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Biomedical subjects

W T Dixon

Publications and source records attributed to W T Dixon.

At least 19 recordsLinked to original sources
PubMed2001

Feed restriction and insulin treatment affect subsequent luteal function in the immediate postovulatory period in pigs: progesterone production in vitro and messenger ribonucleic acid expression for key steroidogenic enzymes.

Progesterone production and release in vitro, and mRNA expression for key steroidogenic enzymes, were studied in luteal tissue recovered in the immediate postovulatory period from cyclic gilts allocated to one of three treatments: moderate feed restriction during the first (RH) or second week of the estrous cycle, with (HR+I) or without (HR) concomitant injections of long-acting insulin. Time of feed restriction affected neither progesterone production or release, nor mRNA expression for several key steroidogenic enzymes. However, luteal tissue from RH but not from HR gilts responded to LH stimulation by increasing progesterone production and release (P: < 0.05). Insulin treatment increased progesterone production and release, restored luteal tissue responsiveness to LH, up-regulated steroidogenic enzyme mRNA expression, and down-regulated the tissue inhibitor of metalloproteinase-I mRNA expression in HR+I compared with HR gilts (P: < 0.05). In vitro progesterone production and gene expression were affected by time of tissue collection after ovulation in RH and HR gilts but not in HR+I gilts, and were correlated with temporal changes in oviductal and peripheral plasma progesterone concentrations. Inherent differences in luteal function therefore appear to mediate latent effects of nutrition and insulin treatment on circulating progesterone concentrations in the critical postovulatory period in gilts.

3-Hydroxysteroid Dehydrogenases↗
PubMed2001

Coordinate expression of matrix-degrading proteinases and their activators and inhibitors in bovine skeletal muscle.

Matrix metalloproteinases (MMP) responsible for degradation of connective tissue are found in most tissues. The MMP are regulated at the levels of transcription, zymogen activation by plasmin or membrane-type- (MT) MMP, and control of enzyme activity by tissue inhibitors of metalloproteinases (TIMP). Whole bovine skeletal muscle showed multiple MMP activities on gelatin zymography and also expressed mRNA encoding MMP-1, -2, -9, -14, and -16, tissue inhibitors of metalloproteinase (TIMP)-1, -2, and -3 and plasminogen activator and its receptor. Purified intramuscular fibroblasts and myogenic cell culture derived from satellite cells expressed most or all of these elements. Statistical analysis (n = 35) revealed a strong positive correlation among the mRNA levels of several elements of the MMP system, including MMP-2, MMP-14, TIMP-1, -2, and -3 (r = 0.614 to 0.930, P < 0.0001). Our results provide an extensive profile of an extracellular proteolytic cascade involving MMP in skeletal muscle and suggest that 1) the activation cascades of muscle MMP may be initiated by both plasmin and membrane-type MMP; 2) a group of genes involved in the same "arm" of zymogen activation are coexpressed in this tissue; and 3) skeletal muscle cells, in addition to the intramuscular fibroblasts, express an extensive complement of MMP and related proteins.

Animals↗
PubMed2000

Western blotting for detection of glucocorticoid receptors in the brain and pituitary gland from adrenal intact pigs.

The objectives of the experiments presented here were to develop a method for measuring total glucocorticoid receptors (GRs), in adrenal intact pigs, and to describe the distribution of this receptor in the pituitary gland and selected brain regions. Six male pigs, 14 days of age, were euthanized and the pituitary gland, hypothalamus, prefrontal cortex, and hippocampus were prepared for western blotting using a traditional soluble cytosolic preparation or a whole cell extract preparation. Samples were electrophoresed and immunoblotted using the anti-human GR antibodies #57 and #51. The #57 antibody labeled a doublet at 95 and 87 kDa whereas the #51 GR antibody labeled a single band at 87 kDa. Affinity labeling of GR with [(3)H] dexamethasone mesylate revealed a single band at 87 kDa. Using the soluble cytosolic preparation GR concentrations were highest in the pituitary gland followed by the hypothalamus (P<0. 0001). The amount of GR in the hippocampus and prefrontal cortex was equivalent but lesser (P<0.0001) than that found in the pituitary gland or hypothalamus. With the whole cell extract tissue preparation, GR concentrations were highest (P<0.05) in the prefrontal cortex and hippocampus with lower (P<0.05) levels detected in the hypothalamus. Taken together, these results indicated that the 87-kDa band was the functional GR and that regional distribution of GRs is specific to the type of tissue preparation used. In addition, we have described a procedure (from whole cell extracts) for measuring total GRs in neural and nonneural tissues from adrenal intact pigs.

Adrenal Glands↗
PubMed2000

The effects of mutated skeletal ryanodine receptors on calreticulin and calsequestrin expression in the brain and pituitary gland of boars.

Mutations in skeletal ryanodine receptors (sRyR) result in malignant hyperthermia in humans and Porcine Stress Syndrome (PSS) in pigs. Whether the sRyR is expressed in neuronal tissue and what impact it has on neuronal function is relatively unexplored. We have hypothesized that the presence of mutated sRyR may be accompanied by compensatory changes in Ca(2+)-binding protein expression. We were interested in whether pigs heterozygous for mutated sRyR would show changes in the expression of Ca(2+)-binding proteins, in specific regions of the brain, and whether changes in this expression would be accompanied by the presence of sRyR within that region. The objectives of the current experiments were to determine (1) whether calreticulin (CR) and calsequestrin (CS) are expressed in the pituitary gland and brain of the pig, (2) if boars heterozygous for mutated sRyR differed from wild-type boars in the expression of CR or CS, and (3) if altered Ca(2+)-binding protein expression would be accompanied by the presence of sRyR mRNA. Boars either heterozygous or wild-type (n=6) for the mutation in sRyR known to cause PSS, were euthanized and the pituitary gland and brains were collected for western blotting for CR and CS. An additional four wild-type boars were sacrificed and brains were collected for in situ hybridization for sRyR mRNA. Immunoreactive CR was expressed in porcine tissues with highest (p<0.0001) expression in the pituitary gland and lower but equivalent expression in the hypothalamus, frontal cortex, and hippocampus. Immunoreactive CS was not detectable in the pituitary gland while low levels were observed in the hypothalamus and frontal cortex. Dramatically higher (p<0.0001) levels of CS were found in the hippocampus. Genotype did not affect CR expression in the pituitary gland or any brain region examined. Immunoreactive CS levels were lower (p<0.002) in the hippocampus of heterozygous compared to wild-type boars. In situ hybridization experiments revealed the presence of sRyR mRNA in the hippocampus equally distributed across all cell subfields. In conclusion, both CR and CS were expressed in the porcine brain with specific patterns of expression across the brain regions examined. Boars heterozygous for mutated sRyR had lower CS in the hippocampus, which was accompanied by the expression of mRNA for sRyR.

Animals↗
PubMed2000

The metabolic availability of vitamin A is decreased at the onset of diabetes in BB rats.

Streptozotocin (STZ)-induced diabetic rats have been associated with an impaired metabolic availability of vitamin A (retinol). This study was undertaken to investigate whether Biobreeding (BB) rats, in which diabetes mellitus resembling human type I diabetes develops spontaneously, respond the same way at the onset of diabetes. Weaning diabetes-prone (BBdp) and normal (BBn) BB rats consumed NIH-07 nonpurified diet ad libitum until 120 d of age. Plasma and hepatic concentrations of retinol and its carriers, retinol-binding protein (RBP) and transthyretin (TTR) were lower in diabetic BB (BBd) rats than in BBn rats. In parallel with RBP, the abundance of mRNA was lower in the liver of BBd rats. Furthermore, the status of zinc, an important factor for the synthesis of RBP, was also disturbed in BBd rats, as indicated by lower circulatory levels and greater urinary excretion. To determine whether the biochemical evidence of vitamin A deficiency in BBd rats could be reversed, BBdp rats were fed a diet supplemented with vitamin A either alone or in combination with zinc. None of these treatments increased plasma vitamin A concentration. The hepatic abundance of RBP mRNA was significantly greater, whereas circulatory RBP concentrations were unaffected by vitamin A plus zinc supplementation. Overall, these results suggest that impaired metabolic availability of vitamin A, possibly caused by its decreased transport from hepatic stores, is another metabolic derangement associated with type I diabetes.

Animals↗
PubMed2000

Neonatal handling permanently alters hypothalamic-pituitary- adrenal axis function, behaviour, and body weight in boars.

Neonatal handling permanently alters hypothalamic- pituitary-adrenal axis (HPA) function in rats. In the rat, this treatment increases hippocampal glucocorticoid receptors (GR) and dampens plasma ACTH and corticosterone responses to stressors. The objectives of this study were to determine whether neonatal handling of pigs would effect permanent changes in plasma corticosteroid binding capacity (CBG), basal or stressor-induced plasma cortisol and ACTH concentrations, brain or pituitary GR levels, dexamethasone suppression of plasma cortisol and ACTH concentrations, behaviour in an open field-test pen, and body weights. Twelve litters of pigs were randomly assigned to either neonatal handling or no disturbance. Handled litters were removed from the farrowing crate for 10 min per day for the first 14 days of life. Male pigs were kept for the study and the boars were weighed monthly. At 7 months of age, boars were tested for locomotory behaviour in an open field-test pen. The boars were implanted with indwelling ear-vein catheters and blood samples were obtained basally, during and after application of a nose snare, and after 0.04 mg/kg dexamethasone. Boars were killed and blood samples were obtained and the brain and pituitary glands collected. Handled boars had greater (P<0.05) plasma CBG binding and lower basal total (P<0.05) and calculated free (P<0.03) plasma cortisol concentrations. No significant differences between treatments were found in plasma ACTH or cortisol responses to a nose-snare stressor; however, when killed, handled boars had greater (P<0.02) plasma ACTH concentrations. Handled and non-handled boars did not differ in plasma ACTH or cortisol responses to dexamethasone. There was no treatment effect on GR expression in the pituitary gland, frontal cortex, hippocampus, or hypothalamus. Behaviourally, the handled boars had higher (P<0.03) locomotor scores over inner squares and a lower (P<0.05) ratio of outer:inner squares entered in open field-tests. During the first 7 months of life, body weights were lower (P<0.004) for handled boars. In conclusion, neonatal handling permanently altered HPA function in pigs, but in a manner dissimilar to that found in the rat. These changes induced in the pig were not beneficial for commercial production with respect to body weight.

Adrenocorticotropic Hormone↗
PubMed2000

The effects of mutated skeletal ryanodine receptors on hypothalamic-pituitary-adrenal axis function in boars.

The objectives of the current experiment were to determine whether boars heterozygous for the mutation in skeletal ryanodine receptors (sRyR), known to cause porcine stress syndrome, differed from wild-type boars in hypothalamic-pituitary-adrenal axis (HPA) function. We have examined basal plasma ACTH, cortisol, and corticosteroid-binding globulin (CBG) concentrations; plasma ACTH and cortisol responses to a nose-snare stressor and at slaughter; dexamethasone suppression of plasma ACTH and cortisol concentrations; and glucocorticoid receptor (GR) density in the pituitary gland, hippocampus, hypothalamus, and frontal cortex. We have also examined carcass yields, composition, and meat quality to determine whether differences in HPA activity were accompanied by an increased incidence of meat quality characteristics associated with pale, soft, exudative (PSE) meat. Thirty boars either heterozygous or wild-type (n = 15 per genotype) for mutated sRyR were tested for HPA function at 7 mo of age. Heterozygous boars had lower basal plasma ACTH (P < .05) and cortisol (P < .04) concentrations. Integrated basal plasma ACTH and cortisol levels were also lower (P < .05 and P < .005, respectively). Genotype had no significant effect on basal CBG, stressor-induced (nose snare or slaughter) or dexamethasone suppression of plasma ACTH or cortisol concentrations. No differences in immunoreactive GR levels were found in the pituitary gland or any brain region examined. We did find a significant, negative correlation (r = -.62, P < .02) between peak (0800) basal plasma ACTH concentrations and hippocampal GR levels. The alterations in basal HPA function in heterozygous boars were accompanied by lighter body weights (P < .03), decreased carcass fat depth (P < .04), and increased carcass lean yields (P < .02). There was a higher incidence of meat quality characteristics associated with PSE meat in heterozygous boars indicated by higher carcass temperatures (P < .04) and meat brightness (P < .0001) with lower carcass pH at slaughter (P < .03) and after chilling (P < .003). In conclusion, we have found differences in basal and not stressor-induced HPA function between boars heterozygous and wild-type for mutated sRyR. This altered basal HPA activity was accompanied by an increased incidence of meat quality aspects associated with PSE meat in heterozygous boars.

Adrenocorticotropic Hormone↗
PubMed1999

MRI techniques for cardiovascular imaging.

Over the last several years, cardiovascular MRI has benefited from a number of technical advances which have improved routine clinical imaging techniques. As a result, MRI is now well positioned to realize its longstanding promise of becoming the comprehensive cardiac imaging test of choice in many clinical settings. This may be achieved using a combination of basic advanced techniques. In this overview, the basic cardiac MRI techniques which are clinically useful are reviewed, and the recent technical advances which are clinically promising are described. These advances include routine black blood and cine bright blood techniques that are high speed (<10s per black blood image or cine slice), multislice whole heart perfusion imaging methods, and recently emerging real-time imaging methodologies. J Magn. Reson. Imaging 1999;10:590-601.

Cardiovascular System↗
PubMed1999

Segmented spin-echo pulses to increase fMRI signal: repeated intrinsic diffusional enhancement.

Since its inception, functional magnetic resonance imaging (fMRI) has seen rapid progress in the application to neuroscience. Common gradient-recalled acquisition methods are susceptible to static field inhomogeneities, resulting in signal loss at the medial temporal area important for memory function or at the basal ganglia area for motor control. In addition, they are susceptible to the contaminating signals of large vein origin, such as the signals from its surrounding cerebrospinal fluid (CSF) leading to false-positive activation. Spin echoes overcome these drawbacks. However, they are less sensitive to blood oxygenation level dependent (BOLD) susceptibility changes because of their refocusing mechanism. A method is presented here to enhance the spin-echo fMRI signal by recruiting more spins to participate in the dynamic BOLD process. This method divided a conventional T(2) weighting period into several segments separated by blocks of extra free diffusion time. Before the extra diffusion time spins are restored to the longitudinal axis preventing rapid transverse relaxation. This process allows more spin access to the regions that experience the BOLD field gradient. Because of the increased spin population that is modulated by the capillary BOLD field gradient, the functional signal is increased. Spin-echo echo-planar imaging (EPI) with this enhancement may be a useful technique for fMRI studies at inhomogeneous areas such as the air/tissue interface. Magn Reson Med 42:631-635, 1999.

Brain↗
PubMed1999

Lack of class I major histocompatibility antigens on trophoblast of periimplantation blastocysts and term placenta in the pig.

In this study, the pattern of expression of class I major histocompatibility (MHC) antigens and mRNA on periimplantation blastocysts and term placental tissue was determined for the pig. Class I MHC antigens could not be detected immunohistochemically either on extra-embryonic membranes or on the embryonic portion of Day 14, 16, 22, and 25 blastocysts. Nor could class I MHC antigens be detected on the outer trophoblast epithelium and inner endodermal surface of the chorioallantoic membrane or on the outer and inner surfaces of the amnion at term. However, MHC class I antigens were detected on the vascular mesoderm found in both the chorion and amnion at term, and in Day 25 extra-embryonic membranes. Uterine endometrial cells and tissues and maternal peripheral blood leukocytes stained strongly for class I MHC antigens. There was a large difference in the intensity of class I MHC mRNA signal, detected by Northern blot analysis, in embryo/fetus-derived tissues compared to that in maternal tissues. The embryos appeared to express even less class I MHC mRNA than did the extra-embryonic membranes. In addition, in situ hybridization of Day 16 blastocysts indicated class I MHC mRNA to be ubiquitously expressed at low levels in embryos and extra-embryonic tissues compared to uterine endometrial tissue controls. Taken together, these results indicate that class I MHC antigens are either not expressed on the surface of the extra-embryonic/fetal membranes during gestation in the pig or are expressed at very low levels, and that specific mRNA is expressed at correspondingly low levels.

Allantois↗
PubMed1998

Magnetic resonance coronary angiography using navigator echo gated real-time slice following.

Navigator echo gating allows for the elimination of breath-holding in MR imaging by providing a real-time monitor of respiratory position to gate image acquisition. In this study we examined the advantages and utility of real-time, navigator echo gated slice following technique in 2D magnetic resonance coronary angiography of patients with coronary artery disease. Thirteen patients with coronary artery disease were examined. MR images of the right coronary artery (RCA) were obtained parallel to the atrioventricular groove to image long sections of the RCA in a small number of slices. In-plane resolution was 0.7 x 0.9 mm and 2-6 signals were averaged to support this high spatial resolution. Targeted maximum intensity projection (MIP) images were generated from the slices to present the RCA in a single image. All patients had x-ray angiograms available for comparison with the MR images. Using the navigator echo gated real-time slice following technique, MRI successfully obtained images in 11 of 13 cases. The technique failed in two patients with irregular breathing patterns. The average length of the RCA seen in the 11 successful MR exams was 61 mm and the average length seen in the x-ray angiograms was 80 mm. Eight patients were determined to be without disease in the RCA by x-ray angiography, and all eight were correctly identified as normal on the MRI exam. In the three patients who had a successful MRI exam and were determined to have disease in the RCA by x-ray angiography, MRI identified the lesion in two cases. In the third case MRI indicated a discrete lesion and x-ray angiography indicated diffuse disease without a focal lesion. Navigator echo gating improves patient tolerance, provides aligned sections of coronaries over multiple slices, and allows for improved resolution through signal averaging. This preliminary patient study suggests that navigator echo gated magnetic resonance coronary angiography may play a role in evaluating coronary artery disease.

Coronary Angiography↗
PubMed1996

Myocardial suppression in vivo by spin locking with composite pulses.

Improved myocardium-blood contrast by myocardial suppression resulting from T1 rho-weighting in contrast-enhanced, gradient-echo, bright-blood cine images, acquired at 1.5T, is shown. In the standard images, blood has twice the intensity of muscle. In similar T1 rho-weighted images, it has 3-4 times the intensity of muscle. A composite spin-lock pulse before each observation pulse provides T1 rho-weighting. A typical pulse was: 90y-135x-360x-135x-90(-y) with element durations: 0.84, 1.26, 8.12, 1.26, and 0.84 ms. The tolerance of this composite pulse to shimmering and frequency errors allows spin locking with comparatively weak RF and therefore low specific absorption rate (SAR). Initial clinical evaluation on patients with poor ventricular function demonstrates both a qualitative and quantitative improvement in delineation of myocardial borders.

Absorption↗
PubMed1996

Localization and gene expression of glucose transporters in bovine mammary gland.

Glucose uptake in the mammary gland is a rate-limiting step in milk synthesis. To study glucose transporters in the bovine mammary gland, the erythrocyte-type glucose transporter (GLUT1) and the insulin-responsive glucose transporter (GLUT4) proteins were assessed by Western blotting and immunohistochemical staining, using polyclonal antibodies against the C-terminal peptide of GLUT1 and GLUT4. Our results demonstrated that the bovine mammary gland expressed a relatively high level of GLUT1 protein, whereas GLUT4 protein was not detected in the mammary gland of either lactating or dry cows. The absence of GLUT4 may indicate that glucose transport is not regulated by insulin in the lactating and dry bovine mammary gland. The anti-GLUT1 antibody strongly stained the single layer of epithelial cells of mammary alveoli. The expression of GLUT1 mRNA was similar in the mammary gland of late lactation and non-lactating cows. However, a smaller molecular weight species (38 kDa) of GLUT1 protein was detected in the mammary gland of non-lactating cows where its abundance in crude membrane preparation was 80% higher than in lactating animals. There were no significant differences in GLUT1 mRNA in bovine mammary gland at 118 d and 181 d postpartum, however, GLUT1 protein expression tended to be greater at 118 d postpartum.

Animals↗
PubMed1996

Two-dimensional coronary MR angiography without breath holding.

PURPOSE: To determine whether breath holding can be eliminated in two-dimensional magnetic resonance (MR) imaging of the coronary arteries by using real-time respiratory gating. MATERIALS AND METHODS: Thirty-one subjects (20 healthy volunteers, 11 patients) underwent MR imaging. In 13 subjects, a respiratory monitoring belt was used, and in 18 subjects, a navigator echo was used. MR imaging was performed with breath holding, respiratory gating, and respiratory gating with two signals acquired. Three reviewers conducted a blinded review of the images, and overall image quality was rated on a scale from 1 (poor) to 5 (excellent). RESULTS: Respiratory gating with two signals acquired provided image quality superior to that with breath-hold imaging (3.7 vs 3.0, respectively; P < .05). Measurements of signal-to-noise ratio (14.5 for respiratory gating with two signals acquired and 11.9 for breath holding) supported the results of the image review. Navigator-echo gating provided better image quality than the monitoring belt (3.7 vs 3.1, respectively; P < .05). CONCLUSION: Breath holding may be eliminated by gating image acquisition to a real-time monitor of respiratory position. Respiratory gating enables improved resolution by means of acquisition of multiple signals, provides aligned sections of coronary arteries, and improves patient tolerance.

Adult↗
PubMed1996

Immune responses in chickens against lipopolysaccharide of Escherichia coli and Salmonella typhimurium.

Immunization of chickens with whole bacteria results in the production of antibodies specific to lipopolysaccharide (LPS), a major constituent of the outer membrane of Gram-negative bacteria. However, there is relatively limited information available concerning immune response of purified LPS in this species. In the present study, immune responses were examined in serum and egg yolk from two groups of chickens injected with entire LPS from Escherichia coli and lipid A free LPS from Salmonella typhimurium. The results demonstrated that the increase of antibody activity occurs first in serum, and then in egg yolk with a lag in time of 1 to 3 wk in both groups of chickens. However, the time of elevated levels of antibodies activity was much shorter in chickens immunized with S. typhimurium LPS (< 1 wk) than in those immunized with E. coli LPS (4 wk). A lack of lipid A is the S. typhimurium antigen may be a factor related to this difference.

Animals↗
PubMed1995

The effect of inhomogeneous sample susceptibility on measured diffusion anisotropy using NMR imaging.

Water diffusion measurements in white matter of freshly excised pig spinal cord and in parenchyma of fresh celery (excluding the fibers along the edge of the stalk) were performed using NMR at 200 MHz. In white matter of pig spinal cord, the measured diffusion coefficient is anisotropic and independent of sample orientation with respect to the magnetic field. In celery parenchyma, diffusion is isotropic and independent of orientation in the magnetic field when using a diffusion sequence that gives results independent of self-induced magnetic-field gradients. However, when the standard diffusion pulse sequence that gives results dependent upon self-induced magnetic-field gradients is used, diffusion in celery appears isotropic when the stalk is oriented parallel to the magnetic field but anisotropic when oriented perpendicular. Susceptibility variations leading to anisotropic self-induced magnetic-field gradients approximately 3 kHz/cm in magnitude when the celery is oriented perpendicular to the magnetic field can explain this apparent anisotropic diffusion. A study of the apparent diffusion coefficient (ADC) in celery as a function of diffusion times ranging from 8 to 22 ms indicates that the motion is at most only slightly restricted. Therefore, although the effect is not seen in all types of samples, one must be aware that self-induced gradients may affect the ADC and may cause isotropic diffusion to appear anisotropic. In addition, NMR experiments that change diffusion-sensitizing gradient timings to study restricted diffusion change the effects of the self-induced gradients as well as the effect of barriers on the ADC, complicating interpretation.

Algorithms↗
PubMed1995

Divalent cations control cell-substrate adhesion and laminin expression in normal and malignant human melanocytes in early and late stages of cellular differentiation.

Integrins are a class of adhesion molecules that depends on divalent cations for proper function. This study examined whether human normal melanocytes and malignant (metastatic) melanocytes with early and late stages of cellular differentiation (G361 and SK-MEL-23, respectively) would differ in integrin-mediated adhesion to fibronectin, laminin, as well as collagens type I and type IV, and whether divalent cations could influence the strength of adhesion ability. Integrin subunit expression was determined by flow cytometry using integrin subunit-specific antibodies as probes. Integrin-specific adhesion was determined using soluble glycine-arginine-glycine-asparagine-serine peptide and integrin subunit-specific antibodies as functional blocking agents. This study shows that both normal and malignant melanocytes adhere to extracellular matrices in a divalent cation-dependent manner, and adhesion strength varies with the cation species. Integrins can be rapidly activated by small alterations in cation concentration, manganese being the most potent. There were marked differences in substrate adhesion between normal melanocytes and metastatic malignant melanoma cells, but these differences were not related to the stage of cellular differentiation. All the three cell types, however, expressed the same integrin subunits at approximately the same levels. This suggests that substrate adhesion of melanocytes and melanoma cells might involve some integrin-independent mechanisms as well. Manganese, in particular, appears to cause adhesion by activating both integrin-dependent and -independent mechanisms.

Amino Acid Sequence↗