased plasma and increased urine total protein concentrations, increased Malondialdehyde (MDA) levels, while plasma and urine glucose levels were elevated in the groups of rats exposed to Pb only, Pb + Cu, Pb + Fe and Pb + Zn, Pb + All metallic co-pollutants, and river water relative to their respective controls (deionized water and Pti tap water groups). There was a significant (P&lt;0.05) reversal of the above parameters in the groups of rats exposed to Pb + Ca, Pb + Mn, Pb + Mg. There was also a difference in liver/weight ratio, body wt. change and all the other parameters evaluated in this study, between groups of rats treated with Warri river water relative to the laboratory reconstituted water, although the changes were not significant (P&gt;0.05). Our findings revealed that, the presence of Ca, Mg and Mn in the river water significantly (P&lt;0.05)) reversed the induced activities of ALT, AST, ACP and ALP by Pb and some identified metallic co pollutants like Cu, Fe and Zn. This study also revealed the possibility of significant (P&lt;0.05) decrease in the activities of superoxide dismutase (SOD), catalase , plasma total and direct bilirubin and lipid per oxidation products of rats exposed to Warri River level of Pb in the presence of Ca, Mg and Mn relative to the Pb only group.

Effects of gelatin prepared from calf bones rich in phosphorus on broiler performance, bone characteristics and digestive enzymes activity

ABSTRACT1. Gelatin prepared from calf bones (GCB) is a novel source of high-quality protein and phosphorus. Its inclusion in broiler chicken diets may improve bone strength, plasma and digestive alkaline phosphatase activity (ALP), phosphorus digestibility and performance of broilers. Therefore, di-calcium phosphate in a corn-soy control diet was replaced with 12, 24, and 36 g/kg of GCB in a completely randomised design with four treatments of six replicates and 10 chicks in each pen. The trial lasted from 1 to 42 d of age.2. Body weight and feed intake were measured weekly. Plasma calcium and phosphorus concentration along with plasma and digestive ALP were assayed throughout the trial. Trypsin, α-amylase, lipase and total protease activity were assayed at 14 and 28 d of age. Tibia ash, calcium and phosphorus content and breaking strength were measured at 14, 28 and 42 d of age. Phosphorus digestibility was measured at 36 d of age.3. Body weight and feed intake showed no significant differences between controls and diets containing 12 and 36 g/kg GCB. Tibia ash and tibia length were increased by supplementation of GCB (P ≤ 0.001). Tibia calcium and phosphorus content were increased by GCB inclusion at 14 d of age (P ≤ 0.001). Digestive alkaline phosphatase activity was increased and trypsin activity was reduced by inclusion of GCB (P ≤ 0.001; P ≤ 0.004). α-amylase activity decreased by inclusion of 12 and 24 g/kg GCB, whereas an increase in α-amylase activity was observed by inclusion of 36 g/kg GCB (P ≤ 0.001). Supplementation of diets with GCB increased phosphorus digestibility (P ≤ 0.01) and suppressed ileum growth during the experimental period.4. Results of the current study showed that phosphorus from gelatin can greatly improve broiler bone characteristics and phosphorus digestibility and complete elimination of inorganic phosphate sources from broiler diets is feasible with inclusion of 36 g/kg high phosphorus gelatin.

Long-term induced hyperthyroidism in breeder hens: Effects on blood plasma biochemical attributes, indicators of oxidative stress, and markers of liver function

A 4-week-long administration of extra thyroxine in broiler breeder hens was suggested to reduce the cold-induced ascites in their offspring. In the present study the hypothesis was tested to disclose the plausible adverse effects of long-term maternal hyperthyroidism (100 days) on blood plasma biochemical attributes, indicators of oxidative stress, and markers of liver function in the exposed breeder hens. Also, the association between egg production rate and egg shell strength with alkaline phosphatase activity of plasma were tested. Seventy 47-week-old Cobb 500 breeder hens were randomly allotted to two treatment groups as control (CON) and hyperthyroid (HYPER), each consisting of five replicates of seven birds. Thyroxine (T4; 0.3 mg/bird/d) was orally administered to the HYPER group for 100 consecutive days; whereas the CON group received the drinking water only. Blood sampling was done every 5 week from 47 to 64 week of age for quantification of plasma T4 and T3, and biochemical attributes as well as indicators of oxidative stress. Results indicated that plasma concentration of T4 was greater for the HYPER birds (P 〈 0.05); however, that of T3 was not different between the experimental groups. Total cholesterol, HDL, LDL, and VLDL, and triglyceride concentrations were not affected by T4 administration. Induced hyperthyroidism had no apparent effect on in vitro plasma lipoperoxidation indices, including CDMax, CDAUC, and CD lag phase; whereas, CD oxidation rate and MDA were greater in HYPER group as compared with their CON counterparts. The effect of oral T4 on plasma aspartate aminotransferase and alanine aminotransferase was not significant although the plasma level alkaline phosphatase was higher in HYPER birds (P > 0.05). No significant treatment effect was observed on egg production. Hens belonging to HYPER group recorded a higher egg shell thickness than those of the CON birds, although the correlation coefficient of egg shell thickness with plasma concentration of alkaline phosphatase was not significant. Overall, apart from the changes in parameters evaluated, the values were within their biological ranges. Therefore, it might be stated that the long term administration of T4 did not adversely affect the biochemical characteristics of broiler breeder hens. Future studies are needed to make a final decision on use of this treatment to reduce the ascites incidence in offspring.

Dietary yeast culture modulates immune response related to TLR2-MyD88-NF-kβ signaling pathway, antioxidant capability and disease resistance against Aeromonas hydrophila for Ussuri catfish (Pseudobagrus ussuriensis)

The aim of the present study was to investigate effects of dietary yeast culture on immune response related to TLR2-MyD88-NF-kβ signaling pathway, antioxidant capability and disease resistance against Aeromonas hydrophila for Ussuri catfish (Pseudobagrus ussuriensis). A total of 240 Ussuri catfish (mean weight of 7.39 ± 0.32 g) were randomly distributed into four groups that fed diets containing 0 (Y0), 10 (Y1), 20 (Y2) and 30 (Y3) g kg−1 yeast culture for 8 weeks. The results indicated that dietary 10 g kg−1 yeast culture supplementation significantly down-regulated mRNA levels of TLR2, MyD88, NF-kβ p65, IL-1β and IL-8 in the liver tissue compared with the control group (P < 0.05). Simultaneously, serum lysozyme (LZM) activity, respiratory burst activity (RBA) of phagocytes, plasma alkaline phosphatase (AKP) activity and immunoglobulin M (IgM) content were significantly improved in fish fed Y1 diet (P < 0.05). Fish fed Y1 diet had significantly higher serum alternative complement pathway activity (ACH50) and plasma complement 3 (C3) content than the Y3 group (P < 0.05). However, no significant differences were observed in plasma acid phosphatase (ACP) activity and complement 4 (C4) content among the groups (P > 0.05). Fish cumulative mortality rate (CMR) in the Y1 and Y2 groups were significantly lower than that in Y0 and Y3 groups (P < 0.05), and the lowest CMR was observed in the Y1 group after challenge by A. hydrophila. The highest hepatic superoxide dismutase and glutathione peroxidase activities, total antioxidant capacity and the lowest malondialdehyde content were found in Y1 group, but no significant difference was found in hepatic catalase activity among the groups (P > 0.05). These results demonstrate that dietary 10 g kg−1 yeast culture could effectively improve the immunity, antioxidant capability and disease resistance against A. hydrophila for Ussuri catfish and could down-regulate the mRNA expression levels of pro-inflammatory cytokines modulated by TLR2-MyD88-NF-kβ signaling pathway.

Sensitive and selective colorimetric detection of alkaline phosphatase activity based on phosphate anion-quenched oxidase-mimicking activity of Ce(Ⅳ) ions

As alkaline phosphatase (ALP) plays crucial roles in disease warning and dephosphorylation-related cellular regulation, it is widely recognized as an important biomarker for clinical diagnosis. In this work, we proposed a facile colorimetric assay based on phosphate anion-quenched oxidase-mimicking activity of Ce(Ⅳ) ions for sensitive and selective detection of ALP activity. Free Ce(Ⅳ) ions exhibited a strong oxidase-like capability (providing a 40-fold catalytic turnover number compared with CeO2) to catalyze the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) into its blue product TMBox mediated by dissolved O2 at neutral pH, thus triggering a remarkable color reaction visually. When PO43− was added, its strong affinity to Ce(Ⅳ) ions rapidly precipitated the free Ce(Ⅳ) ions, resulting in the quenching of their enzymatic ability. Given that ALP catalyzed the hydrolysis of adenosine triphosphate to produce PO43−, determination of ALP activity could be achieved using the colorimetric assay with no need of complicated instrumentation and protocol. As demonstrated, our assay offered a highly sensitive readout for ALP activity in two linear scopes of 0–50 U L−1 and 50–250 U L−1, providing a detection limit down to 2.3 U L−1. Besides, it could provide specific response toward ALP against other enzymes and biological species. Furthermore, the developed assay was successfully applied to evaluate ALP activity in human plasma accurately and reliably, indicating its great promise as a powerful and convenient tool for monitoring of ALP activity in clinical practice.

Changes in Plasma Metabolites Concentrations in Obese Dogs Supplemented With Anti-oxidant Compound.

The aim of this study is to discuss the effect of anti-oxidant supplement (Rv-PEM01-99, Kibun Foods, Inc., Tokyo, Japan) on changes in energy metabolism in obese dogs. 200 mg/kg/day of Rv-PEM01-99 (equivalent to 5 mg kg/day of quercetin derivative) were applied for 6 weeks to the Beagle dogs fed high fat diet (HFD) or control diet (CD). In the present study, body weight (BW) decreasing effect of Rv-PEM 01-99 in obese dogs was not clear. However, plasma alkaline phosphatase (ALP) activities at the end of experiment were significantly decreased compared to those at the start of experiment in obese dogs supplemented with Rv-PEM 01-99 (paired-t test, p < 0.05). In control dogs supplemented with Rv-PEM 01-99, Plasma malondialdehyde (MDA), and triglycerides (TG) levels and lactate dehydrogenase (LDH) activities were significantly decreased compared to those at the start of experiment (paired-t test, p < 0.05). From these findings, Rv-PEM 01-99 seems to be not harmful for dogs. Anti-lipid peroxide effect and liver function improvement are expected in the dogs supplemented with Rv-PEM 01-99.

Effects of dietary arginine on intestinal antioxidant status and immunity involved in Nrf2 and NF-κB signaling pathway in juvenile blunt snout bream, Megalobrama amblycephala

The present study assessed the effects of dietary arginine on intestinal antioxidant status and immunity involved in Nrf2 and NF-κB signaling pathway in juvenile blunt snout bream. Fish were fed three practical diets with graded arginine levels (0.87%, 1.62% and 2.70%) for 8 weeks. Compared with the control group (0.87%), the counts of white blood cell (WBC), red blood cell (RBC) and hemoglobin (HGB) content were significantly improved at dietary arginine levels of 1.62% (P<0.05). Plasma albumin (ALB) levels and alkaline phosphatase (ALP) activities were significantly improved at dietary arginine levels of 1.62% and 2.70% (P < 0.05). Alanine transaminase (ALT) activity was decreased in fish fed with 1.62% dietary arginine level (P<0.05). Plasma glutathione peroxidase (GPx) activities, copper-zinc superoxide dismutase (Cu/Zn-SOD) activities, total antioxidant capacity (T-AOC) activities and glutathione (GSH) levels were significantly increased at dietary arginine levels of 1.62% and 2.70% (P<0.05). Plasma total superoxide dismutase (T-SOD) activities and catalase (CAT) activities were significantly improved in fish fed with 1.62% dietary arginine level. Significantly higher manganese superoxide dismutase (Mn-SOD) activity was observed in fish fed with 1.62% dietary arginine level compared with 2.70% dietary arginine level (P<0.05). 1.62% and 2.70% dietary arginine levels significantly lowered malondialdehyde (MDA) levels. The relative expression of nuclear factor erythroid 2-related factor 2 (Nrf2) was significantly increased in fish fed with 1.62% dietary arginine level, inversely, the relative expression of Kelch-like ECH-associated protein 1 (Keap1) showed a converse trend. 1.62% and 2.70% dietary arginine levels significantly improved the relative expressions of Cu/Zn-SOD, GPx and CAT. Furthermore, 2.70% dietary arginine level significantly lowered the relative expression of Mn-SOD compared with the control group and 1.62% dietary arginine levels. The relative expressions of Interleukin 1β (IL-1β), tumour necrosis factor-α (TNF-α) and nuclear factor-kappa B (NF-κB) were lowered in fish fed with 1.62% dietary arginine level. 1.62% and 2.70% dietary arginine levels significantly improved the relative expressions of transforming growth factor-β (TGF-β). Hematocrit (HCT), aspartate aminotransferase (AST) activities, interleukin 8 (IL-8) and interleukin 10 (IL-10) expressions were not significantly affected by the graded dietary arginine levels. These results suggest that the optimal dietary arginine level plays an important role in enhancing antioxidant and immune status to maintain the intestinal health of juvenile blunt snout bream.

Hepatotoxicity of vanadyl sulfate in nondiabetic and streptozotocin-induced diabetic rats.

This study examined the effects of vanadyl sulfate (VOSO4) on the livers of nondiabetic and streptozotocin-induced diabetic rats. Rats were divided into 6 groups. Groups 1, 2, and 3 consisted of nondiabetic rats that were, respectively, control animals or those receiving an intraperitoneal (i.p.) injection of either 5 or 10 mg·kg-1 (i.p.) VOSO4 for 30 days. Groups 4, 5, and 6 consisted of diabetic animals that were, respectively, control animals or those treated with 5 or 10 mg·kg-1 (i.p.) VOSO4 for 30 days. Results showed that VOSO4 reduced body mass in nondiabetic rats, whereas it increased body mass in diabetic groups. Plasma transaminases (aspartate aminotransferase, alanine aminotransferase), lactate dehydrogenase, and alkaline phosphatase activities and malondialdehyde levels were increased, while liver catalase and superoxide dismutase activities were profoundly decreased in diabetic animals in comparison with enzyme activities in the nondiabetic group. Rats in the diabetic group also showed notable oxidative damage to the liver. Treatment of diabetic rats with VOSO4 decreased the hepatotoxic markers, significantly restored the activities of antioxidant enzymes, and attenuated histopathological changes in liver tissue. In nondiabetic rats, VOSO4 treatment increased most of the hepatotoxic markers, reduced antioxidant enzyme activities, and induced pronounced oxidative damage in liver tissue. These data suggest that treatment with VOSO4 exerts toxic effects in healthy animals and significantly prevents liver oxidative damage in streptozotocin-induced diabetic rats, but without total safety. Further studies are needed to clarify its mechanism of action.

Hydrolysis of Extracellular Pyrophosphate increases in post-hemodialysis plasma

Vascular calcification (VC) is associated with significant morbidity and mortality of dialysis patients. Previous studies showed an association between loss of plasma pyrophosphate and VC. Moreover, loss of pyrophosphate occurs during dialysis in this population, suggesting that therapeutic approaches that prevent reduction of plasma pyrophosphate levels during dialysis could improve the quality of life of dialysis patients. This study found that pyrophosphate hydrolysis was 51% higher in post- than pre-dialysis plasma. Dialysis sessions modified the kinetic behavior of alkaline phosphatase, increasing its Vmax and reducing its Km, probably due to the elimination of uremic toxins during dialysis. At least 75% of alkaline phosphatase activity in human plasma was found to depend on a levamisole-sensitive enzyme probably corresponding to tissue non-specific alkaline phosphatase (TNAP). Dialysis increased total plasma protein concentration by 14% and reduced TNAP enzyme by 20%, resulting in an underestimation of pyrophosphate hydrolysis in post-dialysis plasma. Levamisole inhibited TNAP activity (IC50, 7.2 µmol/L), reducing pyrophosphate hydrolysis in plasma and increasing plasma pyrophosphate availability. Alkaline phosphatase is also found in many tissues and cells types; therefore, our results in plasma may be indicative of changes in phosphatase activity in other locations that collectively could contribute significantly to pyrophosphate hydrolysis in vivo. In conclusion, these findings demonstrate that dialysis increases pyrophosphate hydrolysis, which, taken together with previously reported increases in alkalization and calcium ion levels in post-dialysis plasma, causes VC and could be prevented by adding calcification inhibitors during dialysis.