Biomedical subjects

W Bottje

Publications and source records attributed to W Bottje.

16 recordsLinked to original sources

PubMed2006

Feed efficiency and mitochondrial function.

Studies have been conducted in our laboratory to assess differences in mitochondrial function and biochemistry in male broilers with high and low feed efficiency (FE) from the same genetic line and fed the same diet. Mitochondria obtained from broilers with low FE exhibited greater uncoupling of the electron transport chain (ETC) that was apparently due to site-specific defects in electron transport resulting in higher amounts of reactive oxygen species (ROS) compared with high FE mitochondria. Higher amounts of ROS production in Low FE mitochondria were likely responsible for higher protein carbonyl levels, indicative of higher protein oxidation compared with High FE mitochondria and tissue. In turn, higher protein damage in Low FE mitochondria may have contributed to lower activity of electron transport chain complexes relative to values observed in high FE mitochondria. Low FE mitochondria did not exhibit a compromised ability to carryout oxidative phosphorylation, and although there were differences in expression of certain electron transport chain proteins, there was nothing that would indicate that differences in coupling and respiratory chain activity could be due to a general decrease in protein expression between low and high FE mitochondria. The results of these studies provide insight into understanding cellular mechanisms associated with the phenotypic expression of feed efficiency in broilers.

Animal Feed↗

PubMed2006

Differential expression of mitochondrial and extramitochondrial proteins in lymphocytes of male broilers with low and high feed efficiency.

Studies were conducted to investigate relationships between mitochondrial and extramitochondrial protein expression, and protein oxidation in lymphocytes obtained from broilers in which individual feed efficiencies were obtained. Lymphocytes were isolated from male broilers from a single line that were shown to exhibit either low (0.48 +/- 0.02, n = 8) or high (0.68 +/- 0.01, n = 7) feed efficiency (FE). Western blot analysis showed that, compared with lymphocytes from high FE broilers, lymphocytes from low FE broilers exhibited a) higher amounts of oxidized proteins (protein carbonyls), b) lower amounts of 3 mitochondrial proteins [core I, cyt c 1 (complex III), and ATP synthase (complex V)], and c) higher amounts of 2 proteins [30 S (complex II) and COX II (complex IV)]. Two-dimensional gel electrophoresis revealed that the intensities of 25 protein spots from pooled samples of lymphocytes from high and low FE broilers differed by 5-fold or more. Three of these protein spots were picked from the gel and subjected to matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry analysis. One protein spot of ~33 kDa was tentatively identified by MALDI-TOF as a fragment of collapsin-2, a component of semaphorin 3D. The results of this study provide further evidence of increased oxidation associated with low FE and further evidence of differential protein expression associated with the phenotypic expression of feed efficiency.

Animal Feed↗

PubMed2005

Glutathione and respiratory chain complex activity in duodenal mitochondria of broilers with low and high feed efficiency.

We previously observed increased reactive oxygen species (ROS) production in intestinal mitochondria obtained from broiler breeder males with low feed efficiency (FE, gain-to-feed). Because antioxidants are critical for combating ROS-mediated oxidative stress and preserving mitochondrial function, the objectives of this study were 1) to determine levels of reduced glutathione (GSH), a major antioxidant in mitochondria, 2) to measure activities of GSH recycling enzymes: GSH peroxidase and GSH reductase, and 3) to establish relationships between antioxidants and respiratory chain complex activities (complexes I, II, III, IV, and V) in broiler breeder males with low and high FE. Duodenal mitochondria were isolated from broilers with low (0.62 +/- 0.01, n = 8) and high (0.80 +/- 0.01, n = 8) FE. Activities of respiratory chain complexes, GSH peroxidase, and GSH reductase, and levels of GSH were measured by UV spectrophotometry. There were no differences in GSH peroxidase or reductase activities or in individual complex activities between groups but GSH levels tended to be higher (P = 0.075) and oxidized to reduced glutathione ratio tended to be lower (P = 0.077) in broilers with high FE. Regression analysis revealed significant correlations (P < or = 0.05) between mitochondrial GSH and activities of complexes II, IV, and V with R2 values of 0.35, 0.56, and 0.49, respectively. These data suggest that GSH may be important in maintaining or enhancing the activity of certain respiratory chain complexes and could be involved in the phenotypic expression of feed efficiency in broilers.

Animals↗

PubMed2005

Compromised liver mitochondrial function and complex activity in low feed efficient broilers are associated with higher oxidative stress and differential protein expression.

Variations in broiler growth and efficiency have been explained in part by differences in mitochondrial function and biochemistry in broilers. To further our knowledge in this regard, 2 experiments were carried out to determine the relationships of a) mitochondrial function and activities of various electron transport chain (ETC) complexes; b) production of H2O2, a reactive oxygen species (ROS), and its association with protein oxidation; and c) mitochondrial protein expression in liver of a single line male broilers with low or high feed efficiency (FE, n = 5 to 8 per group). Mitochondrial function and complex activities were measured polarographically and spectrophotometrically, respectively. H2O2 was measured fluorimetrically, whereas oxidized protein (carbonyls) and specific mitochondrial proteins were analyzed using Western blots. Mitochondrial function (ETC coupling) and activities of ETC complexes (I, II, III, and IV) were higher in high FE compared with low FE broilers. H2O2 and protein carbonyls were higher in the livers of low FE broilers than in high FE broilers. Whereas the expression of 4 immunoreactive proteins [NAD3 (complex I), subunit VII (complex III), cytochrome c oxidase subunits (COX) II, and COX IVb (complex IV)] were higher in low FE liver mitochondria and 2 proteins [subunit 70 (complex II) and a-ATP synthase (complex V)] were higher in high FE birds, there were no differences between groups in the expression of 18 other mitochondrial proteins. In conclusion, increases in oxidative stress in low FE broilers were caused by or may contribute to differences in mitochondrial function (ETC coupling and complex activities) or the differential expression of steady-state levels of some mitochondrial proteins in the liver. Understanding the role of oxidative stress in Low FE broilers will provide clues in understanding the cellular basis of feed efficiency.

Animal Feed↗

PubMed2004

Low feed efficient broilers within a single genetic line exhibit higher oxidative stress and protein expression in breast muscle with lower mitochondrial complex activity.

The objectives of this study were to determine the effects of low or high feed efficiency (FE) on a) protein oxidation, b) the activities of various respiratory chain complexes, and c) expression of various mitochondrial proteins in male broilers within a single genetic line. Tissue homogenate or mitochondria were isolated from breast muscle of broilers with high (0.80 +/- 0.01) and low FE (0.62 +/- 0.02). The complex activities were measured spectrophotometrically, and the levels of oxidized protein (carbonyl) and immunoreactive mitochondrial proteins were analyzed using Western blots. Protein carbonyl levels were higher in low FE compared with high FE broilers breast muscle, which indicated enhanced protein oxidation in low FE mitochondria. Activities of all respiratory chain complexes (I, II, III, IV) were higher in high FE compared with low FE broilers for breast mitochondria. Whereas the expression of immunoreactive proteins was higher in low FE muscle mitochondria for 5 mitochondrial proteins [core I, cyt c1, cyt b (complex III), COX II (cytochrome c oxidase subunit II, complex IV), and adenine nucleotide translocator (ANT1)], there were no differences between groups in the expression of 9 other respiratory chain protein subunits associated with complexex I, II, III, IV, and V. SDS-PAGE revealed a protein band of 47 kDa that was expressed at a higher level in low FE compared with high FE mitochondria. The differential expression of certain mitochondrial proteins and the 47-kDa band might be a compensatory response either to the lower complex activities or increased protein oxidation observed in low FE birds.

Animal Nutritional Physiological Phenomena↗

PubMed2004

Determination of mitochondrial function and site-specific defects in electron transport in duodenal mitochondria in broilers with low and high feed efficiency.

Duodenal mitochondria were isolated from broiler breeder males with high (0.79+/-0.01, n = 9) and low (0.63+/-0.02, n = 9) feed efficiency (FE) to assess relationships of FE with duodenal mitochondrial function and site-specific defects in electron transport. Sequential additions of adenosine diphosphate (ADP) resulted in 1) higher respiratory control ratio (RCR; an index of respiratory chain coupling) in high FE mitochondria provided succinate, and 2) higher ADP to oxygen ratio (ADP:O; an index of oxidative phosphorylation) in low FE mitochondria provided NADH-linked substrates (malate, pyruvate, or both). Basal electron leak, measured as H2O2 production, was greater in low FE mitochondria provided succinate (P = 0.08) or NADH-linked substrates. As H2O2 levels were elevated in low FE compared with high FE mitochondria by complex I (P+/-0.07) and complex II inhibition, the higher basal electron leak in low FE mitochondria was apparently due to site-specific defects in electron transport at complexes I and II. Elevations in H2O2 above basal levels indicated that high FE mitochondria may also exhibit electron transport defects at complexes I and III. Despite an ability to produce adenosine triphosphate (ATP) that was equal or superior to that demonstrated in high FE duodenal mitochondria, low FE mitochondria exhibited a greater inherent degree of electron leak. The results provide insight into the role that duodenal mitochondria play in the phenotypic expression of FE in broilers.

Adenosine Diphosphate↗

PubMed2002

Protein metabolism during an acute phase response in chickens.

Fractional rates of liver, muscle, plasma and acute phase portein synthesis were measured in chickens injected with saline or E. coli lipopolysaccharide (LPS). Male Single Comb White Leghorns were infused with a primed constant infusion of 15N-L-methionine and 2H5-L-phenylalanine into the portal vein for 2 h. Changes in plasma amino acid enrichment were similar for both amino acids reaching an apparent plateau by the 30 min sampling time. The enrichment of plasma protein-bound amino acid was measurable after 1 h of isotope infusion and increased linearly over 2h. LPS injection decreased free phenylalanine enrichment in the carotid artery (50%), and reduced tissue free methionine enrichment in the liver, pectoralis, and gastrocnemius by 16, 41, and 31% respectively. Isotopic enrichment of phenylalanine in liver protein, plasma protein and hemopexin increased in LPS injected birds relative to control birds. Fractional rates of muscle protein synthesis were not affected by LPS injection, however, liver protein, plasma protein, and hemopexin fractional synthesis rates increased 141, 161 and 266% respectively compared with untreated animals.

Acute-Phase Proteins↗

PubMed2002

Association of mitochondrial function with feed efficiency within a single genetic line of male broilers.

Studies were conducted to determine relationships between feed efficiency and mitochondrial function and biochemistry. After feed efficiency (FE; gain:feed) was determined in broiler breeder males between 6 and 7 wk of age, mitochondria were isolated from breast and leg muscle from birds with high FE (0.83+/-0.01, n = 6) and low FE (0.64+/-0.01, n = 7). Respiratory chain coupling, assessed by the respiratory control ratio (RCR), was greater in high FE breast, and leg mitochondria provided NADH-linked, but not FADH-linked, energy substrates. There were no differences, however, in the adenosine diphosphate to oxygen (ADP:O) ratio (an index of oxidative phosphorylation) when mitochondria were provided either energy substrate. Electron leak, as determined by generation of H202, was greater in the low FE than in high FE breast mitochondria. Electron leak increased following inhibition of electron transport at Complex I (with rotenone) and Complex III (with antimycin A) in low FE but not in high FE breast mitochondria. There were no differences in basal electron leak in leg mitochondria between groups, but H202 generation was elevated (P < 0.07) compared to basal values in low FE leg mitochondria after Complex I inhibition. The activities of Complexes I and II were greater in high FE breast and leg muscle mitochondria compared to those in low FE mitochondria. The results indicate that lower respiratory chain coupling in low FE muscle mitochondria may be due to lower activities of Complexes I and II and defects in electron leak and provide insight into cellular mechanisms associated with the phenotypic expression of feed efficiency in broilers.

Adenosine Diphosphate↗

PubMed2001

The hepatic extraction of plasma free amino acids and response to hepatic portal venous infusion of methionine sources in anesthetized SCWL males (Gallus domesticus).

This study was conducted to investigate the hepatic extraction of plasma free amino acids in anesthetized Single Comb White Leghorn (SCWL) males (Gallus domesticus). SCWL males were anesthetized and implanted with cannulae in the carotid artery, hepatic vein, hepatic portal vein and the left hepatic duct. Free amino acids in plasma and bile were determined before, during and after 30-min infusions of Saline (control), DL-Methionine (DL-Met) or DL-2-hydroxy-4-methylthio-butanoic acid (DL-HMB) into the hepatic portal vein. Hepatic extraction rates (HER) of amino acids were calculated based on the concentration of amino acids in plasma multiplied by estimations of blood flow in the hepatic portal vein, hepatic artery and hepatic vein. For the non-essential amino acids, alanine had the highest HER (46%). The liver also removed more than 20% of hepatic inflow of tyrosine and asparagine with substantial extraction (14-18%) of serine, glycine and glutamine, also. In contrast, less than 5% of hepatic inflow of glutamate and cystine were removed by liver. For the essential amino acids, HER for methionine, histidine and phenylalanine were 30, 14 and 17%, respectively, with less than 5% for branched-chain amino acids, lysine, arginine and threonine. Biliary secretion of amino acids represented a small percentage (<0.2%) of total hepatic extraction turnover of the amino acids. Infusion of methionine sources, DL-Met and DL-HMB, had no effect on hepatic metabolism of amino acids other than methionine. The results demonstrated for the first time, the hepatic extraction of circulating free amino acids in avian species in vivo.

Amino Acids↗

PubMed1997

Effect of Santoquin and oxidized fat on liver and intestinal glutathione in broilers.

Experiments were conducted to determine effects of Santoquin (ethoxyquin) and oxidized fat on liver and intestinal reduced (GSH) and oxidized (GSSG) glutathione, and pulmonary hypertension syndrome (PHS) mortality. Male broilers were randomly assigned in a 2 x 2 factorial consisting of 3.5% normal (NF) or oxidized (OxF) fat with or without ethoxyquin (E). Body weights and feed intake were monitored weekly, and tissues obtained at 3 and 7 wk for GSH and GSSG analysis. Compared to the NF group, NF/E gained more weight during the starter (0 to 3 wk), but not the grower (4 to 7 wk) period. Birds fed NF/E or NF exhibited greater feed efficiency in the starter period and greater gains during the starter and grower periods than birds fed OxF or OxF/E. No differences in PHS mortality between treatments were observed. Birds fed OxF exhibited lower liver GSSG at 3 wk than the other groups, but there were no differences in liver GSH. Duodenal GSH was higher in birds fed OxF/E than in birds of NF group at 3 and 7 wk. Ileal GSH was higher at 3 wk in OxF/E birds than in OxF birds, but no differences were observed at 7 wk. All tissues exhibited higher GSH levels at 7 wk than at 3 wk. Birds fed ethoxyquin, regardless of fat source, exhibited higher duodenal GSH at 3 and 7 wk and higher ileal GSH at 3 wk than birds that did not receive ethoxyquin. Higher GSH would be beneficial by enhancing protection of intestinal cells to deleterious effects of toxins or other forms of oxidative stress.

Animals↗

PubMed1995

Effect of alpha-tocopherol on antioxidants, lipid peroxidation, and the incidence of pulmonary hypertension syndrome (ascites) in broilers.

Research has demonstrated a compromised antioxidant capacity in broilers with pulmonary hypertension syndrome (PHS). Thus, the objective of the present study was to assess the effects of vitamin E on PHS-induced mortality, tissue antioxidants, and plasma lipid peroxides in male broilers. Control broilers were provided normal ventilation but others, maintained under low ventilation conditions to induce PHS, were randomly assigned to nonimplanted (NI), placebo (PL), or vitamin E (VE) implanted groups. The VE implant released a total of 15 mg of alpha-tocopherol from 0 to 3 wk of age. Tissues and blood samples were obtained at 3 and 5 wk of age from birds with (PHS+) and without (PHS-) PHS. Five-week PHS cumulative mortality was lowered by alpha-tocopherol with mortality rates of 3.6, 4.2, 11.9, and 11.8%, for Controls, VE, NI, and PL groups, respectively. The PHS+ birds exhibited lower body weights, higher hematocrit, right ventricular hypertrophy, lower alpha-tocopherol and glutathione (GSH) concentrations in liver and lung, as well as indicators of oxidative stress, including elevated plasma lipid peroxides and lower oxidized GSH in liver and erythrocytes, at 5 wk of age. All birds exhibited lower erythrocyte catalase activity at 5 than at 3 wk of age. An improved antioxidant capacity was observed in VE birds, including higher liver and lung alpha-tocopherol at 3 and 5 wk, higher liver GSH at 3 wk, and lower plasma lipid peroxide values at 5 wk of age. Direct correlations observed between body weight and plasma lipid peroxides at 3 wk (r = .45) and between right ventricular hypertrophy and plasma lipid peroxides at 5 wk (r = .48), suggests that lipid peroxidation plays a role in the etiology of PHS. The results indicate that the VE implant was effective in lowering PHS-induced mortality in broilers apparently by attenuating processes leading to lipid peroxidation.

Analysis of Variance↗

PubMed1995

Pulmonary arteriole hypertrophy in broilers with pulmonary hypertension syndrome (ascites).

Two experiments were conducted to determine the effect of low ventilation or cool temperature environments on pulmonary arteriole hypertrophy. Male broilers were maintained under control or low ventilation conditions in Experiment 1, whereas male broiler breeder by-product chicks were exposed to cool temperature conditions in Experiment 2. Birds were randomly selected for histological evaluation of lung tissue in both experiments. In Experiment 1, birds that had pulmonary hypertension syndrome (PHS+) exhibited a greater degree of inflammation of lung tissue at 5 and 7 wk of age than controls or birds that did not have PHS (PHS-). These PHS+ birds also had higher numbers of cartilaginous osseous nodules at 3 and 7 wk of age than controls. Morphometric analyses revealed that PHS+ birds in Experiment 1 had a thicker medial layer associated with 100 to 200 microns diameter pulmonary arterioles at 7 wk of age, and 50 to 100 microns arterioles at 3 and 7 wk of age than PHS- or control birds. In Experiment 2, PHS+ birds exhibited a thicker medial layer in pulmonary arterioles at 7 wk of age than did PHS- birds, but there were no differences in medial layer thickness at 5 wk of age nor were there differences in the degree of inflammation or amount of osseous nodule formation between PHS+ and PHS- birds at 5 and 7 wk of age. Thus, pulmonary arteriole hypertrophy was observed in birds having PHS in response to both low ventilation and cool temperature environments and this hypertrophy occurred with or without a coincident inflammatory response in lung tissue.

Animals↗

PubMed1993

Compromised antioxidant status associated with ascites in broilers.

Tissue infiltration by white blood cells in poultry with ascites has been reported, which could alter endogenous antioxidant status from oxidant stress. Therefore, the objective of this study was to determine the effect of ascites on ascorbic acid, tocopherol, and glutathione (GSH), which are major intracellular antioxidants. Broiler chicks (1 day) were placed in environmental chambers maintained with normal ventilation (Control) or under low ventilation, which produced a cumulative mortality of 31.3% due to ascites. At 3, 5, and 7 wk, lung and liver tissues were obtained from control birds (n = 5) and from birds in the low ventilation chamber with (ASC, n = 5) or without (NASC, n = 5) overt symptoms of ascites. There were no differences in tissue ascorbate, tocopherol, and GSH between the control and NASC groups with the exception of hepatic tocopherol at 7 wk, which was higher (P < .05) in NASC birds than in ASC and control birds. In general, lung and liver concentrations of all three antioxidants were lower (P < .05) in ASC birds than in NASC and control groups. Uric acid, a product of purine metabolism, was lower (P < .05) in liver and lung in ASC birds at 3 wk compared with control birds. However, uric acid concentrations in ASC birds were higher (P < .05) in the lung and serum at 5 wk, and in the liver at 7 wk compared with NASC and control birds.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Effect of diethyl maleate on glutathione, hepatic and renal cortical perfusion, and portal 6-ketoPGF1 alpha and TxB2 levels in swine.

1. Effects of diethyl maleate (DEM) mediated glutathione (GSH) depletion on hepatic and renal cortical blood flow (perfusion), plasma GSH, and portal prostacyclin (6-ketoPGF1 alpha) and thromboxane (TxB2) were determined in anaesthetized swine. 2. Although DEM depleted hepatic GSH to 25% of control, plasma GSH increased 10-fold in comparison to controls. DEM caused a drop in blood pressure and renal cortical perfusion but had no effect on hepatic perfusion or portal 6-ketoPGF1 alpha or TxB2 levels. 3. Possibly, the unexpected rise in plasma GSH may have inhibited prostanoid synthesis, preventing any alterations in tissue perfusion that may have occurred following tissue GSH depletion.

6-Ketoprostaglandin F1 alpha↗

PubMed1991

Indomethacin attenuation of hepatic perfusion and plasma 6-ketoPGF1 alpha elevations following glutathione depletion in rabbits.

Glutathione (GSH) is important in detoxification and regulating cyclooxygenase activity. Since the liver has high levels of GSH, xenobiotic-induced changes in hepatic GSH could affect hepatic tissue blood perfusion (HP) via alterations in prostaglandin synthesis. In anesthetized male New Zealand rabbits, elevating GSH with GSH monoethyl ester had no affect on HP. Treatment of rabbits with diethyl maleate to deplete GSH also had no affect on HP in animals previously given GSH monoethyl ester. However, HP increased within 20 min in rabbits treated with diethyl maleate prior to GSH monoethyl ester. In another experiment, a similar rise in HP following GSH depletion was accompanied by arterial plasma 6-ketoPGF1 alpha (the stable metabolite of prostacyclin) levels that were 4-times higher than in the controls. Plasma TxB2 (the stable metabolite of thromboxane) also increased following diethyl maleate, but only to levels that were 25-times lower than for 6-ketoPGF1 alpha. Since indomethacin blocked the rise in HP, as well as the increases in 6-ketoPGF1 alpha and TxB2, these results indicate changes in HP may occur following GSH depletion as a result of increased synthesis of one or more arachidonic acid metabolites and implicate prostacyclin as a possible mediator of this phenomenon.

6-Ketoprostaglandin F1 alpha↗

PubMed1990

Indomethacin attenuation of celiac blood flow hyperemia following glutathione depletion.

The effect of glutathione (GSH) depletion on mean celiac blood flow (MCBF) was determined in domestic fowl. Diethyl maleate (DEM, 1 mL/kg body wt) decreased hepatic and duodenal GSH to approximately 15% of control. This GSH depletion was associated with an increase in MCBF and decreases in mean arterial blood pressure (MABP) and celiac vascular resistance (CVR). While indomethacin attenuated the rise in MCBF, this cyclooxygenase inhibitor had no effect on the decrease of MABP or CVR which occurred following DEM treatment. The results indicate that GSH depletion may increase vasodilatory prostaglandin synthesis since elevations in MCBF were attenuated by cyclooxygenase inhibition.

Animals↗