Search

Background: A 2019 ACVIM consensus statement on diagnostics for immune-mediated hemolytic anemia (IMHA) in dogs made testing recommendations. As data on the performance of immunohematological tests was lacking, we undertook a comparative analysis., Material and Methods: Anticoagulated blood samples from 126 dogs suspected of having IMHA submitted to a diagnostic veterinary laboratory for a routine direct antiglobulin test (DAT) and from 28 healthy control dogs were evaluated for spherocytosis and autoagglutination before and after three saline washes. Samples were also subjected to different DATs: a gel minitube and an immunochromatographic strip kit used in clinics; neutral gel column cards, microtiter plates (at 4°, 22°, and 37°C), capillary tubes, and flow cytometry used in laboratories., Results: Samples from healthy dogs yielded negative results with all immunodiagnostic tests. Among the 126 samples submitted for DAT 67 were positive by a DAT utilizing microtiter plates with goat anti-dog antiglobulin DAT at 22°C. Notably, DAT results were comparable and consistent across all evaluated methods regardless of antiglobulin and temperature used. DAT+ dogs were more severely anemic and more likely to have erythroid regeneration compared to DAT- dogs. Macroscopic agglutination in tubes or on slides was observed in 48 samples after 1:1 and 1:4 blood to saline dilution, but only persisted in four samples after washing. Among the DAT+ samples, 57% had agglutination, 87% had spherocytosis, and 45% had both. There was good correlation between spherocytosis and DAT results from the six DAT techniques, but the correlation with autoagglutination was only fair. Clinical follow-up was available for 42 dogs. Of the sample from 12 DAT+ dogs collected during treatment, 10 remained DAT+ when tested 1-24 weeks after initial assessment., Conclusions: Based upon this comparative prospective survey, all in-clinic and laboratory DAT techniques produced similar results when performed by trained personnel and can therefore be recommended for detection of antibody-coated erythrocytes and immunohematological diagnosis. In addition, use of these tests for monitoring response of IMHA dogs to treatment might be valuable., (© 2021. The Author(s).)

In this study, we describe new methods for studying cancer cell metabolism with hyperpolarized 13 C magnetic resonance spectroscopy (HP 13 C MRS) that will enable quantitative studies at low oxygen concentrations. Cultured hepatocellular carcinoma cells were grown on the surfaces of non-porous microcarriers inside an NMR spectrometer. They were perfused radially from a central distributer in a modified NMR tube (bioreactor). The oxygen level of the perfusate was continuously monitored and controlled externally. Hyperpolarized substrates were injected continuously into the perfusate stream with a newly designed system that prevented oxygen and temperature perturbations in the bioreactor. Computational and experimental results demonstrated that cell mass oxygen profiles with radial flow were much more uniform than with conventional axial flow. Further, the metabolism of HP [1- 13 C]pyruvate was markedly different between the two flow configurations, demonstrating the importance of avoiding large oxygen gradients in cell perfusion experiments.

Serum feline trypsin-like immunoreactivity (fTLI) is commonly used to diagnose feline exocrine pancreatic insufficiency (EPI). This study aimed to describe signalment and clinical data of cats with EPI. Determination of TLI was performed using an in-house ELISA; the reference interval was defined using a Reference Limit Estimator. Groups were formed from 4813 cats (2019-2020), based on their fTLI concentration: 1 (<8 µg/L; decreased; n = 275), 2 (8-88 µg/L; reference interval; n = 4256), and 3 (>88 µg/L; increased; n = 282). Males and Domestic Shorthairs were most common in all groups. Group 3 had the highest (13 years), and group 1 had the lowest (9 years), median age. Clinical information was available for 200 cats (decreased fTLI: n = 87, lower reference interval (8-12 µg/L): n = 113). Treatment response was observed in 83% (decreased fTLI) and 66% (lower reference interval). EPI cats displayed weight loss (69%), diarrhoea (68%), vomiting (41%), anorexia (39%), poor hair coat (35%), lethargy (33%), and/or polyphagia (21%). The lower the serum fTLI concentration, the more often good treatment response was reported ( p = 0.022) but there were no statistically significant clinical signs. In conclusion, fTLI is a helpful parameter to diagnose EPI but predicting treatment response based on signalment or clinical signs is not possible.

This interview addresses the trajectory of harm reduction policies and practices in Brazil, in the words of harm reducer Fatima Machado. Harm reduction emerged in the 1980s in Europe, it began in Brazil in 1989 as a strategy to prevent aids among injection drug users, and then diversified and expanded. This interview focuses on the early years of developing these innovative practices and questions their current developments. Fatima Machado was one of the precursors working in the field and in activism for harm reduction in Brazil, and one of the founders of the Brazilian Association of Harm Reducers.

Background: Diagnosis of pancreatic diseases in dogs is still challenging because of variable clinical signs, which do not always correspond with clinical pathology and histopathological findings., Objectives: To characterize inflammatory and neoplastic pancreatic diseases of dogs and to correlate these findings with clinical findings and canine pancreatic lipase immunoreactivity (cPLI) results., Animals: Tissue specimens and corresponding blood samples from 72 dogs submitted for routine diagnostic testing., Methods: Four groups were defined histologically: (1) normal pancreas (n = 40), (2) mild pancreatitis (n = 8), (3) moderate or severe pancreatitis (acute, n = 11; chronic, n = 1), and (4) pancreatic neoplasms (n = 12). An in-house cPLI ELISA (<180 μg/L, normal; >310 μg/L, pancreatitis) was performed., Results: In dogs with normal pancreas, 92.5% of serum cPLI results were within the reference range and significantly lower than in dogs with mild acute pancreatitis, moderate or severe acute pancreatitis and pancreatic tumors. In dogs with moderate or severe acute pancreatitis, cPLI sensitivity was 90.9% (95% confidence interval [CI], 58.7%-99.8%). Most dogs (9/12) with pancreatic tumors (group 4) had additional pancreatic inflammation and cPLI results were increased in 10 dogs., Conclusions and Clinical Importance: High cPLI indicates serious acute pancreatitis but underlying pancreatic neoplasms should also be taken into consideration. This study confirms the relevance of histopathology in the diagnostic evaluation of pancreatic diseases., (© 2020 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

Doppler ultrasound (DUS) is frequently performed as a screening and diagnostic modality to evaluate the transjugular intrahepatic portosystemic shunt (TIPS) for short- and intermediate-term complications of the procedure. However, the clinical significance of initial frequently observed abnormalities in flow velocities, gradient, and direction in patients with covered TIPS is less studied. A retrospective study was performed between January 2005 and December 2014, and all patients undergoing covered TIPS procedure for the management of portal hypertension were included. Abnormal DUS findings were defined as intrashunt peak systolic velocities (PSVs) less than 90 or greater than 190 cm/s, intrashunt gradient greater than 50 cm/s, and abnormal flow direction in the main, right, and/or left portal veins. A total of 283 patients with adequate clinical follow-up and baseline TIPS DUS were included in the study. Median follow-up was 18.2 months. During the follow-up period, portal hypertension symptoms recurred in 83 patients who underwent TIPS angiography and/or revision. Of the 83, 57 had an elevated portosystemic gradient (>12 mm Hg) requiring angioplasty/stenting. With regard to baseline ultrasound, low PSVs were present in 88 patients (31.1%), high PSVs in 44 patients (15.5%), and elevated gradient in 98 patients (34.6%). The rate of developing TIPS stenosis in the future was not higher in patients with abnormal DUS parameters. For example, 26 (19.7%) of 132 patients with abnormal TIPS velocities developed stenosis compared with 31 (20.5%) of 151 patients with normal velocities (P = 0.9). Based on these results, abnormal DUS findings observed on baseline TIPS ultrasound examination have low predictive value for future covered TIPS dysfunction and failure.

The most common pancreatic diseases in cats are pancreatitis and exocrine pancreatic insufficiency (EPI). Non-invasive methods, such as serological quantification of feline pancreatic lipase immunoreactivity (fPLI), are often used in the diagnosis of pancreatitis. Previous studies have compared fPLI concentrations with histopathology, considered to be the gold standard for diagnosis of feline pancreatitis. However, fPLI concentrations in cats suffering from pancreatic tumours were rarely described. The aim of the present study was to determine the sensitivity and specificity of an in-house enzyme-linked immunosorbent assay (ELISA) for the quantification of fPLI in serum samples based on histopathological findings in cats diagnosed with various pancreatic diseases. Pancreatic biopsy samples from 80 cats were included. Five groups were defined on the basis of pancreatic histopathology: group 1, normal pancreas; group 2, nodular hyperplasia; group 3, mild pancreatitis; group 4, marked (moderate/severe) pancreatitis; and group 5, pancreatic neoplasia. Serum samples from all cats were tested by fPLI ELISA (<3.6 μg/l normal, 3.6-5.3 μg/l questionable, >5.3 μg/l pancreatitis). In group 1 (n = 19), serum fPLI values were within the reference interval in 74% of cases and in group 2 (n = 9) in 78%. Cats with mild pancreatitis (n = 23), marked pancreatitis (n = 11) and pancreatic neoplasms (n = 18) had significantly increased fPLI concentrations compared with group 1 (P = 0.004/0.001/≤0.0001). Cats with nodular hyperplasia had significantly lower fPLI values than cats with marked pancreatitis (P = 0.048) or tumours (P = 0.002). Serum fPLI concentrations in group 3 were <3.6 μg/l (n = 6), 3.6-5.3 μg/l (n = 4) and >5.3 μg/l (n = 13). Calculated test sensitivity for mild pancreatitis was fPLI >3.5 μg/l: 73.9% and fPLI >5.3 μg/l: 56.5%. In group 4 (n = 11), seven of nine cats (77.8%) with marked purulent pancreatitis had elevated fPLI. In group 4, a sensitivity of 81.8% was detected for fPLI >3.5 μg/l and 63.6% for fPLI >5.3 μg/l. Two cats with marked non-purulent pancreatitis had elevated fPLI, while two cats with marked purulent pancreatitis had normal fPLI values (<3.6 μg/l). In group 5, one cat with pancreatic adenoma and one with pancreatic acinar carcinoma had normal fPLI concentrations. The other cats with pancreatic adenoma (solid, n = 1; cystic, n = 4) or carcinoma (solid, n = 9; cystic, n = 2) had elevated or high fPLI values (4.1 to >40 μg/l, median 21.2 μg/l), probably caused by additional inflammation. The results of the present study confirm the importance of detailed histopathological characterization for the interpretation of clinical signs and fPLI values in feline pancreatitis. Primary pancreatic neoplasms may also lead to elevated fPLI concentrations as there is concurrent pancreatitis in most cases. However, severe pancreatic diseases, such as chronic non-purulent pancreatitis or tumours without inflammation, may result in normal fPLI values., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Purpose: To establish the capability of near-infrared fluorescence (NIRF) imaging for the detection of matrix metalloproteinase 2 (MMP-2) activity as a biomarker of vascular remodeling (VR) in arteriovenous fistulae (AVFs) in vivo., Materials and Methods: AVFs were created in the right groins of Wistar rats (n = 10), and sham procedures were performed in the contralateral groins. Fistulography via a left common carotid artery approach confirmed stenosis (> 50%) in a subset of animals (n = 5) 4 weeks after AVF creation. After administration of MMP-2-activated NIRF probe, near-infrared imaging was performed in vivo and ex vivo of both the AVF and the sham-treated vessels to measure radiant efficiency of MMP-2-activated NIRF signal over background. Histologic analyses of AVF and sham-treated vessels were performed to measure VR defined as inward growth of the vessel caused by intimal thickening., Results: AVFs demonstrated a significantly higher percentage increase in radiant efficiency over background compared with sham vessels (45.5 ± 56% vs 16.1 ± 17.8%; P = .008). VR in AVFs was associated with increased thickness of neointima staining positively for MMP-2 (161.8 ± 45.5 μm vs 73.2 ± 36.7 μm; P = .01). A significant correlation was observed between MMP-2 activity as measured by relative increase in radiant efficiency for AVFs and thickness of neointima staining positively for MMP-2 (P = .039)., Conclusions: NIRF imaging can detect increased MMP activity in remodeled AVFs compared with contralateral sham vessels. MMP-2-activated NIRF signal correlates with the severity of intimal thickening. These findings suggest NIRF imaging of MMP-2 may be used as a biomarker of the vascular remodeling underlying stenosis., (Copyright © 2018 SIR. Published by Elsevier Inc. All rights reserved.)

In adult cattle Fasciola hepatica infection usually follows a chronic subclinical course, and reduces both the milk yield and milk quality, resulting in considerable financial losses. Effective control of the disease is based on reliable identification of asymptomatically infected individuals, which now requires special parasitological or serological diagnostic tests. It is also known that F. hepatica infection induces alterations in some serum biochemical parameters. Therefore, the study was conducted to develop a model based on serum biochemical parameters allowing to identify cows at higher risk of chronic fascioliasis. Six hundred sixty eight adult dairy cows from 97 herds located in central and northeastern Poland were clinically examined, and blood and fecal samples from them were collected for a routine monitoring of fascioliasis and metabolic profile. Using the combination of fecal sedimentation test and indirect ELISA based on excretory/secretory products 203 cows, apparently healthy in clinical examination, were definitively classed as affected by (47 cows, 23.2%) or free from (156 cows, 76.8%) chronic fascioliasis. Their serum was screened for the activity of 4 enzymes (γ-glutamyl transpeptidase, glutamate dehydrogenase, aspartate aminotransferase, creatine kinase) and concentration of 18 other components (total bilirubin, total protein, albumin, globulin, urea, glucose, non-esterified fatty acids, β-hydroxybutyrate, sodium, potassium, chloride, calcium, magnesium, copper, zinc, iron, selenium, and haptoglobin). Logistic regression analysis was used to build 4 multivariable models allowing for identification of cows at risk of chronic fascioliasis. Then, the accuracy was compared between the models using the area under the receiver operating characteristic curve (AUROC), and an optimal cut-off value was determined for the most accurate model using Youden J index. The most accurate proved to be the model based on glutamate dehydrogenase activity and globulin, urea (all three positively linked with risk of chronic fascioliasis), and selenium concentration (negatively linked) adjusted by the access to pasture and cow's age. At the optimal cut-off of 0.37 this model had sensitivity of 85.1% (CI 95%:72.3%, 92.6%), specificity of 90.4% (CI 95%:84.7%, 94.1%), positive likelihood ratio of 8.8 (CI 95%:5.4, 14.5), and negative likelihood ratio of 0.16 (CI 95%:0.08, 0.33). This model performed significantly better than model including only γ-glutamyl transpeptidase or model including both γ-glutamyl transpeptidase and aspartate aminotransferase (both also adjusted by the access to pasture and cow's age), and was the only model which performed significantly better than the basic model based solely on the access to pasture and cow's age., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Fasciola hepatica and Ostertagia ostertagi infections are widespread in cattle population of Europe, however data on their prevalence in Poland are only fragmentary. Therefore, the cross-sectional study was carried out to determine the herd-level seroprevalence of F. hepatica and O. ostertagi infection in dairy cattle population in the central and north-eastern provinces Poland, and to identify basic local risk factors for these infections. In total, 598 herds were enrolled, 394 (65.9%) in the north-eastern province and 204 (34.1%) in the central province. In each herd the questionnaire survey was conducted and bulk-tank milk (BTM) sample was collected and screened using two indirect immunoenzymatic tests. Optical density ratio (ODR) was regarded as the quantitative proxy of exposure to either of the two parasites., Results: Both Fasciola and Ostertagia ELISA ODR in the north-eastern province was significantly higher than ODR in the central province. At the cut-off value of ODR = 0.27 the true herd-level seroprevalence of F. hepatica was 79.6% (95% CI: 74.0%, 84.3%) in the north-eastern province and 13.0% (95% CI: 5.3%, 21.7%) in the central province. At the cut-off of ODR = 0.50151 of 188 herds (80.3%, 95% CI: 74.1%, 85.4%) were seropositive for O. ostertagi in the north-eastern province and only 70 of 136 herds (51.5%, 95% CI: 43.1%, 59.7%) were seropositive in the central province. Location of a herd in the north-eastern province, longer grazing period practiced in a herd and > 50%-share of grazing grass in roughage were all positively related to the increase in exposure to both parasites. Moreover, the use of hay or haylage as main roughage proved to be positively related to the increase in exposure to F. hepatica., Conclusions: F. hepatica and O. ostertagi are widespread in cattle population in Poland, however their occurrence at a herd-level varies between different regions of Poland. This diversity can only partly be explained by different herd management, and appears linked to environmental and climate conditions typical for these regions.

Purpose: To explore quantitative differences between genders in morphologic colonic metrics and determine metric reproducibility., Methods: Quantitative colonic metrics from 20 male and 20 female CTC datasets were evaluated twice by two readers; all exams were performed after incomplete optical colonoscopy. Intra-/inter-reader reliability was measured with intraclass correlation coefficient (ICC) and concordance correlation coefficient (CCC)., Results: Women had overall decreased colonic volume, increased tortuosity and compactness and lower sigmoid apex height on CTC compared to men (p<0.0001,all). Quantitative measurements in colonic metrics were highly reproducible (ICC=0.9989 and 0.9970; CCC=0.9945)., Conclusion: Quantitative morphologic differences between genders can be reproducibility measured., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Purpose To characterize hepatocellular carcinoma (HCC) cells surviving ischemia with respect to cell cycle kinetics, chemosensitivity, and molecular dependencies that may be exploited to potentiate treatment with transarterial embolization (TAE). Materials and Methods Animal studies were performed according to institutionally approved protocols. The growth kinetics of HCC cells were studied in standard and ischemic conditions. Viability and cell cycle kinetics were measured by using flow cytometry. Cytotoxicity profiling was performed by using a colorimetric cell proliferation assay. Analyses of the Cancer Genome Atlas HCC RNA-sequencing data were performed by using Ingenuity Pathway Analysis software. Activation of molecular mediators of autophagy was measured with Western blot analysis and fluorescence microscopy. In vivo TAE was performed in a rat model of HCC with (n = 5) and without (n = 5) the autophagy inhibitor Lys05. Statistical analyses were performed by using GraphPad software. Results HCC cells survived ischemia with an up to 43% increase in the fraction of quiescent cells as compared with cells grown in standard conditions (P < .004). Neither doxorubicin nor mitomycin C potentiated the cytotoxic effects of ischemia. Gene-set analysis revealed an increase in mRNA expression of the mediators of autophagy (eg, CDKN2A, PPP2R2C, and TRAF2) in HCC as compared with normal liver. Cells surviving ischemia were autophagy dependent. Combination therapy coupling autophagy inhibition and TAE in a rat model of HCC resulted in a 21% increase in tumor necrosis compared with TAE alone (P = .044). Conclusion Ischemia induces quiescence in surviving HCC cells, resulting in a dependence on autophagy, providing a potential therapeutic target for combination therapy with TAE. © RSNA, 2017 Online supplemental material is available for this article.

Aim: To assess for passive expansion of sub-maximally dilated transjugular intrahepatic portosystemic shunts (TIPS) and compare outcomes with maximally dilated TIPS., Methods: Polytetrafluoroethylene covered TIPS (Viatorr) from July 2002 to December 2013 were retrospectively reviewed at two hospitals in a single institution. Two hundred and thirty patients had TIPS maximally dilated to 10 mm (mTIPS), while 43 patients who were at increased risk for hepatic encephalopathy (HE), based on clinical evaluation or low pre-TIPS portosystemic gradient (PSG), had 10 mm TIPS sub-maximally dilated to 8 mm (smTIPS). Group characteristics (age, gender, Model for End-Stage Liver Disease score, post-TIPS PSG and clinical outcomes were compared between groups, including clinical success (ascites or varices), primary patency, primary assisted patency, and severe post-TIPS HE. A subset of fourteen patients with smTIPS underwent follow-up computed tomography imaging after TIPS creation, and were grouped based on time of imaging (< 6 mo and > 6 mo). Change in diameter and cross-sectional area were measured with 3D imaging software to evaluate for passive expansion., Results: Patient characteristics were similar between the smTIPS and mTIPS groups, except for pre-TIPS portosystemic gradient, which was lower in the smTIPS group (19.4 mmHg ± 6.8 vs 22.4 mmHg ± 7.1, P = 0.01). Primary patency and primary assisted patency between smTIPS and mTIPS was not significantly different ( P = 0.64 and 0.55, respectively). Four of the 55 patients (7%) with smTIPS required TIPS reduction for severe refractory HE, while this occurred in 6 of the 218 patients (3%) with mTIPS ( P = 0.12). For the 14 patients with follow-up computed tomography (CT) imaging, the median imaging follow-up was 373 d. There was an increase in median TIPS diameter, median percent diameter change, median area, and median percent area change in patients with CT follow-up greater than 6 mo after TIPS placement compared to follow-up within 6 mo (8.45 mm, 5.58%, 56.04 mm 2 , and 11.48%, respectively, P = 0.01)., Conclusion: Passive expansion of smTIPS does occur but clinical outcomes of smTIPS and mTIPS were similar. Sub-maximal dilation can prevent complications related to over-shunting in select patients., Competing Interests: Conflict-of-interest statement: None of the authors has any conflict of interest to report.

The aim of this study was to evaluate feasibility and reproducibility of quantitative assessment of colonic morphology on CT colonography (CTC). CTC datasets from 60 patients with optimal colonic distension were assessed using prototype software. Metrics potentially associated with poor endoscopic performance were calculated for the total colon and each segment including: length, volume, tortuosity (number of high curvature points <90°), and compactness (volume of box containing centerline divided by centerline length). Sigmoid apex height relative to the lumbosacral junction was also measured. Datasets were quantified twice each, and intra-reader reliability was evaluated using concordance correlation coefficient and Bland-Altman plot. Complete quantitative datasets including the five proposed metrics were generated from 58 of 60 (97 %) CTC examinations. The sigmoid and transverse segments were the longest (55.9 and 51.4 cm), had the largest volumes (0.410 and 0.609 L), and were the most tortuous (3.39 and 2.75 high curvature points) and least compact (3347 and 3595 mm 2 ), noting high inter-patient variability for all metrics. Mean height of the sigmoid apex was 6.7 cm, also with high inter-patient variability (SD 6.8 cm). Intra-reader reliability was high for total and segmental lengths and sigmoid apex height (CCC = 0.9991) with excellent repeatability coefficient (CR = 3.0-3.3). There was low percent variance of metrics dependent upon length (median 5 %). Detailed automated quantitative assessment of colonic morphology on routine CTC datasets is feasible and reproducible, requiring minimal reader interaction.

Purpose: To characterize thermal changes induced by rheolytic thrombectomy (RT) within an ex vivo venous model and evaluate resultant changes of endothelial and vessel wall injury., Materials and Methods: Patent human saphenous vein segments without thrombus were mounted in an ex vivo perfusion system with a temperature probe apposed to the adventitial surface. RT was performed over a guide wire to facilitate device centering. Continuous RT was performed for 4 minutes with temperature recorded every 10 seconds. Pulsed RT was performed for eight cycles of 30 seconds followed by 10 seconds of deactivation. Mean temperature increase, maximum temperature (Tmax), intimal/medial thickness, endothelial cell staining (CD31), and heat shock protein 90 (HSP90) expression were compared between untreated and RT-treated venous segments., Results: Continuous RT produced a mean 7.6°C increase in temperature above baseline with mean Tmax of 44.1°C. Pulsed RT produced a mean 7.3°C increase in temperature and mean Tmax of 43.8°C. Differences in mean temperature increase (P = .66) and Tmax (P = .71) between the two groups were not statistically significant. RT-treated segments showed intima/media thinning (0.32 mm before RT and 0.18 mm after RT; P = .004) and reduction in intact endothelium (38.8% before RT and 13.8% after RT; P = .002). Staining for HSP90 showed a 3.1% increase in expression after RT (P = .31)., Conclusions: RT in this venous model showed reproducible increases in vessel temperature and evidence of endothelial and vessel wall injury. Avoiding prolonged RT application to a focal vascular segment during clinical use may be beneficial., (Copyright © 2016 SIR. Published by Elsevier Inc. All rights reserved.)

In recent decades, there have been numerous advances in the management of liver cancer, cirrhosis, and diabetes mellitus. Although these diseases are wide ranging in their clinical manifestations, each can potentially be treated by exploiting the blood flow dynamics within the portal venous system, and in some cases, adding cellular therapies. To aid in the management of these disease states, minimally invasive transcatheter portal venous interventions have been developed to improve the safety of major hepatic resection, to reduce the untoward effects of sequelae from end-stage liver disease, and to minimize the requirement of exogenously administered insulin for patients with diabetes mellitus. This state of the art review therefore provides an overview of the most recent data and strategies for utilization of preoperative portal vein embolization, transjugular intrahepatic portosystemic shunt placement, balloon retrograde transvenous obliteration, and islet cell transplantation., (© RSNA, 2016.)

Objectives: Assess differences in three-dimensional colonic metrics on CTC in women with or without hysterectomy following incomplete endoscopy to determine if there is a correlation between colonic morphology and incomplete colonoscopy after hysterectomy., Methods: Quantitative rectosigmoid metrics were derived from CTC datasets of 37 women with hysterectomy and 36 women without hysterectomy who underwent CTC for incomplete endoscopy. Evaluated metrics included colonic length, volume, tortuosity, and compactness and sigmoid apex height relative to the lumbosacral junction. Differences were measured using the Student's t test, and intra-reader reliability was assessed using ICC. The relative risk of incomplete rectosigmoid visualization was determined by reviewing the endoscopy reports., Results: Women with hysterectomy had a lower sigmoid apex height (p = 0.002), as well as increased tortuosity (p = 0.012) and compactness (p = 0.001) and decreased length (p = 0.026) and volume (p = 0.016) of the rectosigmoid. Intra-reader reliability was high for centerline length (ICC = 0.9940) and sigmoid apex height (ICC = 0.9851). The relative risk of incomplete visualization of the rectosigmoid on endoscopy in women with hysterectomy was 2.068 (p = 0.043) compared to women without hysterectomy., Conclusion: Our pilot data show reproducible quantitative differences in three-dimensional metrics of the rectosigmoid in women with or without hysterectomy who underwent CTC for incomplete endoscopy and increased relative risk of incomplete endoscopic visualization of the rectosigmoid after hysterectomy. Our findings suggest that women with hysterectomy may benefit from CTC rather than endoscopy as the initial diagnostic test for evaluating the colon.

A serosurvey was carried out to estimate the herd-level seroprevalence of Neospora caninum infection in cattle in central and northeastern Poland. Ninety seven dairy cattle herds from 2 provinces of Poland (Podlaskie, 47 herds and Łodzkie, 50 herds) were randomly enrolled in the study using two-stage cluster method. A simple random selection was applied within each herd to select a sample of adult cows (≥18 month-old). A total number of 734 cows were enrolled in the study. The animals were screened with a commercial competitive ELISA (Bio-X Diagnostics, Belgium). To calculate true herd-level seroprevalence test sensitivity and specificity were adjusted from an individual- to a herd-level using FreeCalc method. The true overall herd-level seroprevalence of N. caninum infection was 56.7% (95% CI: 47.5%, 65.9%). The true herd-level seroprevalence in Podlaskie was 63.3% (95% CI: 43.0%, 83.6%) and 50.5% (95% CI: 32.8%, 68.2%) in Łodzkie province and these figures did not differ significantly between the two provinces (chi2 test p = 0.238). One hundred forty three of 734 cows (19.5%) were seropositive which gave the true overall individual-level seroprevalence of 20.1% (95% CI: 17.4%, 23.2%). Percentage of seropositive cows in each herd varied from 6% to 80%. This study is the first epidemiological investigation of herd-level seroprevalence of N. caninum infection in Polish dairy cattle population. In conclusion, the result of the study confirmed previous data that N. caninum infection is widespread in the Polish cattle population and thus should be considered as a potential cause of spontaneous abortions.

Purpose: To evaluate long-term patency and symptomatic recurrence rates following transjugular intrahepatic portosystemic shunt (TIPS) creation with expanded polytetrafluoroethylene (ePTFE)-covered stent grafts and to determine the necessity of extended clinical follow-up beyond 2 years after TIPS creation., Materials and Methods: A retrospective review including 262 TIPSs created with ePTFE-covered stent grafts between July 2002 and October 2012 was performed. Primary, primary assisted, and secondary patency rates were calculated. Assessment of clinical data included technical, hemodynamic, and clinical success rates, as well as mortality after TIPS creation., Results: Primary patency rates at 2, 4, and 6 years were 74%, 62%, and 50%, respectively. Primary assisted patency rates at 2, 4, and 6 years were 93%, 85%, and 78%, respectively. Secondary patency rates at 2, 4, and 6 years were 99%, 91%, and 84%, respectively. Technical and hemodynamic success rates were 99% and 93%, respectively. Clinical success rates for refractory ascites were 66% (complete response) and 90% (partial response); clinical success rate for bleeding/varices was 90%. Mortality rates at 2, 4, and 6 years after TIPS creation were 27%, 38%, and 46%, respectively. At the median wait time until transplantation, patients had an 84% chance of being alive. TIPS dysfunction developed in 21% of patients; 30% of revisions occurred later than 2 years during follow-up., Conclusions: Beyond 2 years after TIPS creation, patency rates gradually decrease, mortality rates continue to increase, and the chance of recurrent ascites or bleeding remains present. Together, these findings suggest that continued clinical follow-up beyond 2 years is necessary in patients with a TIPS created with an ePTFE-covered stent graft., (Copyright © 2015 SIR. Published by Elsevier Inc. All rights reserved.)

Purpose: To evaluate the technical feasibility of a coaxial electrode configuration to rapidly create a mechanically defined electrochemical ablation zone monitored by magnetic resonance (MR) imaging in real time., Materials and Methods: A direct current generator supplied the nitinol cathode cage and central platinum anode for coaxial electrochemical ablation. Safety and efficacy were evaluated by measuring local pH, temperature, and current scatter in saline solutions. Ablation zone diameters of 3-6 cm (n = 72) were created on ex vivo bovine liver and verified by gross pathology. Feasibility of MR monitoring was evaluated using 8 swine livers to create ablations of 3 cm (n = 12), 4 cm (n = 4), and 5 cm (n = 4) verified by histology., Results: Local pH was 3.2 at the anode and 13.8 at the cathode. Current scatter was negligible. Ablation progress increased relative to local ion concentration, and MR signal changes corresponded to histologic findings. In the ex vivo model, the times to achieve complete ablation were 15 minutes, 20 minutes, 35 minutes, and 40 minutes for diameters of 3 cm, 4 cm, 5 cm, and 6 cm, respectively. Ablation times for the in situ model were 15 minutes, 35 minutes, and 50 minutes for 3 cm, 4 cm, and 5 cm, respectively., Conclusions: The coaxial configuration mechanically defined the electrochemical ablation zone with times similar to comparably sized thermal ablations. MR compatibility allowed for real-time monitoring of ablation progress., (Copyright © 2015 SIR. Published by Elsevier Inc. All rights reserved.)

Background: Sting challenge with a live insect remains the best test for proving the efficacy of immunotherapy in Hymenoptera allergy., Objective: We studied the impact of tolerated sting challenge on quality of life., Patients and Methods: In this prospective study, data were collected via self-report questionnaires completed by consenting patients with Hymenoptera venom allergy on venom immunotherapy before and after a sting challenge., Results: The study population comprised 100 adult patients (82 with yellow jacket allergy and 18 with honeybee allergy) who participated between September 2009 and November 2010. After the sting challenge, the score on the Vespid Allergy Quality of Life Questionnaire revealed a statistically significant improvement (mean [SD] change, 0.73 [0.98]; P < .0001; 95% CI, 0.52-0.94). This improvement was independent of the patients' gender and age and the severity of the initial anaphylactic reaction. A statistically significant improvement was documented in 2 subgroups of the Short Form 36 Health Survey (physical functioning, mean change, -5.78 [25.23]; P = .038; 95% CI, -11.22 to -0.34; vitality, mean change -4.29 [12.49]; P =.002; 95% CI, -7.02 to -1.57)., Conclusions: Sting challenge results in a significant improvement in disease-specific quality of life and subgroups of general quality of life in patients allergic to Hymenoptera venom receiving established venom immunotherapy.

Background: The use of selective cyclooxygenase (COX) 2 inhibitors as an alternative to aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDs) has been suggested for patients with aspirin-exacerbated respiratory disease (AERD)., Objective: To evaluate tolerability to etoricoxib, a second-generation COX-2 inhibitor with high in vitro selectivity for COX-2 in patients with AERD., Methods: We conducted a retrospective review of patients with suspected aspirin intolerance seen between October 2007 and April 2012. Single-blind, placebo-controlled oral challenges with increasing doses of aspirin and etoricoxib were performed on 3 different days., Results: Of 262 patients with suspected aspirin intolerance, 248 underwent challenge testing with aspirin and 122 (49.2%) showed positive test results. In 104 of these aspirin-sensitive patients, etoricoxib was tested as an alternative drug and was tolerated in all but 3 (2.9%), who developed a positive asthmatic reaction., Conclusions: The highly selective COX-2 inhibitor etoricoxib was tolerated in most but not all patients tested. An oral provocation test is therefore recommended before prescribing etoricoxib for patients with AERD.

Blood was obtained from 61 neonatal Holstein calves originating from a farm in Germany with a high incidence of bovine neonatal pancytopenia (BNP). In order to detect alterations that might be related to BNP, selected haematological analytes were determined. Haematological examinations demonstrated alterations in at least two of the three cell lineages in 10 calves (16.39 per cent). Six animals (9.84 per cent) developed a bleeding disorder indicative of BNP at approximately two weeks of age. None of these animals showed alterations in complete blood cell count at sampling in the first week of life. In weeks when calves with BNP were born, an increase in the number of apparently healthy calves demonstrating decreases in blood cell counts was observed.

The hair follicle is one of the skin appendages that develops through reciprocal epithelial-mesenchymal interactions. Although a large number of studies have been made on the mechanisms of hair follicle development, the whole molecular mechanism that governs hair follicle development remains poorly defined. To further understand the molecular basis of hair follicle development, it is necessary to identify genes that drive hair morphogenesis. As an initial approach, we attempted to identify gene products associated with mouse hair follicle development. Genes upregulated in the vibrissal hair placodes were screened by polymerase chain reaction (PCR)-based cDNA subtraction. The genes thus isolated were evaluated for their hair development-associated induction and spatiotemporal expression by quantitative reverse-transcription-PCR analysis and whole-mount in situ hybridization, respectively. Finally, we identified four genes whose upregulation and spatiotemporal expression in developing hair follicles were confirmed. Successful identification of novel hair development-associated genes will be informative as clues for further characterization of hair follicle development at the molecular level., (© 2010 Japanese Dermatological Association.)

Transplantation of cryopreserved heart valves (allografts) is limited by immune responses, inflammation, subsequent structural deterioration and an expensive infrastructure. In previous studies we demonstrated that conventional frozen cryopreservation (FC) is accompanied by serious alterations of extracellular matrix (ECM) structures. As the main culprit of the observed damages ice crystal formation was identified. Objective of this study was the application principles of cryoprotection as observed in nature, occurring in animals or plants, for ice-free cryopreservation (IFC) of heart valves. Using IFC, valves were processed and stored above the glass transition temperature of the cryoprotectant formulation (-124 degrees C) at -80 degrees C to avoid any ice formation, tissue-glass cracking and preserving ECM. After implantation in the orthotopic pulmonary position in sheep, we demonstrate that IFC resulted in cell free matrices, while maintaining crucial ECM-components such as elastin and collagen, translating into superior hemodynamics. In contrast, we reveal that FC valves showed ECM damage that was not restored in vivo, and T-cell inflammation of the stroma with significant leaflet thickening. Compared to currently applied FC practice IFC also reduced infrastructural needs for preservation, storage and shipping. These results have important implications for clinical valve transplantation including the promise of better long-term function and lower costs., (Copyright 2010 Elsevier Ltd. All rights reserved.)

Basement membranes (BMs) have been implicated in cell fate determination during development. Embryoid bodies (EBs) derived from mouse embryonic stem cells deficient in the laminin gamma1 chain are incapable of depositing a BM, resulting in failure of primitive ectoderm epithelialization. To elucidate the mechanisms involved in this phenomenon, we compared the gene expression profiles of EBs with or without a BM to identify the genes showing BM-dependent expression. We found that the expressions of marker genes for the epithelial-mesenchymal transition (EMT), including the transcription factor Snai2, were up-regulated in LAMC1(-/-) EBs, whereas restoration of a BM to LAMC1(-/-) EBs suppressed the up-regulation of these genes. Overexpression of Snai2 induced the EMT in control EBs by molecular and morphological criteria, suggesting that suppression of the EMT regulatory genes is involved in BM-dependent epithelialization of primitive ectoderm. Despite the failure of primitive ectoderm epithelialization in BM-deficient EBs, mesodermal differentiation was not compromised, but rather accelerated. Furthermore, at later stages of control EB differentiation, the BM was disrupted at the gastrulation site where mesodermal markers were strongly expressed only in cells that had lost contact with the BM. Taken together, these results indicate that the BM prevents the EMT and precocious differentiation of primitive ectoderm toward mesoderm in EBs, implying that BMs are important for the control of mammalian gastrulation.

The proteasome inhibitor bortezomib was tested in a cell screen as a single agent with good efficacy in multiple hematologic and solid cancer cell lines. Phase II/III studies have supported the use of bortezomib in hematologic malignancies. In solid tumors, however, the results have been poor. There is data that bortezomib can induce PTEN expression resulting in down-regulation of PI3K-Akt signaling. We and others have shown that down-regulation of Akt results in radiation sensitization. We therefore evaluated the use of bortezomib in the head and neck cancer cell line SQ20B as a radiation sensitizer. SQ20B have a constitutively active mutation in EGFR resulting in a robust Akt response. We found that 10 nM of bortezomib decreased Akt signaling to almost undetectable. This same concentration decreased the surviving fraction after 2 Gy (SF2) from 0.77 to 0.45. Given that radiation is usually given at 2 Gy increments daily for 30 or more treatments, the exponential difference in log kill could be as high as 7 logs. The dose of bortezomib is also 2 logs less as a sensitizer than that required for single agent efficacy. Further studies should be done to explore this model in vivo.

Parietal endoderm-like cells, including Engelbreth-Holm-Swarm tumor and differentiated F9 embryonal carcinoma cells, produce huge amounts of basement membrane components, including laminin-1 (alpha1beta1gamma1). We employed a double-lox system-based gene-swapping strategy in F9 cells to replace the laminin alpha1 gene with a laminin alpha5 minigene. The gene-swapped F9 cells secreted laminin-10 (alpha5beta1gamma1) consisting of the exogenous alpha5 subunit and endogenous beta1 and gamma1 subunits on differentiation. The laminin-10 concentration in the conditioned medium exceeded 10 mg/l, which is 10-fold higher than the concentrations achieved by conventional recombinant expression systems. The gene-swapped F9 cells deposited basement membrane-like matrices containing laminin-10 on culture dishes, offering a novel microenvironment for in vitro cell manipulation.

Mycobacterium avium paratuberculosis (M.a.p.) resides and replicates in macrophages. Many of the of immune mechanisms aiding M.a.p. survival in the host's cells are known. However, little is known about interactions of M.a.p. with dendritic cells (DC). As DC are important for the induction of protective immunity against infectious diseases, we investigated the interaction of M.a.p. with these cells. Quantitative real-time PCR (RT-PCR) was used to analyse differential expression of cytokine genes after 6 h and 24 h of incubation by immature DC that phagocytosed either M.a.p. or Escherichia coli (E. coli). We hypothesized that phagocytosis of E. coli would induce pro-inflammatory cytokines due to abundant presence of lipopolysaccharide (LPS) and that the cytokine expression profile induced by phagocytosis of live M.a.p. would differ. In addition we hypothesized that incubation of immature DC with rHsp70, an immunodominant antigen of M.a.p., would induce a similar profile of cytokine gene expression as phagocytosis of intact M.a.p. However, phagocytosis of both E. coli and M.a.p. resulted in a cytokine gene expression pattern representative of a (pro-)inflammatory reaction, dominated by strong induction of IL-12 gene expression, that was higher after 24 h than after 6 h of incubation, although the response to M.a.p. was less vigorous than to E. coli. Incubation with rHsp70 resulted in a more inhibitory type of cytokine gene expression, with delayed IL-12 gene expression and downregulation of the genes for IL-1beta and IL-6 after 24 h of incubation. We conclude that bovine DC produce an immuno-stimulatory, anti-mycobacterial response to infection with M.a.p., while Hsp70 potentially contributes to pathogen virulence by allowing the bacteria to invade the host cell.

We used PCR-based cDNA subtraction to screen for genes up-regulated during mouse hair morphogenesis. One gene selected was predominantly expressed at the tip of developing hair follicles and encoded a protein characterized by the presence of twelve tandem repeats of approximately 120 amino acids and a novel N-terminal domain containing an Arg-Gly-Asp cell-adhesive motif. Immunohistochemistry demonstrated that the protein encoded by this gene, named QBRICK, was localized at the basement membrane zone of embryonic epidermis and hair follicles, in which it was more enriched at the tip rather than the stalk region. Cell adhesion assays showed that QBRICK was active in mediating cell-substratum adhesion through integrins containing alphav or alpha8 chain, but not integrin alpha5beta1. Immunohistochemistry showed that QBRICK colocalized with alphav-containing integrins in the interfollicular region, but with the alpha8-containing integrin at the tip region of developing hair follicles. These results, together, indicate that QBRICK is an adhesive ligand of basement membrane distinctively recognized by cells in the embryonic skin and hair follicles through different types of integrins directed to the Arg-Gly-Asp motif.

We screened for genes specifically expressed in the mesenchymes of developing hair follicles using representational differential analysis; one gene identified was MAEG, which encodes a protein consisting of five EGF-like repeats, a linker segment containing a cell-adhesive Arg-Gly-Asp (RGD) motif, and a MAM domain. Immunohistochemistry showed that MAEG protein was localized at the basement membrane of embryonic skin and developing hair follicles, while MAEG expression diminished at the tip of the hair bud. A recombinant MAEG fragment containing the RGD motif was active in mediating adhesion of keratinocytes to the substratum in an RGD-dependent manner. One of the adhesion receptors recognizing the RGD motif was found to be the alpha8beta1 integrin, the expression of which was detected in the placode close to MAEG-positive mesenchymal cells, but later became restricted to the tip of the developing hair bud. Given its localized expression at the basement membrane in developing hair follicles and the RGD-dependent cell-adhesive activity, MAEG may play a role as a mediator regulating epithelial-mesenchymal interaction through binding to RGD-binding integrins including alpha8beta1 during hair follicle development.

During early rodent development, the parietal endoderm appears from an inner cell mass and produces large amounts of basement membrane components, such as laminin-1 and collagen IV. To elucidate the regulatory network for gene expression during these procedures, we constructed a series of short interfering RNA expression vectors targeted to various transcription factors, transfected them into F9 embryonal carcinoma cells, and evaluated the effects of the gene silencing on the induction of parietal endoderm differentiation and basement membrane component production by treating F9 cells with all trans-retinoic acid and dibutyryl cyclic AMP. Among the transcription factors tested, silencing of Sox7 or combined silencing of Gata-4 and Gata-6 resulted in suppression of cell shape changes and laminin-1 production, which are the hallmarks of parietal endoderm differentiation. In cells silenced for Sox7, induction of Gata-4 and Gata-6 by retinoic acid and cyclic AMP treatment was inhibited, while induction of Sox7 was not affected in cells silenced for Gata-4 and Gata-6, indicating that Sox7 is an upstream regulatory factor for these Gata factors. Nevertheless, silencing of Sox7 did not totally cancel the action of retinoic acid, since upregulation of coup-tf2, keratin 19, and retinoic acid receptor beta2 was not abolished in Sox7-silenced F9 cells. Although overexpression of Sox7 alone was insufficient to induce parietal endoderm differentiation, overexpression of Gata-4 or Gata-6 in Sox7-silenced F9 cells restored the differentiation into parietal endoderm. Sox7 is therefore required for the induction of Gata-4 and Gata-6, and the interplay among these transcription factors plays a crucial role in parietal endoderm differentiation.

Purpose: Locoregional recurrence is the dominant form of treatment failure in head and neck (H&N) cancer. The epidermal growth factor receptor (EGFR) is frequently amplified in this disease (