Severity mapping of the proximal femur: a new method for assessing hip osteoarthritis with computed tomography.

OBJECTIVE: Plain radiography has been the mainstay of imaging assessment in osteoarthritis for over 50 years, but it does have limitations. Here we present the methodology and results of a new technique for identifying, grading, and mapping the severity and spatial distribution of osteoarthritic disease features at the hip in 3D with clinical computed tomography (CT). DESIGN: CT imaging of 456 hips from 230 adult female volunteers (mean age 66 ± 17 years) was reviewed using 3D multiplanar reformatting to identify bone-related radiological features of osteoarthritis, namely osteophytes, subchondral cysts and joint space narrowing. Scoresheets dividing up the femoral head, head-neck region and the joint space were used to register the location and severity of each feature (scored from 0 to 3). Novel 3D cumulative feature severity maps were then created to display where the most severe disease features from each individual were anatomically located across the cohort. RESULTS: Feature severity maps showed a propensity for osteophytes at the inferoposterior and superolateral femoral head-neck junction. Subchondral cysts were a less common and less localised phenomenon. Joint space narrowing <1.5 mm was recorded in at least one sector of 83% of hips, but most frequently in the posterolateral joint space. CONCLUSIONS: This is the first description of hip osteoarthritis using unenhanced clinical CT in which we describe the co-localisation of posterior osteophytes and joint space narrowing for the first time. We believe this technique can perform several important roles in future osteoarthritis research, including phenotyping and sensitive disease assessment in 3D.KP acknowledges support of an Arthritis Research UK Research Progression award, and the Cambridge NIHR Biomedical Research Centre (MEBB theme). TT acknowledges the support of an Evelyn Trust Clinical Training Fellowship award. None of the funding sources had a role in study design, data handling, writing of the report, or decision to submit the paper for publication.This is the final version. It was first published by Elsevier at http://www.sciencedirect.com/science/article/pii/S1063458414009996?np=

A new CT grading system for hip osteoarthritis.

OBJECTIVES: We have developed a new grading system for hip osteoarthritis using clinical computed tomography (CT). This technique was compared with Kellgren and Lawrence (K&L) grading and minimum joint space width (JSW) measurement in digitally reconstructed radiographs (DRRs) from the same CT data. In this paper we evaluate and compare the accuracy and reliability of these measures in the assessment of radiological disease. DESIGN: CT imaging of hips from 30 female volunteers aged 66 ± 17 years were used in two reproducibility studies, one testing the reliability of the new system, the other testing K&L grading and minimum JSW measurement in DRRs. RESULTS: Intra- and inter-observer reliability was substantial for CT grading according to weighted kappa (0.74 and 0.75 respectively), while intra- and inter-observer reliability was at worst moderate (0.57) and substantial (0.63) respectively for DRR K&L grading. Bland-Altman analysis showed a systematic difference in minimum JSW measurement of 0.82 mm between reviewers, with a least detectable difference of 1.06 mm. The area under the curve from ROC analysis was 0.91 for our CT composite score. CONCLUSIONS: CT grading of hip osteoarthritis (categorised as none, developing and established) has substantial reliability. Sensitivity was increased when CT features of osteoarthritis were assigned a composite score (0 = none to 7 = severest) that also performed well as a diagnostic test, but at the cost of reliability. Having established feasibility and reliability for this new CT system, sensitivity testing and validation against clinical measures of hip osteoarthritis will now be performed.KP acknowledges support of an Arthritis Research UK Research Progression award, and the Cambridge NIHR Biomedical Research Centre (MEBB theme). TT acknowledges the support of an Evelyn Trust Clinical Training Fellowship award. None of the funding sources had a role in study design, data handling, writing of the report, or decision to submit the paper for publication.This is the final version. It was first published by Elsevier at http://www.sciencedirect.com/science/article/pii/S106345841401000

Mass spectrometry of B. subtilis CopZ: Cu(I)-binding and interactions with bacillithiol

CopZ from Bacillus subtilis is a well-studied member of the highly conserved family of Atx1-like copper chaperones. It was previously shown via solution and crystallographic studies to undergo Cu(I)-mediated dimerisation, where the CopZ dimer can bind between one and four Cu(I) ions. However, these studies could not provide information about the changing distribution of species at increasing Cu(I) levels. To address this, electrospray ionisation mass spectrometry using soft ionisation was applied to CopZ under native conditions. Data revealed folded, monomeric CopZ in apo- and Cu(I)-bound forms, along with Cu(I)-bound dimeric forms of CopZ at higher Cu(I) loading. Cu4(CopZ)2 was the major dimeric species at loadings >1 Cu(I)/CopZ, indicating the cooperative formation of the tetranuclear Cu(I)-bound species. As the principal low molecular weight thiol in B. subtilis, bacillithiol (BSH) may play a role in copper homeostasis. Mass spectrometry showed that increasing BSH led to a reduction in Cu(I)-bound dimeric forms, and the formation of S-bacillithiolated apo-CopZ and BSH adducts of Cu(I)-bound forms of CopZ, where BSH likely acts as a Cu(I) ligand. These data, along with the high affinity of BSH for Cu(I), determined here to be β2(BSH) = ∼4 × 1017 M−2, are consistent with a role for BSH alongside CopZ in buffering cellular Cu(I) levels. Here, mass spectrometry provides a high resolution overview of CopZ–Cu(I) speciation that cannot be obtained from less discriminating solution-phase methods, thus illustrating the potential for the wider application of this technique to studies of metal–protein interactions

Tetranucleotide and Low Microsatellite Instability Are Inversely Associated with the CpG Island Methylator Phenotype in Colorectal Cancer

MSH3 gene or protein deficiency or loss-of-function in colorectal cancer can cause a DNA mismatch repair defect known as “elevated microsatellite alterations at selected tetranucleotide repeats” (EMAST). A high percentage of MSI-H tumors exhibit EMAST, while MSI-L is also linked with EMAST. However, the distribution of CpG island methylator phenotype (CIMP) within the EMAST spectrum is not known. Five tetranucleotide repeat and five MSI markers were used to classify 100 sporadic colorectal tumours for EMAST, MSI-H and MSI-L according to the number of unstable markers detected. Promoter methylation was determined using methylation-specific PCR for MSH3, MCC, CDKN2A (p16) and five CIMP marker genes. EMAST was found in 55% of sporadic colorectal carcinomas. Carcinomas with only one positive marker (EMAST-1/5, 26%) were associated with advanced tumour stage, increased lymph node metastasis, MSI-L and lack of CIMP-H. EMAST-2/5 (16%) carcinomas displayed some methylation but MSI was rare. Carcinomas with ≥3 positive EMAST markers (13%) were more likely to have a proximal colon location and be MSI-H and CIMP-H. Our study suggests that EMAST/MSI-L is a valuable prognostic and predictive marker for colorectal carcinomas that do not display the high methylation phenotype CIMP-H

Colorectal cancer linkage on chromosomes 4q21, 8q13, 12q24, and 15q22

A substantial proportion of familial colorectal cancer (CRC) is not a consequence of known susceptibility loci, such as mismatch repair (MMR) genes, supporting the existence of additional loci. To identify novel CRC loci, we conducted a genome-wide linkage scan in 356 white families with no evidence of defective MMR (i.e., no loss of tumor expression of MMR proteins, no microsatellite instability (MSI)-high tumors, or no evidence of linkage to MMR genes). Families were ascertained via the Colon Cancer Family Registry multi-site NCI-supported consortium (Colon CFR), the City of Hope Comprehensive Cancer Center, and Memorial University of Newfoundland. A total of 1,612 individuals (average 5.0 per family including 2.2 affected) were genotyped using genome-wide single nucleotide polymorphism linkage arrays; parametric and non-parametric linkage analysis used MERLIN in a priori-defined family groups. Five lod scores greater than 3.0 were observed assuming heterogeneity. The greatest were among families with mean age of diagnosis less than 50 years at 4q21.1 (dominant HLOD = 4.51, α = 0.84, 145.40 cM, rs10518142) and among all families at 12q24.32 (dominant HLOD = 3.60, α = 0.48, 285.15 cM, rs952093). Among families with four or more affected individuals and among clinic-based families, a common peak was observed at 15q22.31 (101.40 cM, rs1477798; dominant HLOD = 3.07, α = 0.29; dominant HLOD = 3.03, α = 0.32, respectively). Analysis of families with only two affected individuals yielded a peak at 8q13.2 (recessive HLOD = 3.02, α = 0.51, 132.52 cM, rs1319036). These previously unreported linkage peaks demonstrate the continued utility of family-based data in complex traits and suggest that new CRC risk alleles remain to be elucidated. © 2012 Cicek et al

Clinical evaluation of smartphone-based fluorescence imaging for guidance and monitoring of ALA PDT

India has one of the highest rates of oral cancer incidence in the world, with an estimated 80,000 new cases per year, accounting for 30% of reported cancers. In rural areas, a lack of adequate medical infrastructure contributes to unchecked disease progression and dismal mortality rates. PDT emerges as a potential modality which can be implemented in resource limited settings, while photosensitizer fluorescence can be leveraged for treatment guidance. Here, as part of an ongoing clinical study evaluating low-cost technology for ALA PDT treatment, we evaluated the capability of a simple smartphone-based device for imaging ALA-induced PpIX fluorescence. The imaging device itself consists of an annulus of 405nm LEDs for PpIX excitation with emission filter in the center mounted over the phone camera. 18 subjects having <2 cm diameter (mean size; ~1.38 cm2) lesions with micro-invasive (≤5 mm. depth) moderately/well-differentiated squamous cell carcinoma were administered 60 mg/kg ALA in oral solution and imaged before and after delivery of 100 J/cm2 total light dose to the lesion surface. We will present comparative analysis of pre-and post-treatment fluorescence, white light, and ultrasound images. In general, PpIX fluorescence images obtained prior to therapeutic light delivery are able to resolve lesion margins while dramatic photobleaching in post-treatment images confirms the irradiated zone. Overall this approach is able to generate sufficient fluorescence contrast for treatment guidance and monitoring photobleaching while the use of a smartphone-based device provides a low-cost, widely available platform with potential for telemedicine integration