Gefitinib targets ZAP-70-expressing chronic lymphocytic leukemia cells and inhibits B-cell receptor signaling.

Chronic lymphocytic leukemia (CLL) can be divided into groups based on biomarkers of poor prognosis. The expression of the tyrosine kinase ZAP-70 (member of the Syk tyrosine kinase family) in CLL cells is associated with shorter overall survival in CLL patients. Currently, there is a lack of targeted therapies for patients with ZAP-70 expression in CLL cells. The tyrosine kinase inhibitor gefitinib has been shown to be effective at induce apoptosis in acute myeloid leukemia through inhibition of Syk. In this study, we sought to test the efficacy of gefitinib in primary human ZAP-70+ CLL cells. We demonstrate that gefitinib preferentially induces cell death in ZAP-70-expressing CLL cells with a median IC50 of 4.5 µM. In addition, gefitinib decreases the viability of ZAP-70+ Jurkat T leukemia cells but fails to affect T cells from CLL patients. Western blot analysis shows gefitinib reduces both basal and B-cell receptor (BCR)-stimulated phosphorylation of Syk/ZAP-70, ERK, and Akt in ZAP-70+ CLL cells. Moreover, gefitinib inhibits the pro-survival response from BCR stimulation and decreases pro-survival proteins such as Mcl-1. Finally, ZAP-70 expression sensitizes Raji cells to gefitinib treatment. These results demonstrate that gefitinib specifically targets ZAP-70+ CLL cells and inhibits the BCR cell survival pathway leading to apoptosis. This represents the likelihood of tyrosine kinase inhibitors being effective targeted treatments for ZAP-70+ CLL cells.

Follicular dendritic cell secreted protein FDC-SP controls IgA production.

Follicular dendritic cell secreted protein (FDC-SP) is a secreted peptide predominantly expressed in mucosal tissues. We previously reported that FDC-SP transgenic mice have altered B-cell responses to systemic immunization; however, the role of FDC-SP in mucosal immunity is unknown. Here, we report that FDC-SP functions in regulating immunoglobulin A production. FDC-SP transgenic mice show decreased IgA levels in serum, saliva, and bronchoalveolar lavage fluid. Reciprocally, FDC-SP-deficient mice show significantly increased IgA levels in serum and intestinal lavage, associated with accumulation of IgA+ cells in blood, bone marrow, Peyer's patches, and lymph nodes. FDC-SP-deficient mice generated higher titers of antigen-specific IgA but normal IgG1 responses upon immunization. Purified FDC-SP transgenic B cells generated decreased IgA responses to transforming growth factor ß (TGFß)+interleukin 5 (IL5) stimulation. Consistent with a direct effect of FDC-SP on B cells, recombinant FDC-SP suppressed B-cell IgA production in vitro. Six- to 14-month-old FDC-SP-deficient mice show IgA deposition in kidney glomeruli, which was associated with proteinuria and pathology consistent with mild IgA nephropathy (IgAN). Our results demonstrate a novel biological activity of FDC-SP in controlling B-cell IgA production and identify FDC-SP-deficient mice as a novel mouse model of IgAN.

Epidermal and dermal integration into sphere-templated porous poly(2-hydroxyethyl methacrylate) implants in mice.

Percutaneous medical devices remain susceptible to infection and failure. We hypothesize that healing of the skin into the percutaneous device will provide a seal, preventing bacterial attachment, biofilm formation, and subsequent device failure. Porous poly(2-hydroxyethyl methacrylate) [poly(HEMA)] with sphere-templated pores (40 microm) and interconnecting throats (16 microm) were implanted in normal C57BL/6 mice for 7, 14, and 28 days. Poly(HEMA) was either untreated, keeping the surface nonadhesive for cells and proteins, or modified with carbonyldiimidazole (CDI) or CDI reacted with laminin 332 to enhance adhesion. No clinical signs of infection were observed. Epidermal and dermal response within the poly(HEMA) pores was evaluated using light and transmission electron microscopy. Cells (keratinocytes, fibroblasts, endothelial cells, inflammatory cells) and basement membrane proteins (laminin 332, beta4 integrin, type VII collagen) could be demonstrated within the poly(HEMA) pores of all implants. Blood vessels and dermal collagen bundles were evident in all of the 14- and 28-day implants. Fibrous capsule formation and permigration were not observed. Sphere-templated polymers with 40 microm pores demonstrate an ability to recapitulate key elements of both the dermal and the epidermal layers of skin. Our morphological findings indicate that the implant model can be used to study the effects of biomaterial pore size, pore interconnect (throat) size, and surface treatments on cutaneous biointegration. Further, this model may be used for bacterial challenge studies.

Evaluating orangutan census techniques using nest decay rates: implications for population estimates.

An accurate estimate for orangutan nest decay time is a crucial factor in commonly used methods for estimating orangutan population size. Decay rates are known to vary, but the decay process and, thus, the temporal and spatial variation in decay time are poorly understood. We used established line-transect methodology to survey orangutan nests in a lowland forest in East Kalimantan, Indonesia, and monitored the decay of 663 nests over 20 months. Using Markov chain analysis we calculated a decay time of 602 days, which is significantly longer than times found in other studies. Based on this, we recalculated the orangutan density estimate for a site in East Kalimantan; the resulting density is much lower than previous estimates (previous estimates were 3-8 times higher than our recalculated density). Our data suggest that short-term studies where decay times are determined using matrix mathematics may produce unreliable decay times. Our findings have implications for other parts of the orangutan range where population estimates are based on potentially unreliable nest decay rate estimates, and we recommend that for various parts of the orangutan range census estimates be reexamined. Considering the high variation in decay rates there is a need to move away from using single-number decay time estimates and, preferably, to test methods that do not rely on nest decay times as alternatives for rapid assessments of orangutan habitat for conservation in Borneo.

Impaired left ventricular function has a detrimental effect on image quality in multi-detector row CT coronary angiography.

AIM: To determine whether there is a relationship between left ventricular (LV) haemodynamic parameters, circulation times, and arterial contrast opacification that might affect the image quality of computed tomography (CT) coronary angiography. METHODS: Thirty-six patients were included in the study: 18 with cardiomyopathy (CM) and LV dilatation of suspected ischaemic aetiology [age 57.9+/-13.7 years, range 30-77 years; 14 male, four female; body mass index (BMI)=27.7+/-4.5, range 25.5-31.8] and 18 controls (age 62.3+/-9.4 years, range 47-89 years; 10 male, eight female; BMI 27.8+/-6.6; range 19.2-33.6). Coronary artery image quality was assessed using a three-point visual scale; contrast medium circulation times, aortic root contrast attenuation, and LV functional parameters were studied. RESULTS: Visually reduced contrast opacification impaired image quality more often in the CM group than the control group (27.4 versus 5.1%). A total of 55.6% CM patients had a contrast transit time ranging from 30-75 s; the number of "unassessable" segments increased with increasing transit time conforming to a fitted quadratic model (R2=0.74). The relationship between LV ejection fraction and contrast attenuation may also conform to a quadratic model (R2=0.71). CONCLUSION: LV haemodynamics influence coronary artery opacification using cardiac CT, and users imaging this subgroup must do so with the knowledge of this potential pitfall. The results indicate the need for further studies examining CT protocols in this clinical subgroup.

A mouse model to evaluate the interface between skin and a percutaneous device.

Percutaneous medical devices are integral in the management and treatment of disease. The space created between the skin and the device becomes a haven for bacterial invasion and biofilm formation and results in infection. We hypothesize that sealing this space via integration of the skin into the device will create a barrier against bacterial invasion. The purpose of this study was to develop an animal model in which the interaction between skin and biomaterials can be evaluated. Porous poly(2-hydroxyethyl methacrylate) [poly(HEMA)] rods were implanted for 7 days in the dorsal skin of C57 BL/6 mice. The porous poly(HEMA) rods were surface-modified with carbonyldiimidazole (CDI) or CDI plus laminin 5; unmodified rods served as control. Implant sites were sealed with 2-octyl cyanoacrylate; corn pads and adhesive dressings were tested for stabilization of implants. All rods remained intact for the duration of the study. There was histological evidence of both epidermal and dermal integration into all poly(HEMA) rods regardless of treatment. This in vivo model permits examination of the implant/skin interface and will be useful for future studies designed to facilitate skin cell attachment where percutaneous devices penetrate the skin.

Multi-detector row CT coronary angiography in patients with cardiomyopathy -- initial single-centre experience.

AIMS: To evaluate the diagnostic accuracy of computed tomography (CT) in assessing haemodynamically significant coronary artery stenoses in patients with cardiomyopathy (CM). SUBJECTS AND METHODS: Eighteen patients with CM were approached to undergo CT coronary angiography to evaluate the use of this technique for investigating the presence of significant coronary artery disease (CAD), and also to compare the findings with catheter angiography. RESULTS: On a segment-by-segment analysis the sensitivity, specificity, positive and negative predictive values in the CM group were 66.7, 96.5, 40 and 98.8%, respectively, with 100% accuracy in "whole-patient terms". CONCLUSION: Non-invasive, 16-detector row CT coronary angiography in patients with presumed CM would seem to be a useful clinical tool for the exclusion of significant coronary artery disease. However, the presence of suboptimal contrast opacification in this patient group means that the implication of these results must be interpreted with caution.

Regulation of B-lymphocyte activation by the PH domain adaptor protein Bam32/DAPP1.

PI3Ks (phosphoinositide 3-kinases) play critical roles in BCR (B-cell receptor) signalling via the generation of 3-phosphoinositide second messengers. Recruitment of PH domain (pleckstrin homology domain)-containing signal transduction proteins to the plasma membrane through binding to 3-phosphoinositide second messengers represents a major effector mechanism for PI3Ks. Here, we review data on the PH domain-containing adaptor protein Bam32 (B-cell adaptor molecule of 32 kDa)/DAPP1 (dual adaptor for phosphotyrosine and 3-phosphoinositides 1), focusing on its functions in B-lymphocyte activation. Present results support the view that Bam32/DAPP1 mediates multiple PI3K-dependent responses in B-cells through membrane-proximal mechanisms involving Src kinases, Rac1, F-actin and mitogen-activated protein kinases, resulting in selective effects on BCR-mediated proliferation, antigen presentation and generation of antibody responses.

16-detector row computed tomographic coronary angiography in patients undergoing evaluation for aortic valve replacement: comparison with catheter angiography.

AIM: To evaluate the diagnostic accuracy of 16-detector row computed tomography (CT) in assessing haemodynamically significant coronary artery stenoses in patients under evaluation for aortic stenosis pre-aortic valve replacement. SUBJECTS AND METHODS: Forty consecutive patients under evaluation for severe aortic stenosis and listed for cardiac catheterization before potential aortic valve replacement underwent coronary artery calcium (CAC) scoring and retrospective electrocardiogram (ECG)-gated multi-detector row computed tomographic coronary angiography (MDCTA) using a GE Lightspeed 16-detector row CT within 1 month of invasive coronary angiography (ICA) for comparative purposes. All 13 major coronary artery segments of the American Heart Association model were evaluated for the presence of > or =50% stenosis and compared to the reference standard. Data were analysed on a segment-by-segment basis and also in "whole patient" terms. RESULTS: A total of 412/450 segments from 35 patients were suitable for analysis. The overall accuracy of MDCTA for detection of segments with > or =50% stenosis was high, with a sensitivity of 81.3%, specificity 95.0%, positive predictive value (PPV) 57.8%, and negative predictive value (NPV) 98.4%. On a "whole-patient" basis, 100% (19/19) of patients with significant coronary disease were correctly identified and there were no false-negatives. Excluding patients with CAC >1000 from the analysis improved the accuracy of MDCTA to: sensitivity 90%, specificity 98.1%, PPV 60%, NPV 99.7%. CONCLUSION: Non-invasive 16-detector row MDCTA accurately excludes significant coronary disease in patients with severe aortic stenosis undergoing evaluation before aortic valve replacement and in whom ICA can therefore be avoided. Its segment-by-segment accuracy is improved further if CAC>1000 is used as a gatekeeper to MDCTA.

Multidetector row computed tomography: imaging congenital coronary artery anomalies in adults.

The quality of the imaging of the main coronary arteries and side branches provided by multidetector row computed tomography (MDCT) may have importance when assessing congenital coronary artery anomalies. This review discusses the rationale for using MDCT for this indication and examines the advantages and disadvantages of the technique. Examples of MDCT imaging of congenital coronary artery anomalies are presented. These images provide persuasive evidence to support clinical use of MDCT cardiac imaging in the context of suspected congenital coronary artery anomalies as a first line investigation.

Multi-detector row computed tomography: imaging the coronary arteries.

Over the last 2 years, multi-detector row computed tomographic (MDCT) cardiac imaging has continued to rapidly develop and evolve from the experimental research setting to become a useful clinical tool. The increasing availability of MDCT presents today's clinicians with an additional non-invasive diagnostic cardiac imaging method, in particular for the coronary arteries. With the advent and increasing clinical use of 16-detector row machines, and now with the imminent clinical emergence of 64-channel machines, the improvements in spatial and temporal resolution and sophisticated ECG-gating are allowing motion-free, fast, accurate, detailed, contrast-enhanced cardiac imaging that begins to approach the accuracy of traditional invasive diagnostic techniques. Additional diagnostic information may also be provided.

Highly accurate coronary angiography with submillimetre, 16 slice computed tomography.

OBJECTIVE: To assess submillimetre coronary computed tomographic angiography (CTA) in comparison with invasive quantitative coronary angiography as the gold standard and to examine the effect of significant coronary artery calcification (CAC), which is known to impede lumen visualisation, on the accuracy of the examination. METHODS: After invasive coronary angiography, 58 patients underwent coronary imaging with a GE Lightspeed 16 computed tomography (CT) system. CAC was quantified after an ECG triggered acquisition with a low tube current. Coronary CTA was performed with retrospective ECG gating and a 16 x 0.63 mm collimation and was reconstructed with an effective 65-250 ms temporal resolution. All 13 major coronary artery segments were evaluated for the presence of > or = 50% stenosis, and compared with the gold standard. RESULTS: One patient moved and could not be evaluated. All segments (except occluded segments) were evaluated for 57 patients. Overall the accuracy of coronary CTA for detection of > or = 50% stenosis was: sensitivity 83%, specificity 97%, positive predictive value 80%, and negative predictive value 97%. The number of diseased coronary arteries was correctly diagnosed in 34 of 38 (89%) patients overall. Altogether 21 of 57 (37%) patients had a CAC score > or = 400, which was predefined as representing significant CAC. Excluding these patients from the analysis improved the accuracy of coronary CTA to a sensitivity of 89%, specificity 98%, positive predictive value 79%, and negative predictive value 99%. CONCLUSIONS: Non-invasive coronary angiography with submillimetre CT is reliable and accurate. It appears that a subgroup of patients may be selected based on CAC score in whom the investigation has even higher accuracy. Coronary CTA has reached the stage where it should be considered for a clinical role. Further research is required to define this role.

Glucose-sensitive holographic sensors.

Holographic sensors for monitoring glucose were fabricated from hydrogel films containing chemical ligands based on phenylboronic acid. The films were transformed into reflection holograms using a diffusion method coupled with exposure to laser light. The diffraction wavelength of the holograms was used to monitor the swelling of the hydrogel film in the presence of glucose. Fully reversible changes in diffraction wavelength were demonstrated, highlighting the potential for using these holograms as glucose sensors.

Three dimensional volume quantification of aortic valve calcification using multislice computed tomography.

OBJECTIVE: To assess a new multislice computed tomography (CT) technique for three dimensional quantification of aortic valve calcification volume (3D AVCV) and to study the relation between stenosis and calcification of the aortic valve. METHODS: 50 patients with echocardiographic calcification of the aortic valve underwent two separate ECG triggered multislice CT for quantification of 3D AVCV. The agreement between the two 3D AVCV scores was assessed and 3D AVCV was compared with echocardiographic markers of severity of aortic stenosis. RESULTS: Overall the level of agreement between the two 3D AVCV scores was excellent (median interscan variability 7.9% (interquartile range 10.1); correlation coefficient, r = 0.99; repeatability coefficient 237.8 mm3 (limits of agreement -393 to 559 mm3)). However, the magnitude of the 3D AVCV did influence the interscan variability. The 3D AVCV correlated closely with the maximal predicted transvalvar gradient (r2 = 0.77) and aortic valve area (r2 = 0.73). CONCLUSIONS: Multislice CT provides a technique for quantifying 3D AVCV that has good reproducibility. There is a close non-linear relation between echocardiographic parameters of severity of valve stenosis and 3D AVCV scores.

Refined computed tomography of the thoracic aorta: the impact of electrocardiographic assistance.

There have been a number of advances in helical computed tomography (CT) in recent years, which have had a beneficial impact on the quality of imaging of the thoracic aorta. These advances include sub-second gantry rotation, multislice acquisition, and the use of electrocardiographic (ECG) assistance. We examine these techniques with emphasis on the principles behind ECG assistance and its use to reduce aortic motion artefact. We highlight examples of ECG-assisted multislice CT in a spectrum of pathologies of the thoracic aorta.