Development of a Gene Expression Assay for the Diagnosis of Mycobacterium bovis Infection in African Lions (Panthera leo).

Mycobacterium bovis infection, the cause of bovine tuberculosis (BTB), is endemic in wildlife in the Kruger National Park (KNP), South Africa. In lions, a high infection prevalence and BTB mortalities have been documented in the KNP; however, the ecological consequences of this disease are currently unknown. Sensitive assays for the detection of this infection in this species are therefore required. Blood from M. bovis-exposed, M. bovis-unexposed, M. tuberculosis-exposed and M. bovis-infected lions was incubated in QuantiFERON® -TB Gold (QFT) tubes containing either saline or ESAT-6/CFP-10 peptides. Using qPCR, selected reference genes were evaluated for expression stability in these samples and selected target genes were evaluated as markers of antigen-dependent immune activation. The abundance of monokine induced by gamma interferon (MIG/CXCL9) mRNA, measured in relation to that of YWHAZ, was used as a marker of ESAT-6/CFP-10 sensitization. The gene expression assay results were compared between lion groups, and lenient and stringent diagnostic cut-off values were calculated. This CXCL9 gene expression assay combines a highly specific stimulation platform with a sensitive diagnostic marker that allows for discrimination between M. bovis-infected and M. bovis-uninfected lions.

Management of spontaneous enophthalmos due to silent sinus syndrome: a case report.

Silent sinus syndrome (SSS) is a rare disease exhibiting unilateral enophthalmos and hypoglobus. A 26-year-old white female presented with right side enophthalmos and hypoglobus. There was no history of previous trauma or maxillary sinus diseases. A CT scan showed an opacified right maxillary antrum with decreased volume and downward bowing of the right orbital floor. From clinical and radiological findings the diagnosis SSS was made. Biopsies were collected from the maxillary sinus for the exclusion of malignancy. Two months later orbital floor reconstruction was carried out. Before antrostomy of the affected maxillary sinus, a relative enophthalmos of 4mm was determined. Five days after antrostomy the value reduced to 2.3mm. During the following 2 months the enophthalmos remained constant. At the end of the operation for orbital floor reconstruction it was 0.1mm. Five days after surgery the relative enophthalmos increased to 0.8mm. The value remained constant during the following 3 months. Initial antrostomy of the affected maxillary sinus may lead to a relevant, spontaneous reduction of enophthalmos. After a minimum period of 2 months a re-evaluation should be made, if a reconstruction of the orbital floor is still necessary for the correction of the globe position.

Effect of bimakalim (EMD 52692), an opener of ATP sensitive potassium channels, on infarct size, coronary blood flow, regional wall function, and oxygen consumption in swine.

OBJECTIVE: The aim was to assess whether bimakalim, an opener of ATP sensitive potassium channels, can reduce infarct size in swine myocardium. METHODS: Experiments were performed in open chest pigs subjected to a 60 min occlusion of a branch of the left anterior descending coronary artery and to 2 h reperfusion. Five groups of animals were studied. In seven animals bimakalim infusion (3 micrograms.kg-1 bolus over 5 min followed by 0.1 microgram.kg-1.min-1) was started at 45 min of coronary occlusion and continued until 60 min of reperfusion (group A), while in seven other animals the bimakalim infusion was started 15 min before occlusion and also ended at 60 min of reperfusion (group B). In a further seven animals bimakalim infusion was started 15 min before coronary occlusion, but was stopped at the onset of ischaemia (group C). In the fourth group of animals (n = 7), a hydralazine infusion (0.2 mg.kg-1 over 15 min) was started 15 min before the occlusion and also terminated at the start of occlusion. The dose of hydralazine was chosen such that it lowered arterial pressure to the same extent as bimakalim. A fifth group of animals (n = 7) received the vehicle and served as controls. At the end of the protocol, infarct size (as percent of risk region) was determined by incubating myocardium with p-nitrobluetetrazolium. Regional myocardial oxygen consumption (MVO2) was calculated as the product of coronary blood flow (electromagnetic flowmeter) and the difference in the oxygen contents of the aorta and the interventricular vein accompanying the left anterior descending coronary artery. Regional wall function was quantified with ultrasonic crystals aligned to measure wall thickening (% delta WT). RESULTS: In all pigs in which bimakalim treatment was started prior to the 60 min coronary occlusion, infarct size was significantly reduced [B: 22.4(SEM 4.5)%; C: 35.3(6.6)%] compared with 60.4(5.2)% in pigs subjected to 60 min of ischaemia only (p < 0.05); drug-induced potassium channel opening during reperfusion had no effect [A: 56.6(4.1)%]. Treatment with hydralazine did not reduce infarct size [59.4(4.3)%]. Neither drug altered % delta WT; however, they reduced MVO2 by 36.5% in B, by 27.1% in C, and by 14.6% in the hydralazine group. CONCLUSIONS: Bimakalim treatment prior to the onset of a 60 min coronary occlusion increases the tolerance of pig myocardium to ischaemia. The data are consistent with the hypothesis that bimakalim reduces infarct size by activation of cardiac ATP sensitive potassium channels and not through unloading of the heart because of its vasodilator effects.

Prognostic value of immunohistochemically detected CD44 isoforms CD44v5, CD44v6 and CD44v7-8 in human breast cancer.

We investigated the expression of CD44 isoforms containing variant exons v5, v6 and v7-8 in 115 human breast cancer specimens by means of immunohistochemistry. CD44 isoforms CD44v5, CD44v6 and CD44v7-8 were detected in 56% (n = 64), 24% (n = 28) and 15% (n = 17), respectively. In 36 specimens of axillary lymph node metastasis, expression of CD44v5, CD44v6 and CD44v7-8 was found in 94% (n = 34), 92% (n = 33) and 89% (n = 32), respectively. Five year survival rates with or without CD44v5 and CD44v6 expression were 71% versus 86% (log-rank test, P = 0.02) and 62% versus 81% (log-rank test, P = 0.001), respectively. For disease-free survival, expression of CD44v5, CD44v6 and CD44v7-8 showed a prognostic impact (log-rank test, P = 0.004, P = 0.0001 and P = 0.0001, respectively). However, multivariate analysis revealed that all investigated CD44 isoforms failed to be independent predictors of the patient's outcome.

Posterior pituitary ectopy in children with idiopathic growth hormone deficiency.

AIMS: To evaluate the underlying pathogenesis in children with pituitary hormone deficiency by means of high resolution MRI of the brain. PATIENTS/METHODS: Thirty-seven children with short stature and isolated GH deficiency (IGHD, n = 17) or multiple pituitary hormone deficiency (MPHD, n = 20) were subjected to an MRI of the brain at the age of 1.0-17.3 years. The anatomic condition of the hypothalamo-pituitary area was analyzed and the height of the pituitary gland was measured and compared to the data of age-matched healthy subjects. RESULTS: Seventy percent of the patients had a characteristic anomaly: the adenohypophysis was hypoplastic, the infundibulum was absent and the posterior pituitary lobe was ectopic at the bottom of the median eminence. The height of the anterior pituitary was significantly reduced in these patients (1.9 +/- 0.1 mm; mean +/- SD) when compared to age-matched healthy controls (4.1 +/- 0.8 mm, p<0.001) or hypopituitary patients with a normal MRI (4.3 +/- 0.8 mm). MPHD was found in 62% of patients with the pituitary anomaly whereas only 27% of children with a normal MRI had MPHD (p<0.05). CONCLUSIONS: The pathogenesis of the pituitary anomaly is unknown; a disorder during embryonal development or perinatal events have been discussed as causal factors. MRI should have a prominent position in the work-up of hypopituitary children. When an anatomical malformation is visualized by MRI, the diagnostic terminology should be adapted accordingly.

Hertel exophthalmometry versus computed tomography and optical 3D imaging for the determination of the globe position in zygomatic fractures.

It has been the aim of the present study, to introduce the combination of computed tomography and optical 3D imaging to exophthalmometry and to compare the resulting data to the classic Hertel method. Twenty patients without orbital pathology and 12 patients were included in the study, who were subjected to a preoperative computed tomography. Optical 3D images of the facial surface were assessed and Hertel exophthalmometry was carried out to determine protrusion. In patients with zygomatic fractures the assessment of optical 3D images and Hertel values was repeated 5 days after surgery. Preoperative axial CT slices and postoperative optical contours through the globes were superimposed and the change in protrusion was determined. The protrusion values assessed either by CT, Hertel exophthalmometry or optical 3D imaging for patients without orbital pathology did not show any statistically significant differences between each other. For zygomatic fractures, Hertel exophthalmometry revealed more pronounced protrusion data in four of five cases of a posterolaterally dislocated lateral orbital rim and a higher degree of enophthalmos in cases without dislocation of the lateral orbital rim than it could be proved in the CT slices. The differences between optical measurements and CT data were minimal in patients with zygomatic fractures. The combination of computed tomography as baseline measurement and optical 3D imaging for the follow-up examinations reveal more realistic data in cases of zygomatic fractures than Hertel measurements and should be preferred.

[Cytokeratin tumor markers in ovarian carcinoma: tissue polypeptide specific antigen (TPS) and M3/M21]. / Zytokeratintumormarker beim Ovarialkarzinom: Tissue Polypeptide Specific Antigen (TPS) und M3/M21.

The aim of the present study was to evaluate the clinical usefulness of the cytokeratin tumor marker M3/M21 as a screening, prognostic, and monitoring marker for ovarian cancer and as a predictive marker in patients with adnexal masses. In order to determine the specificity of the M3/M21 test we investigated M3/M21 serum levels in several benign conditions. The cytokeratin tumor markers M3/M21 and Tissue Polypeptide Specific Antigen (TPS) were also investigated in the follow-up of ovarian cancer patients. We evaluated M3/M21 serum levels in 75 patients suffering from ovarian cancer FIGO stages Ia to III, using a prototype immunoradiometric assay (IRMA). Sera of patients with benign cysts, endometriosis, pelvic inflammatory disease, inflammatory bowel disease and liver cirrhosis were evaluated in 90, 10, 38, 10, and 20 cases, respectively. Furthermore, we analyzed TPS serum levels by means of IRMA during the follow-up of 40 patients suffering from ovarian cancer. With a sensitivity of 57% and a specificity of 95% M3/M21 was not suitable as a screening marker for ovarian cancer. Although M3/M21 was able to discriminate between ovarian cancer and benign adnexal tumors (univariate logistic regression, p = 0.0003), M3/M21 did not provide additional information (in addition to CA 125) (multivariate logistic regression, p = 0.2). M3/M21 serum levels were elevated in several benign conditions such as liver cirrhosis and inflammatory bowel disease. In ovarian cancer patients elevated M3/M21 serum levels prior to therapy were associated with a poor overall and disease-free survival (log-rank test, p = 0.03, and log-rank test, p = 0.01, respectively). In patients with recurrent ovarian cancer M3/M21 and TPS showed median lead-time effects of 3.2 and 3.9 months, respectively. M3/M21, while obviously not suitable for screening or differential diagnosis of adnexal masses, could be useful as an additional prognostic factor. M3/M21 and TPS are valuable tumor markers in the follow-up of ovarian cancer patients.

[Is the Gaudenz Incontinence Questionnaire still relevant in diagnosis of urodynamic function?]. / Ist der Gaudenz-Inkontinenzfragebogen in der urodynamischen Funktionsdiagnostik noch aktuell?

The results of this study show that the Gaudenz incontinence questionnaire can still be used to help determine the history of the incontinent patient. However, the questionnaire cannot substitute--Gaudenz stressed this fact in his study--a complete urogynecologic workup. Especially delicate problems as urinary incontinence require reliable and objective diagnostics and therapy using the advantages of the available diagnostic potential.

[Urethrovesical morphology in continent women in spinal anesthesia]. / Urethrovesikale Morphologie bei kontinenten Frauen in Spinalanästhesie.

OBJECTIVE: To assess the influence of spinal anesthesia on bladder neck position and a clinical stress test in continent women. METHODS: In a prospective investigation, 14 women underwent urodynamic, sonographic and clinical assessment during spinal anesthesia. Results were compared to those obtained immediately preoperatively in the same patient. RESULTS: During spinal anesthesia, the bladder neck was found to be located significantly lower and more posterior, and in 4/7 parous patients (0/7 nullipara) the clinical stress test was positive. CONCLUSION: These data provide additional evidence for the importance of neuromuscular function in the etiology of pelvic floor dysfunction and genuine stress incontinence.

Central alpha-adrenoceptors involved in cardiovascular regulation.

It appears to be established that the central alpha-adrenoceptors involved in the control of cardiovascular function belong to the alpha 2-subtype. They are stimulated by catecholamines, clonidine, and other imidazoline derivatives but also by compounds of rather heterogenous structure. Activation of these central alpha 2-adrenoceptors results in an increase of cardiac vagal tone as well as in decreases of sympathetic nervous activity, blood pressure, and heart rate. alpha 2-Adrenoceptors in the nucleus of the solitary tract appear to be the main site of action of clonidine, followed, in decreasing order of likelihood, by those in the intermediolateral cell column of the spinal cord and the anterior hypothalamus. The location of the alpha 2-adrenoceptors relative to synapses is controversial. Two possibilities have so far been discussed: a location at the terminals of adrenergic neurons (presynaptic) or on the cell bodies or dendrites of not yet identified neurons innervated by adrenergic neurons (postsynaptic). In this paper, attention is drawn to another possibility suggesting a location at the terminals of nonadrenergic neurons.

alpha-Adrenoceptor mediated contractile and electrical responses of vascular smooth muscle.

Concentration-response curves for the contractile effects of noradrenaline and the alpha 1-adrenoceptor agonist methoxamine, obtained in strips of rabbit main pulmonary artery under various experimental conditions, characterized methoxamine as a full agonist with approximately one-third of the potency of noradrenaline. As measured with intracellular glass microelectrodes, both agonists depolarized the vascular smooth muscle cells over the lower segment of the concentration-contraction curve without further depolarization over the upper segment, indicating that part of the contraction occurred in the absence of changes in membrane potential. The similarity between noradrenaline and methoxamine with regard to induction of electrical events and contraction and the complete blockade of the two types of response by the alpha 1-antagonist prazosin suggest that the effects of both agonists are mediated in the rabbit main pulmonary artery mainly, if not exclusively, by alpha 1-adrenoceptors. In contrast to the induced changes in membrane potential, membrane resistance was decreased by methoxamine over the entire range of the concentration-contraction curve as determined by measurement of the space constant of the vascular strips. This points to the occurrence of alpha 1-adrenoceptor mediated changes of membrane properties which are not reflected by alterations in membrane potential. It is concluded that contraction induced in the rabbit main pulmonary artery by stimulation of alpha 1-adrenoceptors is better described by the term electromechanical than pharmacomechanical coupling.

Modified rapid transform.

We describe a new fast, shift-invariant transform, the modified rapid transform (MRT). The MRT combines the well-known rapid transform with preprocessing steps. Computer simulations show that for 1-D binary patterns the MRT with a sufficient number of preprocessing steps may perform shift-invariant one-to-one mapping. The modification is also efficient for 2-D patterns. The MRT can be usefully applied as a preprocessing step in automatic inspection and pattern recognition, where shift invariance, uniqueness, and low computing time is required. As an example, the use of MRT in optical character recognition is discussed.

Acquisition of 3-D data by focus sensing.

A range sensing technique is demonstrated that finds the 3-D shape of diffusely reflecting objects. The technique works sequentially in depth direction and is based on structured illumination and focus sensing. A TV camera and analog electronics are used to find the locations in focus of each step of a focus series in TV real time. The depth resolution is not very high, however, the technique is simple, rapid, and well suited to get an overview of a scene in robot vision.

Testing optics by experimental ray tracing with a lateral effect photodiode.

We demonstrate a method for testing optics (spherical and aspheric) and other reflecting or transmitting objects. We call this experimental ray tracing. A laser beam is sent through the sample, and its propagation is determined with a lateral effect photodiode. A modified Hartmann test can be performed by measurement of beam location within two planes. Measurement in one plane close to the focus delivers a spot diagram. The method is well suited for testing even strong aspheric optics. As a further use we demonstrate 3-D shape measurement of nonplanar glass plates, e.g., car windshields.

Light sectioning with large depth and high resolution.

We report a method for 3-D sensing by light sectioning. The specific goal is to demonstrate that high resolution and large depth can be achieved simultaneously, thus overcoming the major limitation of conventional light sectioning. We use the diffraction pattern of an axicon to generate a light knife with large depth of field (for example, 1700 mm) and high lateral resolution (for example, 55 microm). Illuminating an object with this light knife creates a profile on the object. We detect this profile with a CCD TV camera and evaluate the centroid of the profile by means of an interpolation algorithm.

Class of transforms invariant under shift, rotation, and scaling.

We introduce a new class of nonreversible transform that is simultaneously invariant under shift, rotation, and scaling. The algorithm is based on a general transformation where the kernel itself contains the function to be transformed. Thus the invariances are achieved by a kind of self-mapping. Preprocessing of the input signal, such as determination of the centroid or coordinate transformation, is not necessary. The transform can be used for multidimensional data. Fast implementation is possible by look-up tables.

Feedback network with space invariant coupling.

Processing images by a neural network means performing a repeated sequence of operations on the images. The sequence consists of a general linear transformation and a nonlinear mapping of pixel intensities. The general (shift variant) linear transformation is time consuming for large images if done with a serial computer. A shift invariant linear transformation can be implemented much easier by fast Fourier transform or optically, but the shift invariant transform has fewer degrees of freedom because the coupling matrix is Toeplitz. We present a neural convolution network with shift invariant coupling that nevertheless exhibits autoassociative restoration of distorted images. Besides the simple implementation, the network has one more advantage: associative recall does not depend on object position.