The zebrafish kidney mutant zeppelin reveals that brca2/fancd1 is essential for pronephros development.

The zebrafish kidney is conserved with other vertebrates, making it an excellent genetic model to study renal development. The kidney collects metabolic waste using a blood filter with specialized epithelial cells known as podocytes. Podocyte formation is poorly understood but relevant to many kidney diseases, as podocyte injury leads to progressive scarring and organ failure. zeppelin (zep) was isolated in a forward screen for kidney mutants and identified as a homozygous recessive lethal allele that causes reduced podocyte numbers, deficient filtration, and fluid imbalance. Interestingly, zep mutants had a larger interrenal gland, the teleostean counterpart of the mammalian adrenal gland, which suggested a fate switch with the related podocyte lineage since cell proliferation and cell death were unchanged within the shared progenitor field from which these two identities arise. Cloning of zep by whole genome sequencing (WGS) identified a splicing mutation in breast cancer 2, early onset (brca2)/fancd1, which was confirmed by sequencing of individual fish. Several independent brca2 morpholinos (MOs) phenocopied zep, causing edema, reduced podocyte number, and increased interrenal cell number. Complementation analysis between zep and brca2ZM_00057434 -/- zebrafish, which have an insertional mutation, revealed that the interrenal lineage was expanded. Importantly, overexpression of brca2 rescued podocyte formation in zep mutants, providing critical evidence that the brca2 lesion encoded by zep specifically disrupts the balance of nephrogenesis. Taken together, these data suggest for the first time that brca2/fancd1 is essential for vertebrate kidney ontogeny. Thus, our findings impart novel insights into the genetic components that impact renal development, and because BRCA2/FANCD1 mutations in humans cause Fanconi anemia and several common cancers, this work has identified a new zebrafish model to further study brca2/fancd1 in disease.

Image intensifier distortion correction for fluoroscopic RSA: the need for independent accuracy assessment.

Fluoroscopic images suffer from multiple modes of image distortion. Therefore, the purpose of this study was to compare the effects of correction using a range of two-dimensional polynomials and a global approach. The primary measure of interest was the average error in the distances between four beads of an accuracy phantom, as measured using RSA. Secondary measures of interest were the root mean squared errors of the fit of the chosen polynomial to the grid of beads used for correction, and the errors in the corrected distances between the points of the grid in a second position. Based upon the two-dimensional measures, a polynomial of order three in the axis of correction and two in the perpendicular axis was preferred. However, based upon the RSA reconstruction, a polynomial of order three in the axis of correction and one in the perpendicular axis was preferred. The use of a calibration frame for these three-dimensional applications most likely tempers the effects of distortion. This study suggests that distortion correction should be validated for each of its applications with an independent "gold standard" phantom.

The effect of decreasing computed tomography dosage on radiostereometric analysis (RSA) accuracy at the glenohumeral joint.

Standard, beaded radiostereometric analysis (RSA) and markerless RSA often use computed tomography (CT) scans to create three-dimensional (3D) bone models. However, ethical concerns exist due to risks associated with CT radiation exposure. Therefore, the aim of this study was to investigate the effect of decreasing CT dosage on RSA accuracy. Four cadaveric shoulder specimens were scanned using a normal-dose CT protocol and two low-dose protocols, where the dosage was decreased by 89% and 98%. 3D computer models of the humerus and scapula were created using each CT protocol. Bi-planar fluoroscopy was used to image five different static glenohumeral positions and two dynamic glenohumeral movements, of which a total of five static and four dynamic poses were selected for analysis. For standard RSA, negligible differences were found in bead (0.21±0.31mm) and bony landmark (2.31±1.90mm) locations when the CT dosage was decreased by 98% (p-values>0.167). For markerless RSA kinematic results, excellent agreement was found between the normal-dose and lowest-dose protocol, with all Spearman rank correlation coefficients greater than 0.95. Average root mean squared errors of 1.04±0.68mm and 2.42±0.81° were also found at this reduced dosage for static positions. In summary, CT dosage can be markedly reduced when performing shoulder RSA to minimize the risks of radiation exposure. Standard RSA accuracy was negligibly affected by the 98% CT dose reduction and for markerless RSA, the benefits of decreasing CT dosage to the subject outweigh the introduced errors.

The effect of CT dose on glenohumeral joint congruency measurements using 3D reconstructed patient-specific bone models.

The study of joint congruency at the glenohumeral joint of the shoulder using computed tomography (CT) and three-dimensional (3D) reconstructions of joint surfaces is an area of significant clinical interest. However, ionizing radiation delivered to patients during CT examinations is much higher than other types of radiological imaging. The shoulder represents a significant challenge for this modality as it is adjacent to the thyroid gland and breast tissue. The objective of this study was to determine the optimal CT scanning techniques that would minimize radiation dose while accurately quantifying joint congruency of the shoulder. The results suggest that only one-tenth of the standard applied total current (mA) and a pitch ratio of 1.375:1 was necessary to produce joint congruency values consistent with that of the higher dose scans. Using the CT scanning techniques examined in this study, the effective dose applied to the shoulder to quantify joint congruency was reduced by 88.9% compared to standard clinical CT imaging techniques.