Sir2-dependent activation of acetyl-CoA synthetase by deacetylation of active lysine

Acetyl-coenzyme A (CoA) synthetase (Acs) is an enzyme central to metabolism in prokaryotes and eukaryotes. Acs synthesizes acetyl CoA from acetate, adenosine triphosphate, and CoA through an acetyl-adenosine monophosphate (AMP) intermediate. Immunoblotting and mass spectrometry analysis showed that Salmonella enterica Acs enzyme activity is posttranslationally regulated by acetylation of lysine-609. Acetylation blocks synthesis of the adenylate intermediate but does not affect the thioester-forming activity of the enzyme. Activation of the acetylated enzyme requires the nicotinamide adenine dinucleotide-dependent protein deacetylase activity of the CobB Sir2 protein from S. enterica. We propose that acetylation modulates the activity of all the AMP-forming family of enzymes, including nonribosomal peptide synthetases, luciferase, and aryl- and acyl-CoA synthetases. These findings extend our knowledge of the roles of Sir2 proteins in gene silencing, chromosome stability, and cell aging and imply that lysine acetylation is a common regulatory mechanism in eukaryotes and prokaryotes.     

Evaluation of the bispectral index as an indicator of degree of central nervous system depression in isoflurane-anesthetized horses

OBJECTIVE: To determine whether the bispectral index (BIS) can be used as an indicator of degree of CNS depression in isoflurane-anesthetized horses. ANIMALS: 10 Standardbred and 6 Norwegian cold-blooded trotter stallions admitted for routine castration. PROCEDURE: A 2-channel referential electrode configuration was used to record EEG for calculation of BIS by the EEG monitor. The BIS was calculated before (awake) and after (sedated) administration of detomidine (0.01 mg/kg of body weight, IV) and butorphanol (0.01 mg/kg, IV). Anesthesia was induced with ketamine hydrochloride (2.5 mg/kg, IV) and diazepam (0.04 mg/kg, IV) and maintained with isoflurane delivered in oxygen. The BIS was calculated after 30 minutes of equilibration at an end-tidal isoflurane concentration of 1.4% (n = 8) or 1.9% (8) and recorded continuously during surgery. RESULTS: Bispectral index was significantly less in sedated and anesthetized horses, compared with awake horses. However, BIS was not significantly different between sedated and anesthetized horses. Mean BIS in horses anesthetized at 1.9% isoflurane was significantly greater, compared with horses anesthetized at an end-tidal concentration of 1.4%. Four horses in the 1.4% group moved during surgery, and BIS increased immediately prior to movement in 2 of these horses. CONCLUSIONS AND CLINICAL RELEVANCE: BIS is not a precise indicator of degree of CNS depression in isoflurane-anesthetized horses. Thus, determination of BIS may not be a useful technique for monitoring anesthetic depth in isoflurane-anesthetized horses.     

Interactions between dietary crude protein and essential amino acid intake on performance in broilers

1. The effect of diets with increasing concentrations of crude protein at either fixed essential amino acid concentrations or at fixed essential amino acid:dietary crude protein ratios on performance was examined in 1- to 4-week-old male Cobb chicks. Increasing crude protein intakes at constant essential amino acid concentrations was carried out at two dietary energy contents. 2. Increasing crude protein resulted in a linear decrease in feed intake while weight gain and feed efficiency changed quadratically with a smaller positive effect at the highest crude protein intakes. Feed intake decreased and feed efficiency increased with higher dietary energy and interactions between protein and energy were significant. Abdominal fat content and the efficiency of protein retention decreased with increasing dietary protein intake. 3. Using constant essential amino acid:crude protein ratios at increasing crude protein intakes resulted in (Trial 3) feed intake, weight gain and feed efficiency all increasing before reaching a plateau. Abdominal fat decreased with protein intake and the efficiency of protein retention was quadratic, decreasing at the higher protein intakes. 4. Multiple regression analysis of the results of the three trials indicated that partition of energy intake into maintenance, fat-free tissue growth, fat and the energy required to transform protein intake in excess of retention explained more than 98% of variation. 5. It is proposed that broiler performance at the lower protein intakes was limited by either non-essential amino acid (Trials 1 and 2) or essential amino acid (Trial 3) intake whereas at high protein intakes the decreased efficiency of amino acid utilisation after growth requirements are fulfilled resulted in poorer performance.     

Genetic resistance to Salmonella typhimurium in two lines of chickens selected as resistant and sensitive on the basis of heterophil/lymphocyte ratio

1. A study was conducted to test for the validity of the heterophil/lymphocyte (H/L) ratio as a criterion for selection for resistance to Salmonella typhimurium in chickens. 2. An infective dose of S. typhimurium was given, directly in the crop, to two groups of chicken selected as Resistant (R) and Sensitive (S) on the bases of H/L ratio. The 99% lower confidence limit was used as a borderline; individuals below the limit were considered R and those above S. Many aspects of immune response were compared, namely: H/L ratio, antibody titre, cellular immunity, phagocytic activity, cortisol concentration, bursa and body weight. 3. The R group exceeded the S in all the studied variables of the immune response, indicating the possibility of using the H/L ratio and its confidence limit to select for general resistance. 4. Due to the within-strain variability in resistance, it seems that immunological and genetic studies should take into consideration separation of individuals into R and S before grouping. Failing to do so might lead to erroneous conclusions as a difference may simply be due to the different numbers of R and S included in each group.     

Construction and immunogenicity studies of recombinant fowl poxvirus containing the S1 gene of Massachusetts 41 strain of infectious bronchitis virus

The spike 1 (S1) surface glycoprotein of infectious bronchitis virus (IBV) is the major inducer of the generation of virus neutralizing antibodies, and the administration of purified S1 has been shown to elicit a protective immune response against virulent virus challenge. On the basis of these observations, recombinant fowl poxvirus (rFPV) containing a cDNA copy of the S1 gene of IBV Mass 41 (rFPV-S1) was constructed and its immunogenicity and vaccine potential were evaluated. Initially, rFPV-S1 was shown to express the S1 in vito by indirect immunofluorescence staining and western blot analyses. Later, in vivo expression was demonstrated by the detection of IBV-specific serum immunoglobulin G and neutralization antibodies in the sera of chickens immunized with rFPV-S1. That the recombinant virus elicited anti-IBV protective immunity was indicated by the manifested, relatively mild clinical signs of disease, decreased titers of recovered challenge virus, and less severe histologic changes of the tracheas in virulent IBV Mass 41-challenged chickens previously receiving rFPV-S1 as compared with parental fowl poxvirus (FPV)-vaccinated control birds. In contrast, chickens immunized with either recombinant or parental FPV were resistant to a subsequent virulent FPV challenge. As to a preferred method of immunization, wing web administration appeared to be superior to the subcutaneous route because a greater percentage of birds vaccinated by the former protocol exhibited an anti-IBV humoral immune response. Thus, rFPV-S1 has potential as a poultry vaccine against both fowl pox and infectious bronchitis.     

Effects of restricted feeding on physiological stress parameters in growing broiler breeders

1. In previous studies, a lack of agreement in measurements of plasma corticosterone concentrations and heterophil:lymphocyte (H/L) ratio as physiological indices of stress, caused by hunger and frustration in restricted-fed broiler breeders, was observed. It could be suggested that the differences between previous studies were caused by differences in duration of restriction and time of the day of the measurements. Therefore, in the present study the plasma corticosterone concentration and the H/L ratio were again determined in restricted- and ad libitum-fed growing broiler breeders, taking possible causes of disagreement between previous studies into account. In addition, we measured the daily rhythm in body temperature and heart rate, and the corticosterone responses to an acute stressor as physiological indices of stress. 2. Female broiler breeders (64 per treatment, housed in groups of 4 birds) were used in the experiment. Behaviour, baseline plasma corticosterone concentrations and H/L ratio were determined at 21 d of age (immediately after the start of food restriction), and at 42 and 63 d of age. Body temperature, heart rate and activity were measured by radiotelemetry for 36 h at 49 and 70 d of age. In addition, the plasma corticosterone response to acute stress (5 min manual restraint) was measured at 77 or 78 d of age. 3. Restricted broiler breeders had higher plasma corticosterone concentrations at 42 and 63 d of age, but no differences in H/L. ratio were found between restricted birds and unrestricted control birds. Restricted broiler breeders had a higher corticosterone response to 5 min manual restraint than unrestricted birds. Restricted birds displayed a clear day-night rhythm in body temperature, heart rate and activity whereas such a rhythm was blunted in ad libitum-fed birds. 4. It is discussed that some physiological differences (plasma corticosterone concentrations, body temperature and heart rate) between ad libitum-fed and restricted broiler breeders may have been caused by differences in metabolic rate as well as by differences in the level of stress. It is concluded that a combination of behavioural measurements and a wide range of physiological parameters should be used for the assessment of stress in growing broiler breeders.     

Seasonal interaction of serum vitamin D concentration and bone density in alpacas

OBJECTIVE: To evaluate temporal changes in bone mineral density associated with seasonal variation in serum vitamin D, calcium, and phosphorus concentrations in alpacas. ANIMALS: 5 healthy mature neutered male alpacas. PROCEDURE: Metacarpal bone mineral density was measured at 4 times during a year. Each time alpacas were weighed, blood was collected for determination of serum calcium, phosphorus, and vitamin D concentrations, and samples of feed were analyzed for nutrient content. Vitamin D status was determined by use of an assay that measured serum 25-hydroxycalciferol concentration. Effects of changes in serum vitamin D, calcium, and phosphorus concentration and body weight with season on bone mineral density were determined. RESULTS: Bone mineral density, body weight, and serum vitamin D and phosphorus concentrations varied with season. Bone mineral density, serum vitamin D concentration, and body weight also varied among individual alpacas. Serum vitamin D concentration was lower in January than the previous October and increased from May to the following September. The decrease in bone mineral density lagged behind the decrease in serum vitamin D concentration and was lower in May, compared with the previous October. Body weight was lower in May than the previous October or following September. Solar radiation was highest in July and lowest in December. CONCLUSIONS AND CLINICAL RELEVANCE: Seasonal changes in bone mineral density are associated with changes in serum vitamin D concentrations in alpacas. Changes in bone mineral density associated with a decline in serum vitamin D concentration may predispose some alpacas to developing fractures minimal trauma.     

Relationship between incidence of gastric dilatation-volvulus and biometeorologic events in a population of military working dogs

OBJECTIVE: To calculate the monthly incidence of gastric dilatation-volvulus (GDV) in a population of military working dogs during a 5-year period and determine whether there was an association with synoptic climatologic indices. SAMPLE POPULATION: Medical records of all military working dogs housed at Lackland Air Force Base,Tex, from Jan 1, 1993 to Dec 31, 1997. PROCEDURE: Confirmed cases of GDV were identified from evaluation of medical records and used to calculate incidence of GDV. Factor analysis of local climatologic data was used to classify each day into 1 of 8 meteorologically homogeneous types of days for this location. Occurrence of GDV was compared with frequency of occurrence of synoptic climatologic days. RESULTS: 48 cases of GDV were identified from January 1993 through December 1997 Mean monthly incidence was 2.5 cases/1,000 dogs at risk (range, 0 to 18.5 cases/1,000 dogs; median, 2.5 cases/1,000 dogs). A seasonal increase in incidence of GDV was detected, because half of the episodes were during November, December, and January. An association with a specific synoptic climatologic day was not detected. CONCLUSIONS AND CLINICAL RELEVANCE: Seasonal fluctuations in incidence of GDV may be associated with external factors that precipitate physiologic changes resulting in GDV. Although a specific cause-effect relationship was not documented, clinicians must be alert for the potential of seasonal variation in incidence of GDV and accordingly heighten their index of suspicion for the condition, particularly in populations of dogs that are predisposed to development of GDV.     

Effect of tilmicosin on chemotactic, phagocytic, and bactericidal activities of bovine and porcine alveolar macrophages

OBJECTIVE: To evaluate chemotactic, phagocytic, and bactericidal activities of bovine and porcine alveolar macrophages (AM) exposed to tilmicosin. ANIMALS: 12 healthy calves and 12 healthy pigs. PROCEDURES: Lungs were obtained immediately after euthanasia; AM were collected by means of bronchoalveolar lavage and density gradient centrifugation. Chemotactic activity was evaluated by exposing AM to lipopolysaccharide or macrophage inhibitory peptide during incubation with tilmicosin. Phagocytic activity was evaluated by incubating AM with tilmicosin for 24 hours and then with tilmicosin-resistant Salmonella serotype Typhimurium. Bactericidal activity was evaluated by incubating AM with tilmicosin (0, 10, or 20 microg/ml for bovine AM; 0 or 10 microg/ml or 10 microg/ml but washed free of tilmicosin for porcine AM) and then with Mannheimia haemolytica (bovine AM) or with Actinobacillus pleuropneumoniae or Pasteurella multocida (porcine AM). RESULTS: Tilmicosin had no significant effects on chemotactic or phagocytic activities of bovine or porcine AM. The time-course of bactericidal activity was best described by polynomial equations. Time to cessation of bacterial growth and area under the time versus bacterial number curve were significantly affected by incubation of AM with tilmicosin. CONCLUSIONS AND CLINICAL RELEVANCE: Results show that bactericidal activity of bovine and porcine AM was enhanced by tilmicosin, but not in proportion to the reported ability of AM to concentrate tilmicosin intracellularly. With or without exposure to tilmicosin, the time-course of bactericidal activity of bovine AM against M haemolytica and of porcine AM against A pleuropneumoniae or P multocida was too complex to be reduced to a simple linear equation.