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Exosomes are a specific subpopulation of extracellular vesicles that are widely released by cells of different origins with divergent functions that make their way into body fluids that can be conveniently sampled. In the current study, we isolated and evaluated exosomes from concurrently collected samples of milk, plasma, saliva, and urine from a group of 6 pregnant Holstein-Friesian dairy cows (aged 7 mo, 174 to 203 d of gestation). The cows had BCS of 3.5 to 5.25 (on a scale of 1 to 10), and the milk production for the season to the time of sampling ranged between 5,118 and 6,959 kg. The low levels of extracellular vesicles in saliva and urine (more than 86% fewer compared to the extracellular vesicles in milk and plasma) precluded further detailed evaluation since utility for diagnostics was deemed unlikely. In exosomes isolated from milk and plasma, size distribution, morphology, and the presence of exosome markers was confirmed by nanoparticle tracking analysis, electron microscopy, and Western blot. In addition, a targeted proteomic approach using the quadrupole ion trap mass spectrometer was also used in the study to screen for the exosome marker (e.g., Tumor susceptibility gene 101). Following confirmation of the presence of exosomes, the proteomic profiles of milk and plasma exosomes were evaluated using information-dependent acquisition-mediated liquid chromatography-tandem mass spectrometry (LC-MS/MS). The milk exosomes contain proteins that differed greatly from the plasma exosomes, with only 8 similar proteins harbored in both the milk and plasma exosomes. The milk and plasma exosomes were found to contain proteins (e.g., immunoglobulin J chain and α2 macroglobulin) associated with specific biological processes and molecular functions. Hence, the fluid of origin required for exosome analysis will be dependent on the specific information needed. In conclusion, isolated exosomes from milk and plasma samples collected at the same time point from the same dairy cows encapsulated different profiles of proteins associated with different biological processes and molecular functions.

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The greatest risk of metabolic and infectious disease in dairy cows is during the transition from pregnancy to lactating (i.e., the transition period). The objective of this experiment was to determine the effects of extracellular vesicles (microvesicles involved in cell-to-cell signaling) isolated from transition cows on target cell function. We previously identified differences in the protein profiles of exosomes isolated from cows divergent in metabolic health status. Therefore, we hypothesized that these exosomes would affect target tissues differently. To investigate this, 2 groups of cows (n = 5/group) were selected based on the concentration of ß-hydroxybutyrate and fatty acids in plasma and triacylglycerol concentration in liver at wk 1 and 2 postcalving. Cows with high concentrations of ß-hydroxybutyrate, fatty acids, and triacylglycerol were considered at increased risk of clinical disease during the transition period (high-risk group; n = 5) and were compared with cows that had low concentrations of the selected health indicators (low-risk group; n = 5). At 2 time points during the transition period (postcalving at wk 1 and 4), blood was sampled and plasma exosomes were isolated from the high-risk and low-risk cows. The exosomes were applied at concentrations of 10 and 1 µg/mL to 5 × 103 Madin-Darby bovine kidney cells grown to 50% confluence in 96-well plates. Results indicate a numerical increase in cell proliferation when exosomes from high-risk cows were applied compared with those from low-risk cows. Consistent with an effect on cell proliferation, quantitative reverse transcriptase PCR indicated a trend for upregulation of 3 proinflammatory genes (granulocyte colony-stimulating factor, ciliary neurotrophic factor, and CD27 ligand) with the application of high-risk exosomes, which are involved in cellular growth and survival. Proteomic analysis indicated 2 proteins in the low-risk group that were not identified in the high-risk group (endoplasmin and catalase), which may also be indicative of the metabolic state of origin. It is likely that the metabolic state of the transition cow affects cellular function through exosomal messaging; however, more in-depth research into cross-talk between exosomes and target cells is required to determine whether exosomes influence Madin-Darby bovine kidney cells in this manner.

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Preterm delivery is a major obstetric health problem contributing to poor neonatal outcome including low birth weight, respiratory distress syndrome, gastrointestinal, immunologic, central nervous system, hearing, and vision problems. Worldwide, approximately 15 million babies are born prematurely each year. The critical question which remains is how to identify women destined to deliver preterm from those who will achieve a term delivery. Prostaglandins, in all mammals, are important in the parturient process. Increased intrauterine prostaglandin production is associated with labor and in fact prostaglandin E2 (PGE2) or analogs are widely used clinically for cervical ripening and labor induction. Measurements of circulating eicosanoids have been problematic because of the rapid and major clearance by the lungs and then kidneys resulting in very low concentrations in plasma. Moreover, since eicosanoids are produced by all mammalian tissues, the sources of the measured eicosanoids are unknown. Our understanding of how cells communicate has undergone a paradigm shift with the recognition of the role of exosomes in intercellular signaling. Recent publications have identified enzymes and products of arachidonic acid metabolism (eicosanoids) within exosomes. This review will explore the potential roles of exosomes in eicosanoid functions that are critical in preterm labor and delivery.

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An increase in intrauterine prostaglandin production is critical for the onset and progression of labor in women and indeed all mammalian species studied. Endocannabinoids can act as substrates for enzymes of the prostaglandin biosynthetic pathways and can be utilized to generate other related compounds such as prostamides. The end products are indistinguishable by radioimmunoassay. We have separated such compounds by mass spectrometry. We now show that inflammatory stimuli such as LPS and proinflammatory cytokines act differentially on these pathways in human choriodecidua and preferentially create drive through to prostaglandin end products. These findings create doubt about the interpretation of data on prostaglandin biosynthesis in intrauterine tissues from pregnant women especially in the presence of an infection. The possibility is raised that separation of these products might reduce variability in results and lead to potential uses for their measurement in the diagnosis of preterm labor.

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INTRODUCTION: Gestational diabetes mellitus (GDM) is characterised by maternal glucose intolerance and insulin resistance during pregnancy. Myostatin, initially identified as a negative regulator of muscle development may also function in the regulation of placental development and glucose uptake. Myostatin expression in placentae of GDM complicated pregnancies is unknown. However, higher myostatin levels occur in placentae of pregnancies complicated with preeclampsia. We hypothesise that myostatin will be differentially expressed in GDM complicated pregnancies. METHODS: Myostatin concentrations (ELISA) were evaluated in plasma of presymptomatic women who later developed GDM and compared to plasma of normal glucose tolerant (NGT) women. Furthermore, myostatin protein expression (Western blot) was studied in placentae of pregnant women with GDM (treated with diet or insulin) compared to placentae of NGT women. RESULTS: No significant difference in myostatin concentration was seen in plasma of pre-symptomatic GDM women compared to NGT women. In placenta significant differences in myostatin protein expressions (higher precursor; p < 0.05and lower dimer: p < 0.005) were observed in GDM complicated compared to NGT pregnancies. Furthermore, placentae of GDM women treated with insulin compared to diet have higher dimer (p < 0.005) and lower precursor (p < 0.05). Compared to lean women, placentae of obese NGT women were lower in myostatin dimer expression (p < 0.05). DISCUSSION: Myostatin expression in placental tissue is altered under stress conditions (e.g. obesity and abnormal glucose metabolism) found in pregnancies complicated with GDM. We hypothesise that myostatin is active in these placentae and could affect glucose homoeostasis and/or cytokine production thereby altering the function of the placenta.

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INTRODUCTION: Labour and delivery are processes associated with inflammation within intrauterine and cervical tissues. The mechanisms that induce labour-associated changes and, in particular, the role of microRNAs (miRNAs) remain to be elucidated. MiRNAs are small non-coding RNAs that repress gene expression via mRNA degradation and translational repression. Let-7 miRNAs are negatively regulated by RNA-binding protein, Lin28, and both function downstream of NF-κB signalling. In non-gestational tissues, let-7 and Lin28 reportedy function as negative and positive regulators of IL-6 expression. We hypothesised that labour-associated inflammation involves the downregulation of let-7 miRNAs and upregulation of Lin28 expression. AIM: To determine the expression of Lin28 protein and let-7 miRNA in human gestational tissue obtained before and after labour. METHOD: Gestational tissues were collected from women at term by Caesarean section with and without labour and following normal vaginal delivery (n = 6 per group). Protein and RNA was extracted and Lin28 and let-7 miRNA expression was measured by Western blotting and real-time PCR. RESULTS: The data obtained established that let-7 miRNA and Lin28 display tissue-specific expression: Lin28 was strongly expressed in the placenta and choriodecidua, but not measurable in amnion; and let-7b and -7c expression were significantly lower in choriodecidua compare to placenta and amnion, whereas the amnion expressed less let-7d and -7f than other tissues. CONCLUSION: While the expression of Lin28 protein and let-7 miRNA did not vary significantly with labour onset and delivery, changes in their bioactivity and impact on nuclear signalling pathways in human gestational tissues remain to be established.

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When initially evaluating picture archiving communication systems (PACS) many radiologists try to duplicate the film environment and believe that multiple monitors are required to maintain the productivity of the radiologist. The authors were under the same impression initially but found that they underwent a paradigm shift over a period of time. This report documents the evolution that the radiologists underwent. The author's department consists of 28 diagnostic radiologists and 21 residents who actively read cases on a PACS. The department has been filmless for 6 months, although they have been reading soft copy films for 2 years. All modalities except mammography are included. The authors conducted interviews with both attending radiologists and residents to evaluate the change in methodology from the preconceptions to initial use to current use. The number and kind of monitors preferred for plain films, ultrasound scan, computed tomography (CT), and magnetic resonance imaging (MRI) were recorded. Additionally, viewing methods of different modalities were discussed. The authors found that there was a decrease in the number of monitors from preconceptions to actual use. Furthermore, to a lesser degree, there is a reduction of monitors used initially to that which is currently being used. The style of viewing cross-sectional images has changed. There has been a decrease in the number of images displayed on each monitor. The use of the roller ball on the mouse has affected this viewing style. Changing from a film-based reading environment to PACS environment not only brings about change in the overall technology in image delivery but also in the viewing techniques by radiologists. At our institution we have evolved from initially expecting to use 4 monitors all the time to actually preferring 2 monitors and occasionally 1 monitor to view images. Presentation software and viewing aids such as the roller ball on a mouse for viewing CTs in stack mode are key contributions to this paradigm shift. The decrease in monitors makes PACS more affordable and will allow further penetration of filmless radiology. The authors have found that after using PACS, radiologists prefer using 2 monitors. The style of reading films has changed with experience. Hospitals that plan to purchase PACS should consider this and ensure that the vendor has presentation software that optimizes the 2 monitor system.

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OBJECTIVE: The purpose of our study was to determine the degree of interobserver variability and correlation between MR cholangiopancreatography and endoscopic retrograde cholangiopancreatography (ERCP) for the presence of bile duct strictures in patients with primary sclerosing cholangitis. MATERIALS AND METHODS: For this retrospective study involving 26 patients with primary sclerosing cholangitis, 31 MR cholangiopancreatograms were compared with 30 endoscopic retrograde cholangiopancreatograms. The MR cholangiopancreatograms were independently interpreted by two abdominal radiologists in a blinded, randomized manner for overall image quality, extent of ductal visualization, and the presence and location of bile duct strictures. Unweighted multirater kappa coefficient values were estimated for each comparison. RESULTS: Visualization of more than 50% of the expected ductal length was possible in the extrahepatic, central intrahepatic, and peripheral intrahepatic bile ducts in 99%, 88%, and 69% of the MR cholangiopancreatograms and 100%, 86%, and 52% of the endoscopic retrograde cholangiopancreatograms, respectively. Strictures were detected in the extrahepatic, central, and peripheral ducts in 53%, 68%, and 87% of the MR cholangiopancreatograms and 73%, 67%, and 63% of the endoscopic retrograde cholangiopancreatograms, respectively. The interobserver agreement for stricture detection was 61% for MR cholangiopancreatography and 76% for ERCP. MR cholangiopancreatographic findings were consistent with ERCP findings for the presence of strictures in 69% of the cases. CONCLUSION: In patients with primary sclerosing cholangitis, MR cholangiopancreatography better shows the bile ducts and can depict more strictures, especially of the peripheral intrahepatic ducts, than ERCP. MR cholangiopancreatography can be used to noninvasively diagnose and follow up patients with primary sclerosing cholangitis.

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PURPOSE: In this manuscript, we present our initial experience with MRI of the abdomen at 8 T of canine subjects both alive and dead. Our hypothesis is that abdominal imaging at 8 T should be possible and should demonstrate unique information. To our knowledge, this is the first description of imaging characteristics of the abdomen at such field strengths using a human MR scanner. METHOD: An 8 T, 80 cm magnet housed in our department since 1998 was used for our study. GRE and rapid acquisition by relaxation enhancement (RARE) pulse sequences were selected to give reasonable slice profiles with relatively low power. Three dogs were imaged alive and after being killed. RESULTS: Our initial results show excellent signal-to-noise ratio and good RF penetration. Structures in the center of the abdomen were well visualized. Homogeneous signal was noted throughout each image without dielectric resonance artifact. Magnetic susceptibility artifacts were most severe on the GRE sequences. On the GRE sequences, the images appeared relatively T2 weighted. Signal voids were seen due to gas in the lung and bowel and susceptibility artifact at subcutaneous fat-muscle boundaries. The liver and spleen showed similar signal intensity, hypointense to subcutaneous muscle at low TE values. There was little internal anatomy of the liver or spleen visible except for the vessels. The kidney, in contrast, demonstrated very good internal structure with visualization of the cortex and medulla. Linear signal voids were depicted in the expected location of normal renal vascular anatomy on the GRE sequences. On the RARE sequences, the images also appeared T2 weighted. Magnetic susceptibility artifacts at subcutaneous fat-muscle boundaries were absent. Signal voids were noted in vessels with blood flow and gas. The liver and spleen were of similar signal intensity and slightly hypointense to muscle. The kidney and pancreas were of higher signal intensity than liver and subcutaneous muscle. The gallbladder wall demonstrated a striated pattern of two layers, with an inner hypointense and an outer hyperintense layer on the RARE sequence. The gastric wall demonstrated a striated pattern of five layers on the RARE sequence. CONCLUSION: Images of the dog abdomen with the world's first ultra high field 8 T magnet show robust image quality and excellent spatial resolution. Image contrast is greatest on the RARE sequence, and susceptibility artifact is strongest on the GRE sequence.

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Mangafodipir trisodium (Teslascan), a hepatobiliary contrast agent, has the potential of providing functional biliary imaging similar to hepatobiliary scintigraphy. To our knowledge. the potential role of this biliary contrast agent in the detection of bile duct leaks has not been reported. In this case report, we report the first case of a bile duct leak diagnosed with enhanced MRI with mangafodipir trisodium in a patient following laparoscopic cholecystectomy. Our case illustrates that functional MR cholangiography images can be successfully acquired by using a post-mangafodipir fat-suppressed GRE technique and that bile duct leaks can be detected.

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The appearance of the cirrhotic liver on computed tomography can be difficult to evaluate and can frustrate the radiologist distinguishing benign from malignant lesions. Hepatic edema, fibrosis, atrophy, and vascular abnormalities are common in the cirrhotic liver and produce derangements in morphology, attenuation, and perfusion, limiting the accurate characterization of hepatic masses. With the development of fast magnetic resonance (MR) sequences and dynamic postgadolinium-enhanced imaging, most hepatic lesions with uncertain etiology on computed tomography can be accurately characterized on MR imaging. We describe MR imaging techniques useful for imaging cirrhosis and its complications. We also illustrate the spectrum of findings in the cirrhotic liver on dynamic gadolinium-enhanced MR imaging, including reticular and confluent fibrosis, fatty infiltration, hemochromatosis, regenerating nodules, dysplastic nodules, hepatocellular carcinoma, and sequela of portal hypertension.

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