Joint total variation-based reconstruction of multiparametric magnetic resonance images for mapping tissue types.

Multispectral analysis of coregistered multiparametric magnetic resonance (MR) images provides a powerful method for tissue phenotyping and segmentation. Acquisition of a sufficiently varied set of multicontrast MR images and parameter maps to objectively define multiple normal and pathologic tissue types can require long scan times. Accelerated MRI on clinical scanners with multichannel receivers exploits techniques such as parallel imaging, while accelerated preclinical MRI scanning must rely on alternate approaches. In this work, tumor-bearing mice were imaged at 7 T to acquire k-space data corresponding to a series of images with varying T1-, T2- and T2*-weighting. A joint reconstruction framework is proposed to reconstruct a series of T1-weighted images and corresponding T1 maps simultaneously from undersampled Cartesian k-space data. The ambiguity introduced by undersampling was resolved by using model-based constraints and structural information from a reference fully sampled image as the joint total variation prior. This process was repeated to reconstruct T2-weighted and T2*-weighted images and corresponding maps of T2 and T2* from undersampled Cartesian k-space data. Validation of the reconstructed images and parameter maps was carried out by computing tissue-type maps, as well as maps of the proton density fat fraction (PDFF), proton density water fraction (PDwF), fat relaxation rate ( R2f*) and water relaxation rate ( R2w*) from the reconstructed data, and comparing them with ground truth (GT) equivalents. Tissue-type maps computed using 18% k-space data were visually similar to GT tissue-type maps, with dice coefficients ranging from 0.43 to 0.73 for tumor, fluid adipose and muscle tissue types. The mean T1 and T2 values within each tissue type computed using only 18% k-space data were within 8%-10% of the GT values from fully sampled data. The PDFF and PDwF maps computed using 27% k-space data were within 3%-15% of GT values and showed good agreement with the expected values for the four tissue types.

Effects of feeding OmniGen-AF® on superovulatory response in donor beef cows: I. Serum progesterone and cortisol, embryo recovery and quality.

Optimal results in cattle embryo transfer are limited by the variation in ova recovery, fertilization rate and embryo quality experienced with superovulation. Inflammation and immune dysregulation may be contributing factors. This study, evaluated feeding OmniGen-AF® (OG), a nutritional supplement that reduces inflammation and supports immune health, on superovulatory response and serum progesterone and cortisol concentrations in embryo donors treated with two different doses of Folltropin®-V (FSH). Angus cross-bred beef cows (n = 24) were assigned to four groups, fed OG at 0 or 56 g/animal/day for 49 days and were treated with 200 or 400 mg FSH to induce superovulation. Treatments for superovulation started after feeding OG for 28 days and ova were non-surgically recovered 7 days after estrus and graded for quality. More transferrable embryos (P < 0.05) were recovered from cows fed 56 g OG and treated with 400 compared with 200 mg FSH. Percent degenerate embryos recovered from cows treated with the 400 mg FSH dose was threefold greater (P < 0.05) when fed no OG compared with 56 g OG. Serum progesterone on day of embryo collection was greater (P < 0.05) in OG-supplemented cows and cows treated with 200 mg FSH. Serum cortisol was not affected (P > 0.10) by FSH dose or OG-feeding, but was greatest (P <  0.05) on Days 0 and 42 of the feeding period. In summary, the improvement in embryo quality with OG-feeding may relate to a greater serum progesterone concentration.

Endometriosis of the appendix presenting as acute appendicitis with unusual appearance.

INTRODUCTION: Endometriosis of the appendix is an uncommon mimicker of acute appendicitis which makes for a diagnostic dilemma. PRESENTATION OF CASE: We present a rare case of a menstruating woman presenting with classic symptoms of appendicitis, without the characteristic inflammatory changes seen on laparoscopy consistent with appendicitis. Instead, the appendix appeared unusually contracted on itself. Pathologic review of the appendix revealed microscopic findings of endometriosis. DISCUSSION: We theorize the growth and shedding of the endometrial tissue during menstruation caused compression of the neural plexi in the wall of the appendix leading to the presentation mimicking acute appendicitis. CONCLUSION: Given the potential for endometrial appendicitis, we propose appendectomy in reproductive age female patients with right lower quadrant pain, regardless of appendix appearance on laparoscopy.

Phosphorylation of a constrained azacyclic FTY720 analog enhances anti-leukemic activity without inducing S1P receptor activation.

The frequency of poor outcomes in relapsed leukemia patients underscores the need for novel therapeutic approaches. The Food and Drug Administration-approved immunosuppressant FTY720 limits leukemia progression by activating protein phosphatase 2A and restricting nutrient access. Unfortunately, FTY720 cannot be re-purposed for use in cancer patients due to on-target toxicity associated with S1P receptor activation at the elevated, anti-neoplastic dose. Here we show that the constrained azacyclic FTY720 analog SH-RF-177 lacks S1P receptor activity but maintains anti-leukemic activity in vitro and in vivo. SH-RF-177 was not only more potent than FTY720, but killed via a distinct mechanism. Phosphorylation is dispensable for FTY720's anti-leukemic actions. However, chemical biology and genetic approaches demonstrated that the sphingosine kinase 2 (SPHK2)-mediated phosphorylation of SH-RF-177 led to engagement of a pro-apoptotic target and increased potency. The cytotoxicity of membrane-permeant FTY720 phosphonate esters suggests that the enhanced potency of SH-RF-177 stems from its more efficient phosphorylation. The tight inverse correlation between SH-RF-177 IC50 and SPHK2 mRNA expression suggests a useful biomarker for SH-RF-177 sensitivity. In summary, these studies indicate that FTY720 analogs that are efficiently phosphorylated but fail to activate S1P receptors may be superior anti-leukemic agents compared to compounds that avoid cardiotoxicity by eliminating phosphorylation.

Alkaline ceramidase 3 deficiency aggravates colitis and colitis-associated tumorigenesis in mice by hyperactivating the innate immune system.

Increasing studies suggest that ceramides differing in acyl chain length and/or degree of unsaturation have distinct roles in mediating biological responses. However, still much remains unclear about regulation and role of distinct ceramide species in the immune response. Here, we demonstrate that alkaline ceramidase 3 (Acer3) mediates the immune response by regulating the levels of C18:1-ceramide in cells of the innate immune system and that Acer3 deficiency aggravates colitis in a murine model by augmenting the expression of pro-inflammatory cytokines in myeloid and colonic epithelial cells (CECs). According to the NCBI Gene Expression Omnibus (GEO) database, ACER3 is downregulated in immune cells in response to lipopolysaccharides (LPS), a potent inducer of the innate immune response. Consistent with these data, we demonstrated that LPS downregulated both Acer3 mRNA levels and its enzymatic activity while elevating C(18:1)-ceramide, a substrate of Acer3, in murine immune cells or CECs. Knocking out Acer3 enhanced the elevation of C(18:1)-ceramide and the expression of pro-inflammatory cytokines in immune cells and CECs in response to LPS challenge. Similar to Acer3 knockout, treatment with C(18:1)-ceramide, but not C18:0-ceramide, potentiated LPS-induced expression of pro-inflammatory cytokines in immune cells. In the mouse model of dextran sulfate sodium-induced colitis, Acer3 deficiency augmented colitis-associated elevation of colonic C(18:1)-ceramide and pro-inflammatory cytokines. Acer3 deficiency aggravated diarrhea, rectal bleeding, weight loss and mortality. Pathological analyses revealed that Acer3 deficiency augmented colonic shortening, immune cell infiltration, colonic epithelial damage and systemic inflammation. Acer3 deficiency also aggravated colonic dysplasia in a mouse model of colitis-associated colorectal cancer. Taken together, these results suggest that Acer3 has an important anti-inflammatory role by suppressing cellular or tissue C(18:1)-ceramide, a potent pro-inflammatory bioactive lipid and that dysregulation of ACER3 and C(18:1)-ceramide may contribute to the pathogenesis of inflammatory diseases including cancer.

Global identification of genes targeted by DNMT3b for epigenetic silencing in lung cancer.

The maintenance cytosine DNA methyltransferase DNMT1 and de novo methyltransferase DNMT3b cooperate to establish aberrant DNA methylation and chromatin complexes to repress gene transcription during cancer development. The expression of DNMT3b was constitutively increased 5-20-fold in hTERT/CDK4-immortalized human bronchial epithelial cells (HBECs) before treatment with low doses of tobacco carcinogens. Overexpression of DNMT3b increased and accelerated carcinogen-induced transformation. Genome-wide profiling of transformed HBECs identified 143 DNMT3b-target genes, many of which were transcriptionally regulated by the polycomb repressive complex 2 (PRC2) complex and silenced through aberrant methylation in non-small-cell lung cancer cell lines. Two genes studied in detail, MAL and OLIG2, were silenced during transformation, initially through enrichment for H3K27me3 and H3K9me2, commonly methylated in lung cancer, and exert tumor suppressor effects in vivo through modulating cancer-related pathways. Re-expression of MAL and OLIG2 to physiological levels dramatically reduced the growth of lung tumor xenografts. Our results identify a key role for DNMT3b in the earliest stages of initiation and provide a comprehensive catalog of genes targeted for silencing by this methyltransferase in non-small-cell lung cancer.

The negative predictive value of spiral computed tomography for the diagnosis of pulmonary embolism in patients with nondiagnostic ventilation-perfusion scans.

BACKGROUND: There is no noninvasive method to rule out pulmonary embolism when the clinical suspicion for pulmonary embolism is high. We did a prospective observational study to determine the negative predictive value of spiral computed tomography (CT) in this situation. METHODS: We performed spiral CT scans of the thorax in consecutive patients with high clinical suspicion of pulmonary embolism with intermediate or low probability ventilation-perfusion scans. Patients with negative or indeterminate spiral CT results had conventional angiography at the discretion of the attending physician. Only patients with positive spiral CT results or positive conventional angiograms were treated. All patients were observed for 6 months for evidence of venous thromboembolic disease. Clinical outcome without treatment or the results of conventional angiography were used as reference standards. False-negative results were defined as a negative spiral CT with a positive conventional angiogram or any diagnosis of venous thromboembolism within 6 months. RESULTS: Among the 103 patients who were studied, spiral CT scans were positive in 22 patients, indeterminate in 10 patients, and negative in 71 patients. Twenty-seven (26%) patients had pulmonary embolism by clinical outcome, including 3 of the 71 patients with negative spiral CT scans and 2 of the 10 patients with indeterminate scans. A negative spiral CT result had a likelihood ratio of 0.12 (95% confidence interval [CI]: 0.04 to 0.35) with a negative predictive value of 96% (95% CI: 88% to 99%). Using conventional angiography only as the reference standard, a negative spiral CT result had a likelihood ratio of 0.08 (95% CI: 0.02 to 0.31) and a negative predictive value of 93% (95% CI: 77% to 98%). CONCLUSIONS: Spiral CT has a high negative predictive value for pulmonary embolism and may replace conventional angiography in the workup of pulmonary embolism. Patients with indeterminate spiral CT results should be considered for conventional angiography.

Diastolic function in neonates after the arterial switch operation: effects of positive pressure ventilation and inspiratory time.

OBJECTIVE: To determine the effects positive pressure ventilation have on left ventricular diastolic function in neonates after the arterial switch operation. DESIGN: Prospective case series. SETTING: Pediatric cardiac and multidisciplinary intensive care units in two university-affiliated children's hospitals. PATIENTS AND PARTICIPANTS: The patient population consisted of 12 neonates weighing 2.5-4.2 kg with D-transposition of the great arteries (DTGA) who underwent arterial switch operation. INTERVENTIONS: All patients were mechanically ventilated in a volume-targeted mode with a square wave flow pattern. The positive end-expiratory pressure was held constant. A long inspiratory time was set by extending it over three cardiac cycles. MEASUREMENTS AND RESULTS: Pulsed Doppler measurements of left ventricular diastolic function were performed during the following cardiac cycles: (1) the last diastolic period of expiration (E(L)), (2) first, second and third diastolic periods of inspiration (I1, I2, I3) and (3) the first diastolic period of expiration (E(F)). Doppler measurements of peak E wave, peak A wave, E area/A area, E area fraction, A area fraction, 0.33 area fraction and the deceleration time were made. Doppler tracings were digitized and the data obtained from three sequential study periods were averaged. Data were statistically analyzed using the repeated measures analysis of variance procedure. During (I1), there was a 21% increase in the peak E wave (0.53+/-0.06 vs 0.64+/-0.08 m/s, p < 0.01) and 28% increase in peak A wave (0.47+/-0.07 vs 0.60+/-0.08 m/s, p < 0.01) compared to (E(L)). There was a 24% increase in total area under the E and A waves when I1 was compared to E(L) (0.059+/-0.008 vs 0.073+/-0.009, p < 0.01) and there was no change in mitral valve deceleration time. Compared to the initial diastolic period during inspiration (I1), the third diastolic period during inspiration (I3) had a 38% decrease in peak E (0.64+/-0.08 vs 0.40+/-0.05 m/s, p < 0.01) and 33% decrease in peak A (0.60+/-0.09 vs 0.40+/-0.05 m/s, p < 0.01). In addition, there was a 16% reduction in total area under the E and A waves (0.073+/-0.009 vs 0.061+/-0.008, p < 0.01). There were no changes in the other diastolic indexes that reflect changes in ventricular compliance or relaxation. CONCLUSIONS: In neonates with transposition of the great arteries (TGA) after the arterial switch operation, positive pressure ventilation augments left ventricular filling during the early phase of inspiration. Prolonging the inspiratory time over three cardiac cycles results in a reduction in left ventricular filling during the third diastolic period. There were no changes in the other diastolic indexes that reflect changes in ventricular compliance or relaxation.

Effects of kappa opioids in the inflamed rat colon.

The objective of this study was to examine the antinociceptive effects of peripherally restricted kappa-opioid receptor agonists (ORAs) in a rat model of inflammatory bowel disease produced by intracolonic instillation of trinitrobenzine sulfonic acid (TNBS). Antinociceptive effects of mu-(morphine) and kappa-ORAs (EMD 61,753 and ICI 204,488) were evaluated in a behavioral model of visceral nociception. The effects of these agonists and a delta-ORA (SNC 80) on responses of pelvic nerve afferent fibers innervating the colon were also tested. In the behavioral study, systemic injections of morphine and both kappa-ORAs dose-dependently inhibited the visceromotor response to colorectal distension in rats with uninflamed or inflamed colons. The inhibitory effects of kappa-ORAs, but not morphine, were significantly greater in rats with colons inflamed 4 days previously by TNBS. A mu-receptor-selective dose (30 microg/kg) of naloxone methiodide (NLXM) blocked the inhibitory effect of morphine, but not of EMD 61,753. In the single-fiber study, neither morphine nor the delta-ORA SNC 80 attenuated the responses of pelvic nerve afferent fibers, whereas kappa-ORAs dose-dependently inhibited responses of pelvic nerve afferent fibers with significantly greater potency in the inflamed colon. Pretreatment with a non-opioid receptor-selective dose (2 mg/kg) of NLXM produced a rightward shift in the dose-response function of EMD 61,753. The greater potency of kappa-ORAs in the TNBS-inflamed condition suggests a peripheral upregulation of kappa-opioid receptors in colonic inflammation.

In vitro analysis of regurgitant fraction using Doppler power-weighted sum of velocities.

The power-weighted sum of velocities (PWS) is the sum of each velocity component of the Doppler signal multiplied by its power. The purpose of this study was to determine (1) whether PWS is linearly related to volume flow and (2) whether PWS can predict the regurgitant fraction in an in vitro pulsatile flow system simulating aortic regurgitation. Doppler analysis of aortic flow was performed with an intact valve and two regurgitant valves. For each valve a linear relation between the forward flow PWS and forward flow volume was demonstrated, with excellent correlation (r = 0.99). For the valves with regurgitant orifices, the values for the PWS-derived regurgitant fraction were compared with measured regurgitant fraction. A fair correlation was demonstrated (r = 0.59), with low accuracy in prediction (error 44% +/- 24%). The PWS was inaccurate in predicting flow ratios in our in vitro system despite the strong relation with forward flow volume. The error incurred may be due to effects of filters that remove low velocity and low amplitude information.

Reduced chlorhexidine tooth stain coverage by sequential administration of monoperoxyphthalic acid in the beagle dog.

Extrinsic tooth stain is sometimes a side-effect observed with the chronic use of chlorhexidine-containing oral care products. This chlorhexidine (CHX)-induced tooth staining is observed both in humans as well as in experimental animals. The present study tested the hypothesis that the sequential administration of monoperoxyphthalic acid (MPA) will reduce the development of chlorhexidine-induced tooth stain coverage in a beagle dog. For this study, dogs were treated with 30 mL of mouthrinse b.i.d. for 28 days. The following treatment groups ( = 12 dogs/group) were tested: water (negative control), 1.0% MPA rinse, 0.12% CHX rinse, and 1.0% MPA rinse followed by 0.12% CHX rinse. The sequential dosing of 1.0% MPA followed by 0.12+ CHX resulted in significantly (p < or = 0.05) less tooth stain than when dogs were dosed with 0.12% CHX alone. Additionally, the sequential dosing of MPA followed by chlorhexidine resulted in a 75% reduction in plaque formation in this model, which was significantly different (p < or = 0.05) from results with either treatment alone. A further study demonstrated that similar results could be obtained when MPA plus CHX treatment alone. A further study demonstrated that similar results could be obtained when MPA plus CHX treatments were separated by approximately a five-hour period in an AM and PM schedule. We conclude that the oxidizer, MPA, can significantly reduce tooth stain induced by CHX while enhancing its antiplaque and antigingivitis activity.

Doppler systemic venous flow patterns: changes in children with mild/moderate pulmonic stenosis.

To determine the usefulness of systemic venous flow patterns in patients with mild/moderate right ventricular hypertension, 17 patients with isolated mild/moderate pulmonic stenosis and 17 age-matched normal children were evaluated with pulsed Doppler echocardiography. Tricuspid valve, superior vena caval, and hepatic vein pulsed Doppler recordings were obtained with simultaneous respirometry and electrocardiography. Peak velocities and velocity-time integrals were measured for Doppler signals corresponding with ventricular systole, ventricular diastole, ventricular end systole, and atrial systole. The groups were similar in weight, heart rate, tricuspid inflow Doppler echocardiograms, and cardiac indexes. Compared with normal subjects, patients showed changes in respiratory variation for some superior vena caval and hepatic vein indexes. In addition, hepatic vein measurements made at ventricular end systole were significantly lower and measurements made at atrial systole were significantly higher in patients than in normal subjects. These changes in systemic venous flow patterns may provide a sensitive indicator of early right-sided heart dysfunction.

Human immunodeficiency virus type 1 TAR element revertant viruses define RNA structures required for efficient viral gene expression and replication.

The TAR element is a viral regulatory element extending from +1 to +60 in the human immunodeficiency virus type 1 (HIV-1) long terminal repeat, which is critical for activation by the transactivator protein Tat. Jurkat cell lines chronically infected with viruses containing HIV-1 TAR element mutations are extremely defective for both gene expression and replication. We previously demonstrated that viruses containing mutations of the TAR RNA stem, bulge, or loop structures have 200- to 5,000-fold-reduced levels of gene expression compared with lymphoid cells harboring wild-type virus. In this study, we characterized several Jurkat cell lines infected with TAR element mutant viruses which spontaneously produced culture supernatants with wild-type-like levels of reverse transcriptase activity. These viral supernatants were used to infect Jurkat cells, and following PCR amplification of the viral long terminal repeats, their DNA sequences were analyzed. This analysis demonstrated that revertant viruses isolated from these cell lines retained the original TAR mutations but also contained additional compensatory mutations within TAR. In gel retardation analysis, recombinant Tat protein bound to higher levels to in vitro-transcribed revertant TAR RNAs than the original TAR RNA mutants. Both the original and revertant TAR elements were inserted into both chloramphenicol acetyltransferase reporter and HIV-1 proviral constructs and assayed following transfection of Jurkat cells. Constructs containing revertant TAR element mutations were capable of strong activation by Tat in contrast to constructs containing the original TAR mutations. Analysis of the secondary structure of TAR RNA sequences suggested that TAR RNA structures which differed from that of wild-type TAR were still capable of strong activation in response to Tat. These results further define critical sequences in TAR RNA that are required for tat activation. In addition, since TAR structures with lower free energy that preserve the loop and bulge structures may be favored over fully formed TAR RNA with higher stable free energy, these results implicate nascent RNA rather than the fully formed TAR RNA structure as the target for tat activation.

In-vivo analysis of hpv e7 protein association with prb, p107 and p130.

The two-hybrid system was used to detect interactions in vivo between HPV E7 and three 'Rb-like proteins', pRb, p107 and p130. The association between pE7 and pRb parallel the oncogenic potential of the specific HPV types. In contrast, the interaction between pE7 and p107 or p130 differ. While the HPV 16 E7 protein associates with the 'Rb-like' proteins strongly, both HPV 18 and 6b E7 proteins bind more weakly. We tested several HPV 6 E7 mutants carrying single amino acid mutations. Substitution of the glycine at position 22 with an aspartate was the only mutation capable of increasing the ability of HPV 6 E7 protein to bind pRb. However, association with p107 and p130 by the HPV 6 E7 protein was also increased by mutation of the arginine at position 4 with an aspartate. These data suggest that pRb, p107 and p130 interact with similar but non-identical domains of pE7. In addition, we used amphotropic retroviruses encoding the HPV 18 E6 and the different E7 genes to analyze their immortalizing activity. The wild-type HPV Is and 16 E7 genes complemented the HPV 18 E6 gene to immortalize human keratinocytes. In comparison, none of the cells infected with HPV 6 E7, wildtype or mutant- encoding retroviruses, became immortal. Thus, our data suggest that HPV 6 E7 lacks a property independent of pRb-association which is required for immortalization of human keratinocytes.

An exact statistical method for comparing topographic maps, with any number of subjects and electrodes.

Statistical methods for testing differences between neural images (e.g., PET, MRI or EEG maps) are problematic because they require (1) an untenable assumption of data sphericity and (2) a high subject to electrode ratio. We propose and demonstrate an exact and distribution-free method of significance testing which avoids the sphericity assumption and may be computed for any combination of electrode and subject numbers. While this procedure is rigorously rooted in permutation test theory, it is intuitively comprehensible. The sensitivity of the permutation test to graded changes in dipole location for systematically varying levels of signal/noise ratio, intersubject variability and number of subjects was demonstrated through a simulation of 70 different conditions, generating 5,000 different data sets for each condition. Data sets were simulated from a homogeneous single-shell dipole model. For noise levels commonly encountered in evoked potential studies and for situations where the number of subjects was less than the number of electrodes, the permutation test was very sensitive to a change in dipole location of less than 0.75 cm. This method is especially sensitive to localized changes that would be "washed-out" by more traditional methods of analysis. It is superior to all previous methods of statistical analysis for comparing topographical maps, because the test is exact, there is no assumption of a multivariate normal distribution or of the correlation structure of the data requiring correction, the test can be tailored to the specific experimental hypotheses rather than allowing the statistical tests to limit the experimental design, and there is no limitation on the number of electrodes that can be simultaneously analyzed.

Echocardiographic guidance during placement of the buttoned double-disk device for atrial septal defect closure.

The usefulness of two-dimensional and Doppler echocardiography during buttoned double-disk device closure of an atrial septal defect was evaluated in 20 consecutive patients at the time of interventional catheterization. Transesophageal echocardiography was used in 11 patients (ages 5 to 62 years, weights 20 to 91 kg). Because of the size of the available transesophageal echo probe, transthoracic echocardiography was used in the remaining 9 patients (ages 4 to 5.5 years, weights 14 to 21 kg). In the transesophageal echo group, 1 patient was found to have no atrial septal defect despite a previous diagnosis by transthoracic echocardiography, 3 patients had atrial septal defects too large for closure despite attempts in 2, and 7 patients had transesophageal echo guided device placement. All of these 7 patients had small residual shunts by color Doppler, 2 had unusual arm positions, and 2 had surgical removal of the device due to embolization to the pulmonary artery in 1 and failure to obtain close approximation of the occluder and counteroccluder in 1. In the transthoracic echo group, 2 patients had atrial septal defects too large for closure, 1 patient had no femoral venous access, and 6 patients had transthoracic echo guided device placement. All of these 6 patients had small residual shunts by color Doppler and 3 of the 6 had unusual arm positions. For atrial septal defect sizing, transesophageal echo measurements correlated with catheter balloon size more closely than did transthoracic echo measurements (r 2 = 0.97 vs 0.86).(ABSTRACT TRUNCATED AT 250 WORDS)