Preliminary Studies on the Concentration of Na+,K+-ATPase in Skeletal Muscle of Draught Cattle in Mozambique: Effect of Sex,Age and Training

The effect of training on the potential for work in draught cattle was assessed by measuring the Na⁺,K⁺-ATPase in the muscle cell membrane and the elevation in the concentration of K⁺ in plasma during exercise. Biopsies of the semitendinosus muscle and venous blood samples were taken from the cattle used for draught work in Mozambique. No differences were found in the plasma ion or Na⁺,K⁺-ATPase concentrations in samples taken from Nguni, Africander and Angoni breeds. There were no significant differences in plasma ions (Na⁺, K⁺ and Cl^–) or muscle Na⁺,K⁺-ATPase concentrations between the Angoni males and females, although the males showed an increase in Na⁺,K⁺-ATPase with age, while the females showed a decrease. The increase in males might be attributed to their higher level of activity in the herds than that of females. After a training period of 15 days, a significant increase in Na⁺,K⁺-ATPase concentration in semitendinosus muscle was found in Angoni cattle. In females, this was significant after 8 days of training (about 30%); in males after 15 days of training (about 16%). On day 15, there was a reduction in the elevation of plasma K⁺ during the 2 h of draught work, indicating an increased capacity of the Na⁺,K⁺ pumps to maintain the extracellular K⁺ concentration in working muscles and a possible delay in the moment of fatigue.     

Insulin accelerates recovery from QRS complex widening in a frog heart model of hyperkalemia

Hyperkalemia is one of the most common electrolyte disorders. By injecting various concentrations of potassium chloride (KCl) solutions intravenously into bullfrogs, we demonstrated characteristic electrocardiogram (ECG) abnormalities of hyperkalemia in frog hearts. The widened QRS complexes induced by 100 mM KCl injection were accompanied by an increase in the resting membrane potential in cardiomyocytes and a decreased slope of phase 0 in the action potential. Recording both ECG waveforms and the cardiac action potential enabled us to reveal the mechanisms of hyperkalemia-induced ECG abnormalities. Additionally, pre-treatment with insulin, a powerful stimulator of Na⁺/K⁺-ATPase activity, significantly accelerated the recovery from the widened QRS complexes in the ECG, demonstrating a pronounced shift of extracellular potassium ions into the intracellular space.     

A single injection of periostin decreases cardiac voltage-gated Na+ channel in rat ventricles

Changes in electrophysiological properties, such as ion channel expression and activity, are closely related to arrhythmogenesis during heart failure (HF). However, a causative factor for the electrical remodeling in HF has not been determined. Periostin (POSTN), a matricellular protein, is increased in heart tissues of patients with HF. In the present study, we investigated whether a single injection of POSTN affects the electrophysiological properties in rat ventricles. After male Wistar rats were intravenously injected with recombinant rat POSTN (64 µg/kg, 24 hr), electrocardiogram (ECG) was recorded. Whole-cell patch clamp was performed to measure action potential (AP) and Na⁺ current (I_Na) in isolated ventricular myocytes. Protein expression of cardiac voltage-gated Na⁺ channel (Na_V1.5) in isolated ventricles was examined by Western blotting. In ECG, POSTN-injection significantly increased RS height. POSTN-injection significantly delayed time to peak in AP and decreased I_Na in the isolated ventricular myocytes. POSTN-injection decreased Na_V1.5 expression in the isolated ventricles. It was confirmed that POSTN (1 µg/ml, 24 hr) decreased I_Na and Na_V1.5 protein expression in neonatal rat ventricular myocytes. This study for the first time demonstrated that a single injection of POSTN in rats decreased I_Na by suppressing Na_V1.5 expression in the ventricular myocytes, which was accompanied by a prolongation of time to peak in AP and an increase of RS height in ECG.     

Effect of Cadmium in Feed on Organs and Meat Colour of Growing Pigs

One hundred and ninety-two barrows (Duroc × Landrace × Yorkshire, initial weight 27.7 kg) were used to investigate the effects of cadmium in feed on the function of selected organs and meat colour of growing pigs. The pigs were randomly allocatted into four different treatments. Each treatment included three replications with 16 pigs per replicate. The animals were fed corn–soybean basal diet and supplemented with 0, 0.5, 5.0, 10.0 mg/kg cadmium (as CdCl₂), respectively. The feeding trial ended when the average body weight of the pigs reach 90 kg. The results showed that, compared with controls, addition of 10 mg/kg cadmium to the diet resulted in significant elevations of relative weight of liver and spleen by 18.3% (p < 0.05) and 19.7% (p < 0.05) respectively, and of serum glutamic-pyruvic transaminase (GPT) and glutamic-oxaloacetic transaminase (GOT) activities by 17.8% (p < 0.05) and 27.4% (p < 0.05) respectively; and significant decreases of Na⁺/K⁺-ATPase activity in the liver by 24.6% (p < 0.05), the redness of longissimus dorsi by 26.6% (p < 0.05) and 24.9% (p < 0.05) at 0.75 h and 16 h post mortem, respectively, and of the myoglobin content of longissimus dorsi by 19.4% (p < 0.05). No changes were found in these indices above when the pigs were fed the diet supplied with 0.5 or 5 mg/kg cadmium (p > 0.05), nor in renal functions among cadmium-treatment treatments (p > 0.05) as indicated is the activities of urinary N-acetyl-β-d-glucosaminidase (NAG) and alkaline phosphatase (ALP) and the content of urinary protein. The study indicated the adverse effects of 10 mg/kg cadmium in feed on liver functions and meat colour of growing pigs.     

Factors affecting disease manifestation of toxocarosis in humans: Genetics and environment

Toxocara canis is regarded as the main cause of human toxocarosis but the relative contribution of T. cati is probably underestimated; serological and other diagnostic methods used in most studies of this zoonotic disease do not distinguish between the two parasites. The definitive hosts for T. canis are caniidae. Pups generally have higher infection rates than adult animals and are a major source of eggs in the environment. Humans usually acquire T. canis infection by accidental ingestion of embryonated eggs or encapsulated larvae from the environment or contaminated food, such infections may lead to visceral larva migrans (VLM), ocular larva migrans (OLM) or covert toxocarosis (CT). Although a mixed Th1- and Th2-mediated immunological response, particularly with high levels of IgE and eosinophilia is observed, the underlying mechanisms of molecular and immunopathogenesis for the development of the symptomatic syndromes of VLM, OLM, or of asymptomatic CT are largely unclear. Studies have indicated that immunological defences against various infectious diseases may be highly influenced by complex interactions of environmental and host genetic factors e.g. MHC class I and II, also known as human leucocyte antigen (HLA). Toxocara spp. infections are associated with a polarized CD4⁺ Th2 response with high IgE levels and eosinophilia, mediated mainly by HLA class II molecules. Associations have been made between HLA class II and pathological severity and host genetic effects on exposure to infection. Recent research suggests Foxp3⁺ CD4⁺CD25⁺-expressing T regulatory (Treg) cells play a role in regulation of the immunopathology of granulomas in experimental toxocaral granulomatous hepatitis and in enhanced expression of TGF-β1, which is an important factor for the local survival and function of Treg observed during T. canis invasion in the mouse small intestine, liver, muscle, and brain. Since the potential susceptibility loci HLA class II molecules, are considered involved in the regulation of a Th2-dominant immunity which is highly controlled by Foxp3⁺ CD4⁺CD25⁺ Treg cells by stimulation through TGF-β1, which thus provides a beneficial environment to T. canis larvae but severe injuries to local organs. However, TGF-β1 variant Leu10Pro known to be involved in disease severity warrants further elucidation as this too may have a role in the severity of human toxocarosis. Exploration of TGF-β1 polymorphism, Foxp3⁺ CD4⁺CD25⁺ Treg cells, and MHC polymorphisms may allow insight into the contribution made by environmental and genetic factors in influencing disease syndrome type and severity in humans with toxocarosis.     

Effect of Tributyltin,Cadmium, and Their Combination on Physiological Responses in Juvenile Grass Carp

Tributyltin (TBT) and cadmium (Cd) are two common pollutants in aquatic environments. This study was designed to examine the physiological responses of juvenile Grass Carp Ctenopharyngodon idella to TBT, Cd, and their combination. Fish were apportioned into a control group, a TBT group (7.5 μg/L), a Cd group (2.97 mg/L), and a TBT–Cd group (7.5 μg/L TBT, 2.97 mg/L Cd²⁺) for 7 d. The following activities were measured: Na⁺,K⁺-ATPase in gill tissues; nitric oxide synthase (NOS), acetylcholinesterase (AChE), and monoamine oxidase (MAO) in brain tissues; and lipid peroxidation (LPO), malondialdehyde (MDA), total antioxidative capacity (T-AOC), and glutathione (GSH) in liver tissues. Cadmium-induced stress was suggested by alterations in antioxidant responses (MDA, LPO, and T-AOC) and neurological parameters (AChE, MAO, and NOS). Cadmium also induced Na⁺,K⁺-ATPase and GSH activity. Compared with the responses among the Cd group, the combination of TBT and Cd not only decreased the level of GSH and Na⁺,K⁺-ATPase but also increased the levels of MDA, LPO, AChE, MAO, and NOS. These results suggest that a combination of TBT and Cd could reduce the adverse effects of Cd on Grass Carp. However, the exact mechanisms for the combined effects TBT and Cd on these biomarkers require further investigation.

Received September 28, 2015; accepted April 17, 2016 Published online August 2, 2016     

Vaccination of calves with Mycobacteria bovis Bacilli Calmete Guerin (BCG) induced rapid increase in the proportion of peripheral blood γδ T cells

Changes in the proportion of peripheral blood T cell subsets after subcutaneous inoculation of cattle with Mycobacterium bovis Bacille Calmette-Guerin (BCG) were studied. Calves were injected with approximately 8 × 10⁶ BCG bacillus and blood samples collected at weekly intervals for flow-cytometric analyses to determine the proportion of CD4⁺, CD8⁺ and γδ T cells. In addition, whole blood samples were stimulated in vitro with M. bovis purified protein derivative (PPD) and the secreted IFN-γ quantified by ELISA. Results showed cellular and cytokine changes which could be categorized into three phases. The first phase occurred within the first 2 weeks after vaccination involving an increase in proportion of WC1⁺ γδ T cells and a concomitant increase in the secretion of IFN-γ. These two responses peaked at 2 weeks and waned thereafter. The second phase involved an increase in the CD4/CD8 ratio as a result of an increase in the proportion of CD4⁺ T cells between 4 and 6 weeks. The third phase involved a decrease in the CD4/CD8 ratio due to an increase in the proportion of CD8⁺ T cells between 8 and 10 weeks. Surprisingly, the IFN-γ response was associated with changes in the γδ rather than the CD4⁺ or CD8⁺ T cells, suggesting that this cytokine was secreted by γδ-T cells. These results are consistent with the reported ability of γδ T cells to act rapidly and bridging the innate and classically adaptive immune responses.     

Inhibitory effects of SKF96365 on the activities of K+ channels in mouse small intestinal smooth muscle cells

In order to investigate the effects of {"type":"entrez-protein","attrs":{"text":"SKF96365","term_id":"1156357400","term_text":"SKF96365"}}SKF96365 (SKF), which is a non-selective cationic channel blocker, on K⁺ channel currents, we recorded currents through ATP sensitive K⁺ (I_KATP), voltage-gated K⁺ (I_Kv) and Ca²⁺ activated K⁺ channels (I_BK) in the absence and presence of SKF in single small intestinal myocytes of mice with patch-clamp techniques. SKF (10 µM) reversibly abolished I_KATP that was induced by cromakalim (10 µM), which is a selective ATP sensitive K⁺ channel opener. These inhibitory effects were induced in a concentration-dependent and voltage-independent manner. The 50% inhibitory concentration (IC₅₀) was 0.85 µM, which was obviously lower than that reported for the muscarinic cationic current. In addition, SKF (1 µM ≈ the IC₅₀ value in I_KATP suppression) reversibly inhibited the I_Kv that was induced by repetitive depolarizing pulses from −80 to 20 mV. However, the extent of the inhibitory effects was only ~30%. In contrast, SKF (1 µM) had no significant effects on spontaneous transient I_BK and caffeine-induced I_BK. These results indicated that SKF inhibited ATP sensitive K⁺ channels and voltage-gated K⁺ channels, with the ATP sensitive K⁺ channels being more sensitive than the voltage-gated K⁺ channels. These inhibitory effects on K⁺ channels should be considered when SKF is used as a cationic channel blocker.     

Studies on the Mechanism of Diarrhoea Induced by Escherichia coli Heat-Stable Enterotoxin (STa) in Newborn Calves

Enterotoxigenic Escherichia coli (ETEC) produces a heat-stable enterotoxin (STa) that binds to and activates a putative intestinal receptor, guanylate cyclase, causing an increase in the intracellular levels of cyclic guanosine monophosphate (cGMP). Using flow cytometry and ¹²⁵I-STa binding assays, we studied the distribution of STa-receptors on enterocytes isolated from different segments of the newborn calf's intestinal tract. We also investigated the effect of STa on the intracellular levels of cGMP and ion transport to the intestinal lumen. More STa-receptors were found on enterocytes prepared from the ileum than on enterocytes obtained from the other segments of the intestinal tract. Guanylate cyclase activity was higher in the ileum of STa-challenged calves than in the ileum of control calves. No changes were observed in the guanylate cyclase activity of the other intestinal segments of the STa-challenged and control calves. Na⁺ levels, as measured by atomic absorption spectroscopy, were significantly increased in the luminal contents of the ileum of STa-challenged calves, whereas serum Cl^– levels were significantly lower in the STa-challenged calves than in control calves. This study supports previous observations on the role of guanylate cyclase in the initiation of STa-induced secretory diarrhoea and suggests that Na⁺/Cl^– coupling may be the major mechanism for the loss of ions in the diarrhoeal response that is mostly induced in the ileum of newborn calves.     

The γδ cells as marker of non-seroconverted cattle naturally infected with Mycobacterium avium subspecies paratuberculosis

Early diagnosis of MAP infection is a pressing need to enable efficient intervention with the spread of MAP infection in herds. Hence, study of lymphocyte subsets and their expressed adhesion molecules could contribute in defining a distinct diagnostic marker (or markers) at the subclinical period of the infection that could in turn facilitate the development of effective diagnostic approach. In accordance with this objective, milk and blood samples were collected from two groups of cattle naturally infected with MAP and their corresponding negative controls. Group (C) comprised 3-4 year-old ELISA negative/PCR positive-cattle that were considered as subclinical seronegative low shedder group (early stage). Group (A) included 6-8 year-old ELISA positive-cattle, which were considered as a clinical seropositive group (late stage). Flow cytometry of B cells, CD8⁺, CD4⁺ and γδ cells and the adhesion molecules CD44⁺, CD62L, LFA-1 and LPAM-1 indicated increase in CD4⁺ and B cells levels, with higher levels in blood than milk of group A, and significant expression of CD44⁺ in blood and milk and LPAM-1 in blood only. The CD8⁺ cells count in milk was higher than blood in the late stage. The peculiar feature of the early stage (group C) was the high level of γδ cells in the blood and milk, with tendency to express high level of CD62L. Compelling evidence could support the assumption that the dominant γδ cells at early stage of MAP infection could be of CD8CD2⁻WC+1⁺ phenotype. γδ cells appear as promising markers in defining early changes of MAP infection due to their important role in priming innate and cell mediated immunity. Possible utilization of these peculiar changes in the γδ cells level in the early diagnosis of MAP infection should be the subject of further research.     

Expansion of intracellular IFN-γ positive lymphocytes during Mycoplasma agalactiae infection in sheep

A method to assess the expansion of antigen-specific intracellular IFN-γ positive T cell subsets during the infection will be helpful for a better understanding of mycoplasmal infections physiopathology in the sheep. We analysed the percentage of antigen-specific lymphocytes positive for intracellular IFN-γ during the infection of sheep with Mycoplasma agalactiae by culturing peripheral blood mononuclear cells of infected or uninfected animals with irradiated M. agalactiae. The expansion of antigen-specific IFN-γ positive lymphocytes in infected sheep was initially sustained by CD4⁺ T cells at day 15 after infection, when antigen specific IgG start to be detectable, followed by CD8/IFN-γ double positive cells. γδ T-cells were not expanded at any time point analysed. IFNγ⁺ T cells disappear 60 days after infection, suggesting that antigen specific IFNγ⁺ T cells, mainly detected in the early phase of the disease, could be useful to understand the role of cell-mediated immunity during M. agalactiae infection.     

Potent immune responses induced by a Salmonella ghost delivery system that expresses the recombinant Stx2eB,FedF, and FedA proteins of the Escherichia coli-producing F18 and Shiga toxin in a murine model and evaluation of its protective effect as a porcine vaccine candidate

Background: In the pathogenicity of porcine edema disease (ED), which is caused by the Escherichia coli-producing F18 and Shiga toxin, F18⁺ fimbrial adhesins and Shiga toxin 2e (Stx2e) play pivotal roles in the colonization and enterotoxicity of this pathogen.

Objective: To develop a vaccine candidate against ED by combining three selected antigens of F18⁺ E. coli.

Methods: Genetically engineered Salmonella Typhimurium (ST) ghosts that express Stx2eB, FedF, and FedA were individually inserted in a ghost plasmid cassette, and the resultant plasmids were transformed into an attenuated ST (JOL912). The individual expression of Stx2eB, FedF, and FedA in JOL912 was validated by using an immunoblotting assay.

Results: Immunization of the ghosts in BALB/c mice led to a significant increase in antigen-specific secretory IgA and serum IgG. Significantly marked elevation of the CD3⁺CD4⁺ T cell subpopulation and lymphocyte proliferating activity in the primed splenocytes were also observed. Furthermore, mRNA of IL-4 and IFN-γ were highly upregulated in in vitro stimulated splenic T cells. Subsequently, the immunized mice showed significant protection efficacy against a lethal dose 50 of a virulent strain, resulting in approximately 85% and 92% survival rates in mice with a single- and double-dose immunization, respectively, compared to only 40% of the non-immunized controls.

Conclusion: A mixture of the ghosts expressing these three antigens is a potential vaccine candidate for protection against the porcine edema disease.     

Studies on the immune status of calves with chronic inflammation and thymus atrophy

The thymus is a primary lymphoid organ where the primary T cell repertoire is generated. Thymus atrophy is induced by various conditions, including infectious diseases, glucocorticoid treatment, and poor breeding management. Cattle with thymus atrophy tend to exhibit weak calf syndrome, a condition in which approximately half of neonates die shortly after birth. Calves with thymus atrophy that survive the first month typically contract chronic inflammatory diseases. In this study, we analyzed the populations of the peripheral blood mononuclear cells and thymocytes in calves with thymus atrophy. In addition, we evaluated polarization of master gene and cytokine mRNA expression in peripheral blood CD4⁺ cells in the calves. The population of CD4⁺CD8⁺ cells in thymus of the calves with thymus atrophy was lower than that of control calves. IL10 mRNA expression in peripheral blood CD4⁺ cells of calves with thymus atrophy was significantly lower than that of control calves. TBX21 mRNA expression in peripheral CD4⁺ cells of thymus atrophy calves was tended to be higher than that of the control group. In addition, FOXP3 mRNA expression in peripheral CD4⁺ cells of the thymus atrophy calves was tended to be lower than that of the control calves. Thymus atrophy calves exhibited chronic inflammatory disease leading, in severe situations, to conditions such as pneumonia with caseous necrosis. These severe inflammatory responses likely are due to decreases in IL10 mRNA expression, impairing control of macrophages, one of the main cell fractions of natural immunity.     

Effects of Waterborne Lead Exposure in Mozambique Tilapia: Oxidative Stress,Osmoregulatory Responses,and Tissue Accumulation

We studied the oxidative stress and osmoregulatory damage as well as the accumulation of lead in Mozambique Tilapia Oreochromis mossambicus exposed to different sublethal concentrations—low, medium, and high (0.5, 2.5, and 5.0 mg/L)—of waterborne lead for 14 d in a semistatic condition. The accumulated levels of Na⁺,K⁺-ATPase, glutathione (GSH), and thiobarbituric acid reactive substances (TBARS) were determined from samples of gill, liver, intestine, brain, kidney, and muscle tissues. At the end of the experiment, the GSH levels of most tissues were higher in the treated group than in the control group (especially in the liver and kidney) but lower in the intestine. The levels of TBARS in the gill and brain tissues of the fish exposed to high lead doses were significantly higher than those of fish in the control group. Na⁺,K⁺-ATPase activity seemed to be significantly inhibited in the gill, intestine, and brain tissues across all treatment groups. At the end of the study, the total amount of lead that had accumulated within the various tissues ranked as follows: intestines > kidney > brain > gill > liver > muscle. Our findings suggest that sublethal concentrations of lead can disrupt the health of Mozambique Tilapia and cause oxidative stress and osmoregulatory damage.

Received March 30, 2014; accepted November 28, 2014     

荷斯坦牛红细胞Na+ -K+ -ATP酶的研究进展

作者对荷斯坦牛红细胞Na⁺ -K⁺ -ATP酶的分子结构和基本特征、分布、作用机制、调控机制、生理功能、影响因素及其与热应激的关系进行了综述。     

Dynamics of CD3+ T-cell Distribution Throughout the Estrous Cycle and Gestation in the Bovine Endometrium

T cells are the dominant lymphocytes in the endometrium and are considered to play a crucial role in implantation and in the maintenance of gestation through cytokine production and immune regulation. The mechanisms underlying immunoregulation at the feto-maternal interface are still obscure for this complex system. Understanding the role of T cells is a key factor in understanding the endometrial immune system. In this study, the distribution of endometrial CD3⁺ T cells in bovines was examined by immunohistochemical analysis. The estrous cycle and gestation was divided into 4 stages, and the number of CD3⁺-positive T cells was counted in each stage. CD3⁺ cells were found in the endometrium in significant numbers throughout the estrous cycle and were mostly located in the subepithelial area. The number of CD3⁺ cells significantly increased in the early and mid-luteal phases but decreased after implantation with the progression of gestation. No T cells were found in the placentome or specifically in the tissues near the fetus, including the trophoblastic area. In addition, very few T cells were found in stromal regions close to the myometrium of the endometrium. These findings suggest that downregulation of bovine endometrial CD3⁺ T-cell functions is closely related to the successful maintenance of gestation in a spatiotemporal manner.     

Altered adenosine triphosphatase activities in pigs with naturally occurring hypertrophic cardiomyopathy

The purpose of this study was to determine whether myocardial adenosine triphosphatase (ATPase) activities were reduced in pigs with naturally occurring hypertrophic cardiomyopathy (HCM). The selection of hearts for the HCM and the normal control groups depended on histological examination. Specific ATPase activity and 5-nucleotidase activity were measured in left ventricular myocardium obtained from HCM (n=7) and normal control (n=7) animals. The histological features of HCM included marked disorientation of muscle cells, thickening of the intramural coronary arterial wall with a narrowed lumen, endocardial fibrosis and myocardial fibrosis. The HCM group showed significant increases in both heart weight (32%) and heart weight to body weight ratio (46%). The total ATPase activity in crude homogenates from the HCM group was significantly decreased by 16%. Azide-sensitive ATPase (mitochondrial ATPase) activity, ouabain-sensitive ATPase (Na⁺,K⁺-ATPase) activity, basal Mg²⁺-ATPase activity and Ca²⁺-ATPase activity were all significantly decreased by 18%, 30%, 20% and 50%, respectively. In contrast, no significant decrease was found in the mean values for 5-nucleotidase activity. These results suggest that myocardial ATPase activities are suppressed in pigs with naturally occurring HCMAbbreviations ATP adenosine triphosphate - gww grams wet weight - HCM hypertrophic cardiomyopathy - Pi inorganic phosphate     

Ouabain-sensitive respiration and protein synthesis in duodenal mucosa and liver in rats fed increasing levels of pea fibre or protein and housed in 18 or 28°C environments

Seventy‐two Wistar rats were used in two studies to investigate the effect of environmental temperature (18 or 28°C), and increasing levels of dietary fibre (low, 68 g/kg dry matter (DM); medium 110 g/kg DM; high, 157 g/kg DM) and protein (low, 91 g/kg DM; medium, 171 g/kg DM; high, 262 g/kg DM) on respiration attributable to Na⁺,K⁺‐ATPase activity and protein synthesis in duodenal mucosa and liver of rats. In vitro O₂ consumption in tissues was measured polarographically using a Clark‐style YSI biological O₂ monitor. Whole‐body O₂ consumption was measured with two open‐circuit respiration chambers. Whole‐body O₂ consumption was higher (p < 0.05) at 18°C than at 28°C. Rats fed the low protein diet had significantly higher (p < 0.05) whole‐body O₂ consumption than those fed the medium or high protein diet. Compared with 28°C, the environmental temperature of 18°C caused an increase (p < 0.05) in total O₂ consumption and O₂ consumption attributable to Na⁺,K⁺‐ATPase activity in duodenal mucosa. There was no effect (p > 0.05) of environmental temperature on total O₂ consumption, Na⁺,K⁺‐ATPase activity attributable to protein synthesis dependent on O₂ consumption in the liver. Total O₂ consumption and O₂ consumption attributable to Na⁺,K⁺‐ATPase activity increased (p < 0.05) in duodenal mucosa in rats fed the low level of dietary fibre compared with rats fed the medium level of dietary fibre. In vitro O₂ consumption determined in duodenal mucosa and in liver did not always correspond to whole‐body O₂ consumption. This may indicate that respiration in the duodenum and liver adapts differently and may not reflect changes in whole‐body respiration in response to dietary modification and changes in thermal environment.     

Optimized Transduction of Canine Paediatric CD34+ Cells Using an MSCV-based Bicistronic Vector

We have used a murine MSCV-based bicistronic retroviral vector, containing the common gamma chain (γc) and enhanced green fluorescent protein (EGFP) cDNAs, to optimize retroviral transduction of canine cells, including an adherent canine thymus fibroblast cell line, Cf2Th, as well as normal canine CD34⁺ bone marrow (BM) cells. Both canine cell types were shown to express Ram-1 (the amphotropic retroviral receptor) mRNA. Supernatants containing infectious viruses were produced using both stable (PA317) and transient (Phoenix cells) amphotropic virus producer cell lines. Centrifugation (spinfection) combined with the addition of polybrene produced the highest transduction efficiencies, infecting ∼75% of Cf2Th cells. An average of 11% of highly enriched canine CD34⁺ cells could be transduced in a protocol that utilized spinfection and plates coated with the fibronectin fragment CH-296 (Retronectin). Indirect assays showed the vector-encoded canine γc cDNA produced a γc protein that was expressed on the cell surface of transduced cells. This strategy may result in the transduction of sufficient numbers of CD34⁺ BM cells to make the treatment of canine X-linked severe combined immunodeficiency and other canine genetic diseases feasible. Suter, S.E., Gouthro, T.A., McSweeney, P.A., Nash, R.A., Haskins, M.E., Felsburg, P.J. and P.S. Henthorn, 2006. Optimized transduction of canine paediatric CD34⁺ cells using an MSCV-based bicistronic vector. Veterinary Research Communications, 30(8), 881–901     

Effects of dietary cation-anion balance on blood PH, acid base parameters, serum and urine mineral levels, and parathyroidhormone (PTH) in weanling horses.

This study demonstrated that the feeding of treatment diets with calculated dietary cation-anion balances (DCAB) of +370.43 (H) and -25.69 (L) did not have significant effects on blood pH, pCO₂, and HCO₃-. Serum Ca²⁺, P, Na⁺, and Cl^- as well as plasma PTH did not differ (P > .05) between the two treatment groups. Serum K⁺ was higher (P< .05) in horses fed diet H rather than diet L. The DCAB of the diet significantly affected urinary Ca²⁺, P, Na⁺, K⁺, and Cl^- excretion in the young growing horse. Urine Ca²⁺ and Cl- levels were higher (P < .01) in horses fed diet H versus diet L. Furthermore, levels of P, Na⁺, and K⁺ in the urine were higher (P < .01) in horses on diet H as opposed to diet L. Results of this study indicate that horses were able to maintain acid-base status regardless of diet. However, these data imply that growing horses consuming diets low in DCAB may be predisposed to abnormal bone mineralization due to the increase in calcium excretion which could lead to a weakening of the skeletal system.