High-resolution µ CT imaging for characterizing microcalcification detection performance in breast CT.

Purpose: To demonstrate the utility of high-resolution micro-computed tomography ( µ CT ) for determining ground-truth size and shape properties of calcium grains for evaluation of detection performance in breast CT (bCT). Approach: Calcium carbonate grains ( ∼ 200 µ m ) were suspended in 1% agar solution to emulate microcalcifications ( µ Calcs ) within a fibroglandular tissue background. Ground-truth imaging was performed on a commercial µ CT scanner and was used for assessing calcium-grain size and shape, and for generating µ Calc signal profiles. Calcium grains were placed within a realistic breast-shaped phantom and imaged on a prototype bCT system at 3- and 6-mGy mean glandular dose (MGD) levels, and the non-prewhitening detectability was assessed. Additionally, the µ CT -derived signal profiles were used in conjunction with the bCT system characterization (MTF and NPS) to obtain predictions of bCT detectability. Results: Estimated detectability of the calcium grains on the bCT system ranged from 2.5 to 10.6 for 3 mGy and from 3.8 to 15.3 for 6 mGy with large fractions of the grains meeting the Rose criterion for visibility. Segmentation of µ CT images based on morphological operations produced accurate results in terms of segmentation boundaries and segmented region size. A regression model linking bCT detectability to µ Calc parameters indicated significant effects of µ Calc size and vertical position within the breast phantom. Detectability using µ CT -derived detection templates and bCT statistical properties (MTF and NPS) were in good correspondence with those measured directly from bCT ( R 2 > 0.88 ). Conclusions: Parameters derived from µ CT ground-truth data were shown to produce useful characterizations of detectability when compared to estimates derived directly from bCT. Signal profiles derived from µ CT imaging can be used in conjunction with measured or hypothesized statistical properties to evaluate the performance of a system, or system component, that may not currently be available.

Modular Synthesis of Peptide-Based Single- and Multimodal Targeted Molecular Imaging Agents.

A practical, modular synthesis of targeted molecular imaging agents (TMIAs) containing near-infrared dyes for optical molecular imaging (OMI) or chelated metals for magnetic resonance imaging (MRI) and single-photon emission correlation tomography (SPECT) or positron emission tomography (PET) has been developed. In the method, imaging modules are formed early in the synthesis by attaching imaging agents to the side chain of protected lysines. These modules may be assembled to provide a given set of single- or dual-modal imaging agents, which may be conjugated in the last steps of the synthesis under mild conditions to linkers and targeting groups. A key discovery was the ability of a metal such as gadolinium, useful in MRI, to serve as a protecting group for the chelator, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA). It was further discovered that two lanthanide metals, La and Ce, can double as protecting groups and placeholder metals, which may be transmetalated under mild conditions by metals used for PET in the final step. The modular method enabled the synthesis of discrete targeted probes with two of the same or different dyes, two same or different metals, or mixtures of dyes and metals. The approach was exemplified by the synthesis of single- or dual-modal imaging modules for MRI-OMI, PET-OMI, and PET-MRI, followed by conjugation to the integrin-seeking peptide, c(RGDyK). For Gd modules, their efficacy for MRI was verified by measuring the NMR spin-lattice relaxivity. To validate functional imaging of TMIAs, dual-modal agents containing Cy5.5 were shown to target A549 cancer cells by confocal fluorescence microscopy.

A prototype Multi-X-ray-source array (MXA) for digital breast tomosynthesis.

The design and testing of a prototype Multi-X-ray-source Array (MXA) for digital breast tomosynthesis is reported. The MXA is comprised of an array of tungsten filament cathodes with focus cup grid-controlled modulation and a common rotating anode housed in a single vacuum envelope. Prototypes consisting of arrays of three-source elements and eleven-source-elements were fabricated and evaluated. The prototype sources demonstrated focal spot sizes of 0.3 mm at 45 kV with 50 mA. Measured x-ray spectra were consistent with the molybdenum anode employed, and the tube output (air kerma) was between 0.6 mGy/100 mAs at 20 kV and 17 mGy/100 mAs at 45 kV with a distance of 100 cm. HVL measurements ranged from 0.5 mm Al at 30 kV to 0.8 mm Al at 45 kV, and x-ray pulse widths were varied from 20 ms to 110 ms at operating frequencies ultimately to be limited by source turn-on/off times of ∼1 ms. Initial results of reconstructed tomographic data are presented.

High resolution microcalcification signal profiles for dedicated breast CT.

This study introduces a methodology for generating high resolution signal profiles of microcalcification (MC) grains for validating breast CT (bCT) systems. A physical MC phantom was constructed by suspending calcium carbonate grains in an agar solution emulating MCs in a fibroglandular tissue background. Additionally, small Teflon spheres (2.4 mm diameter) were embedded in the agar solution for the purpose of fiducial marking and assessment of segmentation accuracy. The MC phantom was imaged on a high resolution (34 µm) commercial small-bore µCT scanner at high dose, and the images were used as the gold-standard for assessing MC size and for generating high resolution signal profiles of each MC. High-dose bCT scans of the MC phantom suspended in-air were acquired using 1 × 1 binning mode (75 µm dexel pitch) by averaging three repeat scans to produce a single low-noise reconstruction of the MC phantom. The high resolution µCT volume data set was then registered with the corresponding bCT data set after correcting for the bCT system spatial resolution. Microcalcification signal profiles constructed using low-noise bCT images were found to be in good agreement with those generated using the µCT scanner with all differences < 10% within the VOI surrounding each MC. The MC signal profiles were used as detection templates for a non-prewhitening-matched-filter model observer for scans acquired in a realistic breast phantom at 3, 6, and 9 mGy mean glandular dose. MC detectability using signal templates derived from bCT were shown to be in good agreement with those generated using µCT.

Updated breast CT dose coefficients (DgNCT ) using patient-derived breast shapes and heterogeneous fibroglandular distributions.

PURPOSE: The purpose of this work was to generate uncompressed heterogeneous breast phantom models using size-dependent fibroglandular distributions derived from a large cohort of breast CT (bCT) datasets, and to compare differences in normalized glandular dose coefficients for bCT "DgNCT " when the realistic heterogeneous model is considered relative to the simple, homogeneous model used in the past. METHODS: A cohort of 274 segmented bCT datasets were used to quantify the fibroglandular tissue distribution within the breast parenchyma. Each dataset was interpolated to an isotropic voxel size of 0.25 mm and the breast center-of-mass was aligned for all coronal slices. Each aligned dataset was converted into two binarized volumes representing voxels containing only glandular tissue "G(x,y,z)" and voxels containing glandular or adipose tissue "AG(x,y,z)". The datasets were classified by volume in accordance with previously reported size-dependent, breast-shaped phantoms. Within the five groups - each containing on average 55 datasets, all G(x,y,z) and AG(x,y,z) volumes were summed separately representing the cumulative distribution of glandular tissue or breast parenchyma (glandular + adipose), respectively. G(x,y,z) was divided by AG(x,y,z) on a voxel-by-voxel basis resulting in a glandular fraction "GF(x,y,z)" distribution for each phantom size. The GF(x,y,z) distributions were used to construct heterogeneous mathematical phantoms for the small, median, and large breast sizes with a 1.5 mm skin thickness - based on previously reported measurements from bCT, and a 5 mm skin thickness for comparison with outdated assumptions of skin thickness. A subset of 15 bCT datasets from the cohort (five for each breast size) were used to construct voxelized patient models for validation of the heterogeneous phantom models. Monte Carlo techniques were used to estimate monoenergetic DgN(E)C T values for photon energies from 9 to 70 keV (in 1 keV intervals) using the mathematical phantoms composed of either heterogeneous or homogeneous breast parenchyma. Polyenergetic (pDgNCT ) coefficients were determined by weighting the DgN(E)C T values by x-ray spectra tuned to the beam characteristic of breast CT. Dose coefficients were compared between the two breast compositions for each volume class, breast density, and skin thickness. RESULTS: For photon energies ≲45 keV, the homogeneous model overestimates DgN(E) values relative to the realistic heterogeneous model sorted into five volume classes. The 5 mm skin thickness underestimates DgN(E) values relative to the realistic 1.5 mm thickness for lower energies and the differences diminish up to 70 keV. Averaged across all phantom sizes the homogeneous model overestimates pDgNCT by 5.7% and 23.3% for the 60 kV W/Cu and 49 kV W/Al spectra, respectively. The heterogeneous model was also found to be in agreement with the voxelized bCT patient models with pDgNCT differences less than 2.3% and 5.2% for the 60 kV W/Cu and 49 kV W/Al spectra, respectively, across all phantom sizes. CONCLUSION: Anatomically accurate heterogeneous phantom models were developed using bCT image-derived fibroglandular tissue distributions. These new models improve the accuracy of bCT dosimetry, and in conjunction with previous models for mammography, may help in providing a more universally accepted breast dosimetry model.

GPR18 Controls Reconstitution of Mouse Small Intestine Intraepithelial Lymphocytes following Bone Marrow Transplantation.

Specific G protein coupled receptors (GPRs) regulate the proper positioning, function, and development of immune lineage subsets. Here, we demonstrate that GPR18 regulates the reconstitution of intraepithelial lymphocytes (IELs) of the small intestine following bone marrow transplantation. Through analysis of transcriptional microarray data, we find that GPR18 is highly expressed in IELs, lymphoid progenitors, and mature follicular B cells. To establish the physiological role of this largely uncharacterized GPR, we generated Gpr18-/- mice. Despite high levels of GPR18 expression in specific hematopoietic progenitors, Gpr18-/- mice have no defects in lymphopoiesis or myelopoiesis. Moreover, antibody responses following immunization with hapten-protein conjugates or infection with West Nile virus are normal in Gpr18-/- mice. Steady-state numbers of IELs are also normal in Gpr18-/- mice. However, competitive bone marrow reconstitution experiments demonstrate that GPR18 is cell-intrinsically required for the optimal restoration of small intestine TCRγδ+ and TCRαß+ CD8αα+ IELs. In contrast, GPR18 is dispensable for the reconstitution of large intestine IELs. Moreover, Gpr18-/- bone marrow reconstitutes small intestine IELs similarly to controls in athymic recipients. Gpr18-/- chimeras show no changes in susceptibility to intestinal insults such as Citrobacter rodentium infections or graft versus host disease. These data reveal highly specific requirements for GPR18 in the development and reconstitution of thymus-derived intestinal IEL subsets in the steady-state and after bone marrow transplantation.

ADAM17 limits the expression of CSF1R on murine hematopoietic progenitors.

All-lymphoid progenitors (ALPs) yield few myeloid cells in vivo, but readily generate such cells in vitro. The basis for this difference remains unknown. We hypothesized that ALPs limit responsiveness to in vivo concentrations of myeloid-promoting cytokines by reducing expression of the corresponding receptors, potentially through posttranscriptional mechanisms. Consistent with such a mechanism, ALPs express higher levels of CSF1R transcripts than their upstream precursors, yet show limited cell-surface protein expression of colony-stimulating factor 1 receptor (CSF1R). All-lymphoid progenitors and other hematopoietic progenitors deficient in A disintegrin and metalloproteinase domain 17 (ADAM17), display elevated cell surface CSF1R expression. ADAM17(-/-) ALPs, however, fail to yield myeloid cells upon transplantation into irradiated recipients. Moreover, ADAM17(-/-) ALPs yield fewer macrophages in vitro than control ALPs at high concentrations of macrophage colony stimulating factor. Mice with hematopoietic-specific deletion of ADAM17 have normal numbers of myeloid and lymphoid progenitors and mature cells in vivo. These data demonstrate that ADAM17 limits CSF1R protein expression on hematopoietic progenitors, but that compensatory mechanisms prevent elevated CSF1R levels from altering lymphoid progenitor potential.

Blood pressure abnormalities in children with sickle cell anemia.

BACKGROUND: Kidney disease is an important cause of morbidity and mortality in patients with sickle cell anemia (SCA). The factors that affect progression of renal disease are unknown, especially in children and adolescents. Alterations in blood pressure, including hypertension and lack of the normal nocturnal dip in blood pressure, are important determinants of diabetic nephropathy and other renal diseases and may play a role in sickle cell nephropathy. Our primary hypothesis was that children with SCA who have microalbuminuria will demonstrate less nocturnal dipping of blood pressure compared to patients without microalbuminuria. We also investigated other potential factors associated with microalbuminuria. PROCEDURE: This prospective study of 52 adolescents with SCA followed in the Children's Medical Center Dallas Comprehensive Sickle Cell Center characterized 24-hour ambulatory blood pressure profiles and presence of microalbuminuria. Stepwise logistic regression was performed to identify significant independent factors that are associated with microalbuminuria. RESULTS: Thirty-five percent of patients were identified as having previously unrecognized hypertension, and 17% had pre-hypertension (blood pressure greater than the 90th percentile but less than the 95th percentile). Fifty-six percent of patients lacked the normal nocturnal dip in blood pressure. In addition, 21% had microalbuminuria, and their percent nocturnal dip was significantly less than those without microalbuminuria (P = 0.01). CONCLUSIONS: Blood pressure abnormalities are common in adolescents with SCA and are a possible modifiable risk factor in the progression of sickle cell nephropathy.

IRF-8 extinguishes neutrophil production and promotes dendritic cell lineage commitment in both myeloid and lymphoid mouse progenitors.

While most blood lineages are assumed to mature through a single cellular and developmental route downstream of HSCs, dendritic cells (DCs) can be derived from both myeloid and lymphoid progenitors in vivo. To determine how distinct progenitors can generate similar downstream lineages, we examined the transcriptional changes that accompany loss of in vivo myeloid potential as common myeloid progenitors differentiate into common DC progenitors (CDPs), and as lymphoid-primed multipotent progenitors (LMPPs) differentiate into all lymphoid progenitors (ALPs). Microarray studies revealed that IFN regulatory factor 8 (IRF-8) expression increased during each of these transitions. Competitive reconstitutions using Irf8(-/-) BM demonstrated cell-intrinsic defects in the formation of CDPs and all splenic DC subsets. Irf8(-/-) common myeloid progenitors and, unexpectedly, Irf8(-/-) ALPs produced more neutrophils in vivo than their wild-type counterparts at the expense of DCs. Retroviral expression of IRF-8 in multiple progenitors led to reduced neutrophil production and increased numbers of DCs, even in the granulocyte-macrophage progenitor (GMP), which does not normally possess conventional DC potential. These data suggest that IRF-8 represses a neutrophil module of development and promotes convergent DC development from multiple lymphoid and myeloid progenitors autonomously of cellular context.

Dynamic modulation of thymic microRNAs in response to stress.

BACKGROUND: Physiological stress evokes rapid changes in both the innate and adaptive immune response. Immature αß T cells developing in the thymus are particularly sensitive to stress, with infections and/or exposure to lipopolysaccharide or glucocorticoids eliciting a rapid apoptotic program. MicroRNAs are a class of small, non-coding RNAs that regulate global gene expression by targeting diverse mRNAs for degradation. We hypothesized that a subset of thymically encoded microRNAs would be stress responsive and modulate thymopoiesis. We performed microRNA profiling of thymic microRNAs isolated from control or stressed thymic tissue obtained from mice. We identified 18 microRNAs that are dysregulated >1.5-fold in response to lipopolysaccharide or the synthetic corticosteroid dexamethasone. These included the miR-17-90 cluster, which have anti-apoptotic functions, and the miR-181 family, which contribute to T cell tolerance. The stress-induced changes in the thymic microRNAs are dynamically and distinctly regulated in the CD4(-)CD8(-), CD4(+)CD8(+), CD4(+)CD8(-), and CD4(-)CD8(+) thymocyte subsets. Several of the differentially regulated murine thymic miRs are also stress responsive in the heart, kidney, liver, brain, and/or spleen. The most dramatic thymic microRNA down modulated is miR-181d, exhibiting a 15-fold reduction following stress. This miR has both similar and distinct gene targets as miR-181a, another member of miR-181 family. Many of the differentially regulated microRNAs have known functions in thymopoiesis, indicating that their dysregulation will alter T cell repertoire selection and the formation of naïve T cells. This data has implications for clinical treatments involving anti-inflammatory steroids, ablation therapies, and provides mechanistic insights into the consequences of infections.

Invariant NKT cell development requires a full complement of functional CD3 zeta immunoreceptor tyrosine-based activation motifs.

Invariant NKT (iNKT) cells regulate early immune responses to infections, in part because of their rapid release of IFN-gamma and IL-4. iNKT cells are proposed to reduce the severity of Lyme disease following Borrelia burgdorferi infection. Unlike conventional T cells, iNKT cells express an invariant alphabeta TCR that recognizes lipids bound to the MHC class I-like molecule, CD1d. Furthermore, these cells are positively selected following TCR interactions with glycolipid/CD1d complexes expressed on CD4+CD8+ thymocytes. Whereas conventional T cell development can proceed with as few as 4/10 CD3 immunoreceptor tyrosine-based activation motifs (ITAMs), little is known about the ITAM requirements for iNKT cell selection and expansion. We analyzed iNKT cell development in CD3 zeta transgenic lines with various tyrosine-to-phenylalanine substitutions (YF) that eliminated the functions of the first (YF1,2), third (YF5,6), or all three (YF1-6) CD3 zeta ITAMs. iNKT cell numbers were significantly reduced in the thymus, spleen, and liver of all YF mice compared with wild type mice. The reduced numbers of iNKT cells resulted from significant reductions in the expression of the early growth response 2 and promyelocytic leukemia zinc finger transcription factors. In the mice with few to no iNKT cells, there was no difference in the severity of Lyme arthritis compared with wild type controls, following infections with the spirochete B. burgdorferi. These findings indicate that a full complement of functional CD3 zeta ITAMs is required for effective iNKT cell development.

Postnatal evaluation of infants with an abnormal antenatal renal sonogram.

PURPOSE OF REVIEW: Antenatally detected renal abnormalities are frequently encountered. Recommended postnatal evaluation of these infants has evolved to minimize invasive testing while maximizing detection of significant abnormalities. RECENT FINDINGS: There is a low rate of detectable renal abnormalities in infants with a normal postnatal sonogram at 4-6 weeks of age. Routine prophylactic antibiotics are not indicated in infants with isolated antenatal hydronephrosis. Infants with a multicystic dysplastic kidney and a normal contralateral kidney on renal ultrasound do not require further evaluation. Parents of these children should be counseled on symptoms of urinary tract infections to allow prompt diagnosis. SUMMARY: All infants with abnormalities on antenatal sonogram should undergo postnatal evaluation with a sonogram after birth and at 4-6 weeks of age. Further evaluation can be safely limited when the postnatal sonogram is normal at 6 weeks of age.

Loss of E-cadherin promotes ovarian cancer metastasis via alpha 5-integrin, which is a therapeutic target.

E-cadherin loss is frequently associated with ovarian cancer metastasis. Given that adhesion to the abdominal peritoneum is the first step in ovarian cancer dissemination, we reasoned that down-regulation of E-cadherin would affect expression of cell matrix adhesion receptors. We show here that inhibition of E-cadherin in ovarian cancer cells causes up-regulation of alpha(5)-integrin protein expression and transcription. When E-cadherin was blocked, RMUG-S ovarian cancer cells were able to attach and invade more efficiently. This greater efficiency could, in turn, be inhibited both in vitro and in vivo with an alpha(5)beta(1)-integrin-blocking antibody. When E-cadherin is silenced, alpha(5)-integrin is up-regulated through activation of an epidermal growth factor receptor/FAK/Erk1-mitogen-activated protein kinase-dependent signaling pathway and not through the canonical E-cadherin/beta-catenin signaling pathway. In SKOV-3ip1 ovarian cancer xenografts, which express high levels of alpha(5)-integrin, i.p. treatment with an alpha(5)beta(1)-integrin antibody significantly reduced tumor burden, ascites, and number of metastasis and increased survival by an average of 12 days when compared with IgG treatment (P < 0.0005). alpha(5)-Integrin expression was detected by immunohistochemistry in 107 advanced stage ovarian cancers using a tissue microarray annotated with disease-specific patient follow-up. Ten of 107 tissues (9%) had alpha(5)-integrin overexpression, and 39% had some level of alpha(5)-integrin expression. The median survival for patients with high alpha(5)-integrin levels was 26 months versus 35 months for those with low integrin expression (P < 0.05). Taken together, we have identified alpha(5)-integrin up-regulation as a molecular mechanism by which E-cadherin loss promotes tumor progression, providing an explanation for how E-cadherin loss increases metastasis. Targeting this integrin could be a promising therapy for a subset of ovarian cancer patients.

Mercury intoxication: lack of correlation between symptoms and levels.

The incidence of mercury intoxication has decreased considerably because of stricter public health regulations. However, it has not been completely eliminated and should be considered in a child with unexplained tachycardia, hypertension, mood changes, weight loss, and acrodynia. Mercury intoxication can be difficult to differentiate from pheochromocytoma and Kawasaki's disease. Here, the authors report the case of an 8-year-old boy with history of mercury exposure, signs and symptoms suggestive of mercury intoxication, and good response to chelation therapy, but with only mild increase in urinary mercury levels. This case highlights the fact that urinary mercury levels do not necessarily correlate with the severity of clinical signs and symptoms of mercury intoxication.

c-Met overexpression is a prognostic factor in ovarian cancer and an effective target for inhibition of peritoneal dissemination and invasion.

The hepatocyte growth factor receptor c-Met is a receptor tyrosine kinase that plays an important role in tumor growth by activating mitogenic signaling pathways. The goal of this study was to evaluate the role of c-Met in the biology of ovarian cancer and to determine its potential as a therapeutic target. c-Met protein expression was detected by immunohistochemistry in 138 advanced-stage ovarian cancers using a tissue microarray annotated with disease-specific patient follow-up. Fifteen of 138 (11%) tissues had c-Met overexpression. Median survival for patients with high c-Met levels was 17 months versus 32 months (P = 0.001) for patients with low c-Met expression. Infection of SKOV-3ip1 cells with an adenovirus expressing a small interfering RNA (siRNA) against c-Met efficiently inhibited c-Met protein and mRNA expression as well as extracellular signal-regulated kinase and phosphatidylinositol 3-kinase signaling. It also inhibited adhesion to different extracellular matrix components, human primary mesothelial cells, and full-thickness human peritoneum and, in vivo, to mouse peritoneum. This was paralleled by a significant reduction in alpha(5) and beta(1) integrin protein and mRNA expression as well as a reduction of urokinase and matrix metalloproteinase (MMP)-2/MMP-9 activity. In SKOV-3ip1 ovarian cancer xenografts, i.p. treatment with the c-Met siRNA significantly reduced tumor burden, ascites formation, protease activity, and the number of peritoneal implants but not tumor size or angiogenesis. These results suggest that c-Met overexpression is a prognostic factor in ovarian cancer and that targeting c-Met in vivo inhibits peritoneal dissemination and invasion through an alpha(5)beta(1) integrin-dependent mechanism. Therefore, c-Met should be explored further as a therapeutic target in ovarian cancer.

Primary renal lymphoma presenting with hypertension.

Primary renal lymphoma (PRL) is a rare lymphoma which usually presents with hematuria, flank pain, abdominal mass, and weight loss. PRL is more diagnosed in adults than children. We describe an asymptomatic child who presented with hypertension and was subsequently diagnosed with primary renal lymphoma. This case represents an atypical presentation for PRL.

Obstructive uropathy.

Routine second trimester ultrasound screening has resulted in more infants diagnosed with antenatal hydronephrosis. Current recommendations suggest postnatal evaluation of all infants with a renal pelvic diameter >5 mm with ultrasound and voiding cystourethrogram (VCUG.) There are many etiologies of obstructive uropathy including ureteropelvic junction (UPJ) obstruction, ureterovesical junction (UVJ) obstruction, posterior urethral valves (PUV), prune belly syndrome, and vesicoureteral reflux (VUR). Obstructive uropathy can result in tubular damage and decreased nephron number. Tubular damage can result in sodium wasting, hyperkalemic acidosis, and nephrogenic diabetes insipidus. Most patients do not require renal replacement therapy in the neonatal period; however, chronic renal insufficiency can occur if the neonate has a significant reduction in nephron number or progressive renal damage from obstruction or infection.