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.

Fusion of computed tomography data and optical 3D images of the dentition for streak artefact correction in the simulation of orthognathic surgery.

OBJECTIVE: To determine the limits of accuracy of fusion of optical three-dimensional (3D) imaging and computed tomography (CT) with and without metal artefacts in an experimental setting and to show the application of this hybrid system in 3D orthognathic surgery simulation. METHODS: Ten plaster casts of dental arches were subjected to a CT scan and optical 3D surface imaging. Subsequently, the first molars in the plaster casts were supplied with metal restorations, bilaterally, and new CT scans and optical surface images were assessed. The registration of the surface data of the two imaging modalities of the study models without and with metal restorations was carried out. The mean distance between the two data sets was calculated. From a patient a CT scan of the skull as well as optical 3D images of plaster casts of the dental arches were acquired. Again the two imaging modalities were registered and virtual orthognathic surgery simulation was carried out. RESULTS: The mean distance between the corresponding data points of CT and optical 3D surface images was 0.1262+/-0.0301 mm and 0.2671+/-0.0580 mm, respectively, for the plaster casts without and with metal restorations. The differences between these data were statistically significant (P<0.0005). For the patient case a mean difference of 0.66+/-0.49 mm and 0.56+/-0.48 mm for mandible and maxilla, respectively, was calculated between CT and optical surface data. CONCLUSION: The accuracy of the fusion of 3D CT surface data and optical 3D imaging is significantly reduced by metal artefacts. However, it seems appropriate for virtual orthognathic surgery simulation, as post-operative orthodontics are performed frequently.

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.

Integrins and extracellular matrix proteins in the human childhood and adolescent growth plate.

Interaction of chondrocytes with the surrounding matrix significantly influences differentiation and growth. These processes involve cell surface proteins, particularly integrins. The aim of this study was to compare the expression of integrins (alpha1, alpha2, alpha3, alpha5, alpha6, alphav, beta1, beta3, and beta5 subunits) together with matching binding proteins in human childhood and adolescent growth plate cartilage using immunohistochemistry. Integrin beta1 was detected in all chondrocytes of the growth plate cartilage, beta3 only in osteoclasts of the opening zone, and beta5 in hypertrophic chondrocytes and osteoblasts. Integrin alpha1, alpha2, and alpha5 subunits were expressed by chondrocytes in the proliferative and hypertrophic zone as well as in osteoblasts and osteoclasts. Integrin av and alpha6 subunits were present in chondrocytes of all zones, alpha3 only in osteoclasts. Collagen type II and fibronectin were seen throughout the growth plate, collagen type X in the hypertrophic zone, collagen type I in the ossifying trabecules. Laminin was expressed by chondrocytes in the resting zone and more weakly in the proliferative zone, collagen VI was present in the pericellular and interterritorial matrix in all zones of the growth plate. These results differ from previous reports on the distribution of integrins in the fetal growth plate. However, there was no difference in integrin expression in children before and during puberty. Our results indicate that integrin expression is not influenced by endocrine factors during sexual maturation and suggest that the process of skeletal maturation is not regulated via altered integrin expression.

Three-dimensional ultrasound of the female urethra: comparing transvaginal and transrectal scanning.

OBJECTIVES: To compare transvaginal and transrectal three-dimensional ultrasound in determining the morphology and measurements of the female urethra. DESIGN: Sixty-five women who had not had surgery for incontinence or pelvic floor descent had transvaginal and transrectal sonography using a 7.5-MHz mechanical sector endoprobe with three-dimensional facilities. The multiplanar display of the scanned volumes allowed detailed morphologic assessment of the urethra and the measurement of distances and volumes. Statistical endpoints were: sagittal urethral diameter, maximum rhabdosphincter length and thickness, maximum thickness of the smooth muscle complex, and the volumes of the rhabdosphincter and the smooth muscle complex. Values were compared between the two approaches using Student's t-test and Bland-Altman analysis. RESULTS: Both vaginal and rectal scans were feasible. However, significant differences between the two approaches were found for the sagittal diameter of the urethra (8.4 +/- 1.9 mm on vaginal vs. 11.5 +/- 2.2 mm on rectal scans, P < 0.01) and the transverse diameter of the urethra's smooth muscle complex (11.2 +/- 0.3 mm on vaginal vs. 8.6 +/- 0.2 mm on rectal scans, P < 0.001). No other variables showed significant differences. Compression of the urethra and displacement under the symphysis pubis were observed when the ultrasound probe was applied vaginally. Bland-Altman analysis showed acceptable variability for differences of distances but considerable variability for the differences of volumes. CONCLUSION: The female urethra can be examined both vaginally and rectally by three-dimensional ultrasound. A transvaginally applied probe seems to have a compression effect on the urethra.

Localization and Registration of Three-Dimensional Objects in Space-Where are the Limits?

We discuss the accuracy limits for the localization of surfaces in three-dimensional (3-D) space. Such a localization is necessary for the registration of different views of an object, taken by 3-D sensors from several directions. A quantitative analysis shows that the lateral localization accuracy of a small surface area is proportional to the local curvature of the surface. This confirms the intuitive conjecture that our visual system performs localization of 3-D objects via sharp features. The longitudinal localization accuracy depends only on the noise of the data and is usually much better than the lateral localization accuracy, suggesting that surfaces are to be registered only along the longitudinal directions.

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.

Combined pituitary hormone deficiency and pituitary hypoplasia due to a mutation of the Pit-1 gene.

Several mutations of the pituitary-specific transcription factor Pit-1 have been identified. We describe a girl with a mutation of the Pit-1 gene leading to a complete lack of GH, TSH and prolactin and a marked hypoplasia of the anterior pituitary gland. The patient had a homozygous nonsense-mutation at position 172 (CGA to TGA), converting arginine into a stop codon, leading to an early termination of protein translation. During the infancy period the girl had very conspicuous symptoms of hypothyroidism and the diagnosis of thyroid insufficiency preceded the diagnosis of GH-deficiency by 1.5 years. Treatment with thyroxine and GH resulted in excellent catch-up growth.

Screening of patients with Turner syndrome for "hidden" Y-mosaicism.

The presence of Y-chromosomal sequences in the cells of patients with Turner-Syndrome (TS) is a risk factor for the development of gonadal tumors. Therefore and since demonstration of Y-material usually results in prophylactic gonadectomy optimal sensitivity and specificity of the diagnosis have to be attempted. We wanted to evaluate the diagnostic potential of cytogenetic investigations as routinely employed in TS. In the most comprehensive study published so far we screened 208 TS patients for the presence of Y-chromosomal sequences by polymerase chain reaction (PCR) specific for eight different loci along the Y-chromosome. Six patients (3%) without cytogenetic evidence of Y-chromosome were found to be Y-positive. Among 12 cases with marker chromosomes two more Y-chromosomal fragments were identified. Thus, PCR-screening for Y-specific sequences was shown to be a valuable tool in the clinical management of Turner patients.

[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.

The influence of growth hormone monotherapy and growth hormone in combination with oxandrolone or testosterone on thyroxid hormone parameters and thyroxine binding globulin in patients with Ullrich-Turner syndrome.

UNLABELLED: Administration of human growth hormone (GH) has yielded conflicting results concerning its role on thyroid function in patients with Ullrich-Turner syndrome. Therefore, we investigated the course of thyroid hormone parameters and thyroxin binding globulin in relation to GH therapy, IGF-I and additional oxandrolone-(Ox) or testosterone (T) treatment in 20 patients with Ullrich-Turner syndrome. During the 1st year the patients received only GH. There was no change in T4, fT4, and TSH levels, T3 increased significantly (P < 0.01) after 6 and 12 months, resulting in a higher T3/T4 ratio. TBG (P < 0.05) and IGF-I (P < 0.01) increased after 6 months and remained elevated at 12 months. A significant positive correlation was found between the change of T4 and TBG after 6 months (r = 0.47, P < 0.05) and after 12 months (r = 0.69, P < 0.005). Thirteen patients were further investigated after addition of an anabolic compound; 7 received Ox (0.0625 mg/kg/day po) and 6 low dose T (5 mg i.m. every 14 days). Chronological age was comparable in these groups (10.7 +/- 2.7 vs 10.7 +/- 3.6 years). After 6 months of combination therapy with Ox, T4, T3 and TSH decreased. As T4 and T3 showed a parallel decrease the T3/T4 ratio remained elevated. TBG declined after 6 and 12 months (P < 0.05), while IGF-I showed a further increment (P < 0.05). There was no correlation between the changes in T4 and IGF-I, TSH and TBG, respectively. In the T-treated group only IGF-I increased (P < 0.05) to the same extent as in the Ox-treated patients, whereas the thyroid parameters did not change. CONCLUSION: The observed changes in thyroid hormone and TBG levels in the Ox group were not mediated by GH or IGF-I. The Ox-induced TBG decrease might be linked to altered pancreatic functions regulating carbohydrate metabolism.

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.

[Imaging procedures in diagnosis of ovarian carcinoma]. / Bildgebende Verfahren in der Diagnostik des Ovarialkarzinoms.

Each of the 3 major imaging modalities, ultrasound, computed tomography, and magnetic resonance imaging, plays a specific role in the evaluation of the ovaries. Depending on the different diagnostic potential each imaging technique has its special operating areas. Ultrasound, and in particular transvaginal ultrasound, remains the examination of choice for screening and can detect a wide spectrum of ovarian pathology from simple ovarian cysts to complicated ovarian masses. Computed tomography is currently the recommended modality to stage ovarian carcinoma, but is also used in the follow-up and to judge the response to cytostatic therapy. Magnetic resonance imaging is a valuable tool to evaluate disorders of sexual differentiation and to localize ectopic gonads. Magnetic resonance imaging is also capable of depicting spread of ovarian carcinoma throughout the pelvis and the abdomen.

Observation of light propagation in volume scatterers with 10(11)-fold slow motion.

An optical method is introduced for observation of temporally and spatially resolved frames that show how light propagates in diffusely scattering materials. The method permits videos with 100-fs resolution in time to be produced. The method utilizes short-coherence interferometry. The source of information is the speckle contrast. The temporal and spatial evolution of the multiple scattering process is demonstrated for several biological and industrial samples. A major objective of the method is to investigate the conditions for optimum coherence and optimum apertures to achieve high resolution in the short-coherence interferometry. One important result is that during the propagation a sharp photon horizon evolves, which is useful for the morphological analysis of volume scatterers.

[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.

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.

[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.

Necrotizing fasciitis arising from episiotomy.

Necrotizing fasciitis is a fatal, rapidly progressive, often initially unrecognized condition. Mortality rates range from 30% to 76%. Prognosis depends on the delay of diagnosis, antimicrobial treatment and surgical excision of all necrotic tissue. A case of postpartum perineal necrotizing fasciitis arising from episiotomy is presented. Prompt recognition and aggressive therapy resulted in a favorable outcome despite significant morbidity.

Laser triangulation: fundamental uncertainty in distance measurement.

We discuss the uncertainty limit in distance sensing by laser triangulation. The uncertainty in distance measurement of laser triangulation sensors and other coherent sensors is limited by speckle noise. Speckle arises because of the coherent illumination in combination with rough surfaces. A minimum limit on the distance uncertainty is derived through speckle statistics. This uncertainty is a function of wavelength, observation aperture, and speckle contrast in the spot image. Surprisingly, it is the same distance uncertainty that we obtained from a single-photon experiment and from Heisenberg's uncertainty principle. Experiments confirm the theory. An uncertainty principle connecting lateral resolution and distance uncertainty is introduced. Design criteria for a sensor with minimum distanc uncertainty are determined: small temporal coherence, small spatial coherence, a large observation aperture.