Aromatic vitamin-dependent Salmonellae as vaccines in food animals: efficacy and persistence.
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Biomedical subjects
Publications and source records attributed to S C Smith.
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Mice rendered deficient in lymphotoxin (LT) by gene targeting in embryonic stem cells have no morphologically detectable lymph nodes or Peyer's patches, although development of the thymus appears normal. Within the white pulp of the spleen, there is failure of normal segregation of B and T cells. Spleen and peripheral blood contain CD4+CD8- and CD4-CD8+ T cells in a normal ratio, and both T cells subsets have an apparently normal lytic function. Lymphocytes positive for immunoglobulin M are present in increased numbers in both the spleen and peripheral blood. These data suggest an essential role for LT in the normal development of peripheral lymphoid organs.
The concept of the morphogenetic field has been used extensively in developmental biology. However, little is known about the mechanisms that partition these broad areas of tissue into the smaller areas which actually form the corresponding structures, and the remaining tissue. In the Mexican axolotl, the heart field forms as the anterior lateral plate mesoderm migrates over the underlying pharyngeal endoderm between stages 14 and 28. We have previously shown that both the mid-ventral and lateral walls of the pharyngeal cavity have considerable inductive capacity at stage 14. If this inductive capability, and the competence of the mesoderm to respond, is retained between stages 14 and 28, a much broader area of mesoderm would be induced than actually participates in heart development. In this paper, we use explant cultures to establish that pharyngeal endoderm retains its inductive activity, and that both pre-cardiac mesoderm and lateral plate mesoderm caudal to the pharyngeal cavity remain competent to respond to the induction throughout this period. We also map the specified region of the antero-lateral mesoderm between stages 14 and 28 by placing carefully measured areas of mesoderm in culture without inductive endoderm. We found that the region capable of initiating a spontaneous beat approximately doubles in size during this period. Since the specified region is larger than the actual heart primordium, some mechanism must exist to partition "induced" mesoderm into heart-forming and non-heart-forming areas.(ABSTRACT TRUNCATED AT 250 WORDS)
Chromosomal DNA restriction enzyme analysis and Southern blot hybridization were used to characterize Serpulina hyodysenteriae strains. When chromosomal DNAs from selected strains (reference serotypes) of S. hyodysenteriae were digested with the restriction endonuclease Sau3A and hybridized with a 1.1-kb S. hyodysenteriae-specific DNA probe, a common 3-kb band was always detected in S. hyodysenteriae strains but was absent from Serpulina innocens strains. When the chromosomal DNA was digested with the restriction endonuclease Asp 700 and hybridized with two S. hyodysenteriae-specific DNA probes (0.75 and 1.1 kb of DNA), distinct hybridization patterns for each S. hyodysenteriae reference strain and the Australian isolate S. hyodysenteriae 5380 were detected. Neither the 1.1-kb nor the 0.75-kb DNA probe hybridized with Asp 700- or Sau3A-digested S. innocens chromosomal DNA. The presence of the 3-kb Sau3A DNA fragment in S. hyodysenteriae reference strains from diverse geographical locations shows that this fragment is conserved among S. hyodysenteriae strains and can be used as a species-specific marker. Restriction endonuclease analysis and Southern blot hybridization with these well-defined DNA probes are reliable and accurate methods for species-specific and strain-specific identification of S. hyodysenteriae.
Sodium ion selective field effect transistors (ISFETs) were evaluated for their performance in measurement of sodium ions in whole blood for 'near patient' analysis in operating theatres and intensive care units. Performance was evaluated in comparison with a standard clinical laboratory sodium/potassium ion analyser (Radiometer KNA1) and with sodium and potassium assays using flame photometry on the plasma from each whole blood specimen. The imprecisions (coefficients of variation) of three ISFETs for sodium ion assay were 1.08, 1.56 and 1.10%, respectively. Robust bivariate linear regression (reweighted least squares preceded by least median of squares) of the ISFET versus KNA1 sodium ion activity yielded a regression coefficient of 1.08 and an intercept of -18.2 mM. The influence of potassium, protein and lipid on the measurement of sodium ions by both ISFETs and the KNA1 was assessed using robust multiple regression (also based on reweighted least squares preceded by least median of squares). In the regression versus flame photometry, protein was found to be more influential for the KNA1 (glass sodium ion selective electrode) than for the ISFET. Potassium had no influence on assays using the ISFET, but had a weak negative influence on assays using the KNA1. Two ISFETs lasted for more than 200 assays each demonstrating their robustness in the assay of whole blood.
Ophthalmologists employ electrophysiologic testing to diagnose and manage a variety of ocular conditions. The most common reason these tests are performed is to diagnose inherited vitreoretinal diseases. Patients of all ages may require electrophysiologic testing. The purpose of this article is to provide the ophthalmic registered nurse with a working knowledge of the three most commonly used electrophysiologic tests. We will provide background and practical information on how we perform electroretinography (ERG), electroculography (EOG), and dark adaptometry. Knowledge of the basics of this specialized testing will allow the ophthalmic nurse to provide more information for the patient. This knowledge will hopefully reduce patient anxiety and with better cooperation produce more accurate test results.
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OBJECTIVES: The goal of this study was to assess patients with end-stage heart disease after implantation of a left ventricular assist device at rest and during exercise compatible with activities of daily life. BACKGROUND: Mechanical circulatory assistance with a left ventricular assist device is an accepted therapy for bridging patients with end-stage heart disease to heart transplantation and has been proposed for long-term implantation. METHODS: Three patients (aged 37, 42 and 57 years) with end-stage heart failure required implantation of a pneumatically driven, asynchronous Thermedics left ventricular assist device while awaiting heart transplantation. All were assessed 1 month later during graded supine bicycle exercise (maximal work load 100 to 150 W). Detailed central hemodynamics, including continuous pulmonary artery oxygen saturation and oxygen consumption measurements, were obtained. Two of the patients also underwent upright treadmill exercise with oxygen consumption measurements. RESULTS: During supine bicycle exercise, the heart rate increased from 93 +/- 37 beats/min (95% confidence interval: mean +/- t0.025 x SE) at rest to 119 +/- 54 beats/min and left ventricular assist device rate increased from 82 +/- 47 to 109 +/- 55 beats/min. Oxygen consumption increased from 3.0 +/- 0.9 to 8.7 +/- 2.9 ml oxygen/min per kg body weight. Cardiac output increased from 6.0 +/- 4.4 to 9.6 +/- 7.1 liters/min, yielding an average exercise factor of 8.5 +/- 7.7 and an exercise index of 0.83 +/- 0.61. The patients assessed during treadmill exercise achieved a maximal oxygen consumption of 14.3 and 16.7 ml of oxygen/min per kg. No thromboembolic or other complications attributable to left ventricular assist device implantation occurred during the duration of support. All patients survived orthotopic heart transplantation and are doing well. CONCLUSIONS: Significant work loads compatible with activities of daily life and adequate exercise hemodynamics were demonstrated by these patients while awaiting heart transplantation. Definitive conclusions regarding the use of this device must be viewed as preliminary because only three patients were involved in this study and the failure rate may be as high as 71% (95% confidence interval of left ventricular assist device success as a bridge to transplantation 29.3% to 100%). Final conclusions regarding the safety and efficacy of the left ventricular assist device as a possible long-term circulatory support device must await results of larger multicenter trials in progress.
OBJECTIVES: Data from a national registry of 23 centers using cardiopulmonary support (CPS) were analyzed to compare the risks and benefits of prophylactic CPS versus standby CPS for patients undergoing high risk coronary angioplasty. BACKGROUND: Early data from the CPS registry documented a high angioplasty success rate as well as a high procedural morbidity rate. Because of this increased morbidity some high risk patients were placed on standby CPS instead of prophylactic CPS. METHODS: Patients in the prophylactic CPS group had 18F or 20F venous and arterial cannulas inserted and cardiopulmonary bypass initiated. Patients in the standby CPS group were prepared for institution of cardiopulmonary bypass, but bypass was not actually initiated unless the patient sustained irreversible hemodynamic compromise. RESULTS: There were 389 patients in the prophylactic CPS group and 180 in the standby CPS group. The groups were comparable with respect to most baseline characteristics, except that left ventricular ejection fraction was lower in the prophylactic CPS group. Thirteen of the 180 patients in the standby CPS group sustained irreversible hemodynamic compromise during the angioplasty procedure. Emergency institution of CPS was successfully initiated in 12 of these 13 patients in < 5 min. Procedural success was 88.7% for the prophylactic and 84.4% for the standby CPS group (p = NS). Major complications did not differ between groups. However, 42% of patients in the prophylactic CPS group sustained femoral access site complications or required blood transfusions, compared with only 11.7% of patients in the standby CPS group (p < 0.01). Among patients with an ejection fraction < or = 20%, procedural morbidity remained significantly higher in the prophylactic CPS group (41% vs. 9.4%, p < 0.01), but procedural mortality was higher in the standby group (4.8% vs. 18.8%, p < 0.05). CONCLUSIONS: Patients in the standby and prophylactic CPS groups had comparable success and major complication rates, but procedural morbidity was higher in the prophylactic group. When required, standby CPS established immediate hemodynamic support during most angioplasty complications. For most patients, standby CPS was preferable to prophylactic CPS during high risk coronary angioplasty. However, patients with extremely depressed left ventricular function (ejection fraction < 20%) may benefit from institution of prophylactic CPS.
The migration behavior of an homologous series of phenols in micellar electrokinetic chromatography is quantitatively presented. This model describes mobility in terms of fundamental physical and chemical constants of each solute (acid dissociation constant Ka, micelle binding constant Km), the pH of the buffer, and the micelle concentration ([M]) in the buffer. The model was used to predict the mobility of each solute over a two-dimensional pH/[M] space. Predicted and actual electropherograms show the usefulness of this technique.
Small peptides are often conjugated to large carrier proteins such as thyroglobulin to increase their immunogenicity. Antibodies are raised against peptide, thyroglobulin and contaminants, such as immunoglobulins or BSA in the thyroglobulin preparation, but anti-contaminant antibodies will not be revealed if immune serum is diluted in buffer containing immunoglobulin-contaminated BSA. This may lead to the development of an inappropriate hybridoma-screening assay which detects more anti-contaminant than anti-peptide antibodies if ELISA plates are coated with a capture antibody or blocked with BSA. Dilution of culture supernatants in buffer containing BSA and Tween 20 minimises the risk of false positive results and makes plate-blocking unnecessary. The high affinity peptide-specific antibodies in immune serum, which are more readily detected than lower affinity monoclonal antibodies, may result in an inadequately sensitive hybridoma-screening ELISA.
The migration behavior and optimum separation of a set of substituted phenols is presented. A mathematical model that describes mobility in CZE in terms of fundamental constants of each solute (acid dissociation constant Ka, mobility of the dissociated acid microA-) and the pH of the buffer was used to predict the mobility of each solute as a function of pH. The resolution between each peak pair was calculated, and a window diagram of minimum resolution as a function of pH was employed in determining the optimum pH. A comparison of the predicted and actual electropherograms shows the usefulness of this technique.
An immunohistochemical method using Pab1801, a monoclonal antibody specific to the human p53 protein, was applied to detect p53 expression in colorectal cancer and dysplasia complicating ulcerative colitis. Of 20 tissue samples with dysplasia, six showed positive immunoreactivity. Archival paraffin-embedded tissue blocks from 21 colitic cancers were analysed; 11 showed positive immunoreactivity, compared with ten of 21 samples from matched sporadic colorectal cancers (P not significant). Previous data suggesting that colorectal carcinoma complicating ulcerative colitis has a reduced frequency of c-Ki-ras mutation compared with sporadic cancer have led to the hypothesis that different genetic lesions underlie colitic and sporadic colorectal carcinoma. The present results suggest that this is not the case with regard to p53 gene alterations.
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In axolotl embryos homozygous for the cardiac-lethal (c) gene, the heart never begins to beat. Recently, we demonstrated that the c gene affects the heart mesoderm directly, making the latter incapable of responding to normal inductive stimuli. Based on these results, we proposed that a reaction-diffusion mechanism controls the later stages of vertebrate cardiogenesis. In the present study, we use a series of transplantations to examine the precardiac mesoderm of wild-type and cardiac-lethal mutant axolotl embryos for the activation and inhibition predicted by our model. When one or both heart primordia in c/c embryos were replaced with +/+ tissue at early stages, functional hearts developed, indicating that an activator-like activity is present in wild-type heart mesoderm. At later stages, replacement of the entire heart-field mesoderm also rescued mutant embryos, but replacement of only the heart-forming mesoderm did not, indicating that the mutant heart field has inhibitory properties. These results demonstrate that activator- and inhibitor-like factors are present in pre-cardiac mesoderm and thus provide evidence for reaction-diffusion control of postinductive heart development.
A monoclonal antibody to Serpulina hyodysenteriae 8930 was produced and was used to probe pronase-treated cell lysates of S. hyodysenteriae isolates in immunblots. The results showed that the monoclonal antibody was specific for only five closely related S. hyodysenteriae isolates: 8930, 5380, 70A, RMIT 88, and RMIT 97.
Two DNA probes, one 1.1- and one 0.75-kb probe, specific for Serpulina hyodysenteriae were isolated from a genomic library generated from virulent S. hyodysenteriae 5380. These probes are highly specific and react with all S. hyodysenteriae strains tested. Under stringent conditions, the DNA probes did not react with the nonpathogenic species Serpulina innocens or with other species of enteric bacteria, including Escherichia coli. Both probes are able to detect S. hyodysenteriae in colony blot hybridizations, and when applied to fecal specimens, they can detect 10(4) S. hyodysenteriae cells in 0.1 g of seeded fecal matter. Both probes can detect S. hyodysenteriae in fecal specimens from swine with clinical signs of swine dysentery after experimental challenge and from swine from a herd with an acute outbreak of swine dysentery. These probes have application as a diagnostic tool in veterinary microbiology.
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