Temporal and Spatial Uncoupling of DNA Double Strand Break Repair Pathways within Mammalian Heterochromatin.

Repetitive DNA is packaged into heterochromatin to maintain its integrity. We use CRISPR/Cas9 to induce DSBs in different mammalian heterochromatin structures. We demonstrate that in pericentric heterochromatin, DSBs are positionally stable in G1 and recruit NHEJ factors. In S/G2, DSBs are resected and relocate to the periphery of heterochromatin, where they are retained by RAD51. This is independent of chromatin relaxation but requires end resection and RAD51 exclusion from the core. DSBs that fail to relocate are engaged by NHEJ or SSA proteins. We propose that the spatial disconnection between end resection and RAD51 binding prevents the activation of mutagenic pathways and illegitimate recombination. Interestingly, in centromeric heterochromatin, DSBs recruit both NHEJ and HR proteins throughout the cell cycle. Our results highlight striking differences in the recruitment of DNA repair factors between pericentric and centromeric heterochromatin and suggest a model in which the commitment to specific DNA repair pathways regulates DSB position.

Cell-based cccDNA reporter assay combined with functional genomics identifies YBX1 as HBV cccDNA host factor and antiviral candidate target.

OBJECTIVES: Chronic hepatitis B virus (HBV) infection is a leading cause of liver disease and hepatocellular carcinoma. A key feature of HBV replication is the synthesis of the covalently close circular (ccc)DNA, not targeted by current treatments and whose elimination would be crucial for viral cure. To date, little is known about cccDNA formation. One major challenge to address this urgent question is the absence of robust models for the study of cccDNA biology. DESIGN: We established a cell-based HBV cccDNA reporter assay and performed a loss-of-function screen targeting 239 genes encoding the human DNA damage response machinery. RESULTS: Overcoming the limitations of current models, the reporter assay enables to quantity cccDNA levels using a robust ELISA as a readout. A loss-of-function screen identified 27 candidate cccDNA host factors, including Y box binding protein 1 (YBX1), a DNA binding protein regulating transcription and translation. Validation studies in authentic infection models revealed a robust decrease in HBV cccDNA levels following silencing, providing proof-of-concept for the importance of YBX1 in the early steps of the HBV life cycle. In patients, YBX1 expression robustly correlates with both HBV load and liver disease progression. CONCLUSION: Our cell-based reporter assay enables the discovery of HBV cccDNA host factors including YBX1 and is suitable for the characterisation of cccDNA-related host factors, antiviral targets and compounds.

ARCH domain of XPD, an anchoring platform for CAK that conditions TFIIH DNA repair and transcription activities.

The xeroderma pigmentosum group D (XPD) helicase is a subunit of transcription/DNA repair factor, transcription factor II H (TFIIH) that catalyzes the unwinding of a damaged DNA duplex during nucleotide excision repair. Apart from two canonical helicase domains, XPD is composed of a 4Fe-S cluster domain involved in DNA damage recognition and a module of uncharacterized function termed the "ARCH domain." By investigating the consequences of a mutation found in a patient with trichothiodystrophy, we show that the ARCH domain is critical for the recruitment of the cyclin-dependent kinase (CDK)-activating kinase (CAK) complex. Indeed, this mutation not only affects the interaction with the MAT1 CAK subunit, thereby decreasing the in vitro basal transcription activity of TFIIH itself and impeding the efficient recruitment of the transcription machinery on the promoter of an activated gene, but also impairs the DNA unwinding activity of XPD and the nucleotide excision repair activity of TFIIH. We further demonstrate the role of CAK in downregulating the XPD helicase activity within TFIIH. Taken together, our results identify the ARCH domain of XPD as a platform for the recruitment of CAK and as a potential molecular switch that might control TFIIH composition and play a key role in the conversion of TFIIH from a factor active in transcription to a factor involved in DNA repair.

3.0 Tesla breast magnetic resonance imaging in patients with nipple discharge when mammography and ultrasound fail.

OBJECTIVES: To compare 3.0 Tesla breast magnetic resonance imaging (MRI) with galactography for detection of benign and malignant causes of nipple discharge in patients with negative mammography and ultrasound. METHODS: We prospectively evaluated 56 breasts of 50 consecutive patients with nipple discharge who had inconspicuous mammography and ultrasound, using 3.0 Tesla breast MRI with a dedicated 16-channel breast coil, and then compared the results with galactography. Histopathological diagnoses and follow-ups were used as reference standard. Lesion size estimated on MRI was compared with the size at histopathology. RESULTS: Sensitivity and specificity of MRI vs. galactography for detecting pathologic findings were 95.7 % vs. 85.7 % and 69.7 % vs. 33.3 %, respectively. For the supposed concrete pathology based on MRI findings, the specificity was 67.6 % and the sensitivity 77.3 % (PPV 60.7 %, NPV 82.1 %). Eight malignant lesions were detected (14.8 %). The estimated size at breast MRI showed excellent correlation with the size at histopathology (Pearson's correlation coefficient 0.95, p < 0.0001). CONCLUSIONS: MRI of the breast at 3.0 Tesla is an accurate imaging test and can replace galactography in the workup of nipple discharge in patients with inconspicuous mammography and ultrasound. KEY POINTS: • Breast MRI is an excellent diagnostic tool for patients with nipple discharge. • MRI of the breast reveals malignant lesions despite inconspicuous mammography and ultrasound. • MRI of the breast has greater sensitivity and specificity than galactography. • Excellent correlation of lesion size measured at MRI and histopathology was found.

Analytical and Clinical Validation of the Oncomine Dx Target Test to Assess HER2 Mutation Status in Tumor Tissue Samples From Patients With Non-Small Cell Lung Cancer Treated With Trastuzumab Deruxtecan in the DESTINY-Lung01 and DESTINY-Lung02 Studies.

CONTEXT.­: Trastuzumab deruxtecan (T-DXd), a human epidermal growth factor receptor 2 (HER2)-targeted therapy, has demonstrated durable anticancer activity in patients with advanced, metastatic HER2 (also known as ERBB2)-mutant (HER2m) non-small cell lung cancer (NSCLC) who have limited treatment options and poor prognosis. OBJECTIVE.­: To analytically validate and assess the clinical utility of the Oncomine Dx Target (ODxT) Test as a companion diagnostic to identify patients with HER2m NSCLC. DESIGN.­: Tumor samples from patients in DESTINY-Lung01 and DESTINY-Lung02 were retrospectively analyzed alongside commercially procured samples using the ODxT test and compared to the assays used for screening in these clinical trials. RESULTS.­: Positive percent agreement (PPA) and negative percent agreement (NPA) between the ODxT Test and TruSight Tumor 170 assay when testing DESTINY-Lung01 and commercially procured samples met prespecified thresholds (PPA and NPA ≥90%) for analytical accuracy (100% and 99.1%). The ODxT Test results were highly concordant with clinical trial assays (CTAs) used in DESTINY-Lung01 (PPA and NPA, 98.0% and 100%) and DESTINY-Lung02 (PPA and NPA, 96.7% and 100%) to identify activating HER2 mutations in tumor samples. Confirmed objective response rates were similar between patients with HER2m tumors identified by the ODxT Test and by CTAs in DESTINY-Lung01 (58.3% and 54.9%) and DESTINY-Lung02 (53.6% and 53.8%). Response duration was 12.0 and 9.3 months for patients identified by the ODxT Test and CTAs, respectively, in DESTINY-Lung01. CONCLUSIONS.­: The ODxT Test detected HER2 mutations in NSCLC with high analytical and clinical accuracy and identified HER2m populations with response rates similar to populations identified by CTAs, supporting clinical utility of the ODxT Test to inform treatment decisions for HER2m NSCLC.

Super-leadership and work enjoyment: direct and moderated influences.

Super-leadership is part of an approach called 'empowering leadership.' Within this approach, super-leadership is assumed to enable subordinates to lead themselves. The current study examined correlates of super-leadership. A questionnaire measuring two dimensions of super-leadership was used to analyze relationships between super-leadership and subordinates' work enjoyment, i.e., job satisfaction, subjective well-being, and emotional organizational commitment. In addition, moderating effects of the organizational context, i.e., organizational decentralization, on the relationships between super-leadership and work enjoyment were explored. 198 German employees from different occupations participated in the study. Latent moderator structural equation analysis revealed that the two factors of super-leadership, "coaching and communicative support" and "facilitation of personal autonomy and responsibility," had direct positive effects on subordinates' work enjoyment. Organizational decentralization moderated the effect of "coaching and communicative support" on work enjoyment but not the relations involving "facilitation of personal autonomy and responsibility." Conclusions for further research and practical applications were discussed.

Clinical Validation of Companion Diagnostics for the Selection of Patients with Non-Small Cell Lung Cancer Tumors Harboring Epidermal Growth Factor Receptor Exon 20 Insertion Mutations for Treatment with Amivantamab.

Amivantamab, an epidermal growth factor receptor (EGFR)-c-Met bispecific antibody, targets activating/resistance EGFR mutations and MET mutations/amplifications. In the ongoing CHRYSALIS study (ClinicalTrials.gov Identifier: NCT02609776), amivantamab demonstrated antitumor activity in patients with non-small cell lung cancer harboring EGFR exon 20 insertion mutations (ex20ins) that progressed on or after platinum-based chemotherapy, a population in which amivantamab use has been approved by the US Food and Drug Administration. This bridging study clinically validated two novel candidate companion diagnostics (CDx) for use in detecting EGFR ex20ins in plasma and tumor tissue, Guardant360 CDx and Oncomine Dx Target Test (ODxT), respectively. From the 81 patients in the CHRYSALIS efficacy population, 78 plasma and 51 tissue samples were tested. Guardant360 CDx identified 62 positive (16 negative), and ODxT identified 39 positive (3 negative), samples with EGFR ex20ins. Baseline demographic and clinical characteristics were similar between the CHRYSALIS-, Guardant360 CDx-, and ODxT-identified populations. Agreement with local PCR/next-generation sequencing tests used for enrollment into CHRYSALIS demonstrated high adjusted negative (99.6% and 99.9%) and positive (100% for both) predictive values with the Guardant360 CDx and ODxT tests, respectively. Overall response rates were comparable between the CHRYSALIS, Guardant360 CDx, and ODxT populations. Both the plasma- and tissue-based diagnostic tests provided accurate, comprehensive, and complementary approaches to identifying patients with EGFR ex20ins who could benefit from amivantamab therapy.

Optimized density-weighted imaging for dynamic contrast-enhanced 3D-MR mammography.

PURPOSE: To increase the spatial coverage and to reduce slice crosstalk combined with an optimal signal-to-noise ratio (SNR) in 3D dynamic contrast-enhanced (DCE) magnetic resonance (MR) mammography. MATERIALS AND METHODS: Asymmetric sampling schemes and a new reconstruction strategy based on virtual coils are presented for density-weighted (DW) 3D imaging. Additionally, for MR mammography an alternating DW (ADW) sampling along the k(y) direction shifts the undersampling artifacts out of the signal reception region. Virtual coils for effective DW (VIDED) imaging suppresses the aliasing in undersampled DW imaging. VIDED and ADW were compared to the conventional Cartesian imaging in phantom and in vivo MR mammography studies. RESULTS: The slice crosstalk was significantly reduced by VIDED and compared to Cartesian imaging the SNR increased by 16%. Additionally, VIDED and ADW provided a substantially increased field of view (FOV) in the slice direction and allowed the spatial resolution to be improved (up to 60% for ADW and 30% for VIDED) without lengthening the scan time. CONCLUSION: VIDED and ADW improve the image quality in 3D DCE MR mammography by enhancing the spatial resolution, reducing the slice crosstalk at nearly optimal SNR, and increasing the FOV in the slice direction. For VIDED no lengthening of the scan time or usage of multichannel receiver coils is necessary.

The Role of Motivational Regulation in Exam Preparation: Results From a Standardized Diary Study.

Previous studies have shown that the use of motivational regulation strategies has the potential to sustain invested effort and persistence in the learning process. Combining different methods (questionnaires and standardized diaries), the present study aimed to determine the role of motivational regulation in an exam preparation period. Motivational regulation is differentiated in a quantitative (extent of strategy use) and a qualitative (planning, implementing, monitoring, and correcting strategy use) aspect. One hundred and fifteen university students reported the quantity and quality of their motivational regulation strategy use in a pretest and kept a standardized learning diary focused on motivational difficulties and invested effort over a 14-day period just before an exam in their studies. Exam performance was assessed afterward. Results revealed positive effects of both aspects of motivational regulation on invested effort in exam preparation and exam performance. Moreover, a high quality of motivational regulation was associated with reduced negative effects of motivational difficulties on invested effort during studying-implying that motivational regulation can buffer against specific motivational problems occurring in the learning process.

Transcription and mRNA export machineries SAGA and TREX-2 maintain monoubiquitinated H2B balance required for DNA repair.

DNA repair is critical to maintaining genome integrity, and its dysfunction can cause accumulation of unresolved damage that leads to genomic instability. The Spt-Ada-Gcn5 acetyltransferase (SAGA) coactivator complex and the nuclear pore-associated transcription and export complex 2 (TREX-2) couple transcription with mRNA export. In this study, we identify a novel interplay between human TREX-2 and the deubiquitination module (DUBm) of SAGA required for genome stability. We find that the scaffold subunit of TREX-2, GANP, positively regulates DNA repair through homologous recombination (HR). In contrast, DUBm adaptor subunits ENY2 and ATXNL3 are required to limit unscheduled HR. These opposite roles are achieved through monoubiquitinated histone H2B (H2Bub1). Interestingly, the activity of the DUBm of SAGA on H2Bub1 is dependent on the integrity of the TREX-2 complex. Thus, we describe the existence of a functional interaction between human TREX-2 and SAGA DUBm that is key to maintaining the H2B/HB2ub1 balance needed for efficient repair and HR.

Genes and Pathways Regulated by Androgens in Human Neural Cells, Potential Candidates for the Male Excess in Autism Spectrum Disorder.

BACKGROUND: Prenatal exposure to androgens during brain development in male individuals may participate to increase their susceptibility to develop neurodevelopmental disorders such as autism spectrum disorder (ASD) and intellectual disability. However, little is known about the action of androgens in human neural cells. METHODS: We used human neural stem cells differentiated from embryonic stem cells to investigate targets of androgens. RESULTS: RNA sequencing revealed that treatment with dihydrotestosterone (DHT) leads to subtle but significant changes in the expression of about 200 genes, encoding proteins of extracellular matrix or involved in signal transduction of growth factors (e.g., insulin/insulin growth factor 1). We showed that the most differentially expressed genes (DEGs), RGCC, RNF144B, NRCAM, TRIM22, FAM107A, IGFBP5, and LAMA2, are reproducibly regulated by different androgens in different genetic backgrounds. We showed, by overexpressing the androgen receptor in neuroblastoma cells SH-SY5Y or knocking it down in human neural stem cells, that this regulation involves the androgen receptor. A chromatin immunoprecipitation combined with direct sequencing analysis identified androgen receptor-bound sequences in nearly half of the DHT-DEGs and in numerous other genes. DHT-DEGs appear enriched in genes involved in ASD (ASXL3, NLGN4X, etc.), associated with ASD (NRCAM), or differentially expressed in patients with ASD (FAM107A, IGFBP5). Androgens increase human neural stem cell proliferation and survival in nutrient-deprived culture conditions, with no detectable effect on regulation of neurite outgrowth. CONCLUSIONS: We characterized androgen action in neural progenitor cells, identifying DHT-DEGs that appear to be enriched in genes related to ASD. We also showed that androgens increase proliferation of neuronal precursors and protect them from death during their differentiation in nutrient-deprived conditions.

Comparison of ximelagatran with warfarin for the prevention of venous thromboembolism after total knee replacement.

BACKGROUND: In a previous study of the prevention of venous thromboembolism after total knee replacement, the efficacy of ximelagatran, an oral direct thrombin inhibitor that does not require monitoring of coagulation or dose adjustment, was found to be similar to that of warfarin at a dose of 24 mg of ximelagatran twice daily. The purpose of the present study was to determine whether a higher dose of ximelagatran is superior to warfarin. METHODS: This randomized, double-blind trial compared a regimen of 7 to 12 days of oral ximelagatran, at a dose of 24 or 36 mg twice daily, starting the morning after surgery, with warfarin therapy started the evening of the day of surgery. The composite end point of venous thromboembolism and death from all causes and the incidence of bleeding were the primary outcome measures. RESULTS: Among the 1851 patients in the efficacy analysis, oral ximelagatran at a dose of 36 mg twice daily was superior to warfarin with respect to the primary composite end point of venous thromboembolism and death from all causes (20.3 percent vs. 27.6 percent; P=0.003). There were no significant differences between these two groups with respect to major bleeding (incidence, 0.8 percent and 0.7 percent, respectively), perioperative indicators of bleeding, wound characteristics, or the composite secondary end point of proximal deep-vein thrombosis, pulmonary embolism, and death (2.7 percent vs. 4.1 percent; P=0.17). CONCLUSIONS: The efficacy of oral ximelagatran, administered starting the morning after total knee replacement, was superior to that of warfarin for prevention of venous thromboembolism. Rates of hemorrhagic complications with the two drugs were similar.

Evaluation of the nephrotoxicity of iodixanol in patients with predisposing factors to contrast medium induced nephropathy referred for contrast enhanced computed tomography.

To determine the risk of developing contrast induced nephropathy (CIN) in intermediate-risk patients receiving iodixanol, an iso-osmolar, dimeric non-ionic contrast agent, for CT in a clinical setting. Hundred consecutive patients referred for a contrast enhanced CT with a serum creatinine concentration>1.1mg/dl and/or a glomerular filtration rate (GFR)<90ml/min were included. Exclusion criteria were a serum creatinine>2.0mg/dl and a GFR<30ml/min or concurrent nephrotoxic agents. Between 60 and 140ml (mean 97+/-42ml) iodixanol (320mgI/ml) were administered at a flow of 2.5-3ml/s. Hydration with 500ml NaCl i.v. was performed before and after contrast injection. Follow-up was completed in 99 patients (age, 64+/-13 years, 68 men). CIN was defined as increase in serum creatinine concentration +0.5mg/dl or >25% above baseline within 72h after contrast administration. Serum creatinine concentration and GFR were 1.40+/-0.22, 1.29+/-0.29, and 1.26+/-0.29mg/dl and 52.2+/-13.9, 51.3+/-21.1, and 51.5+/-15.1ml/min on days 0, 3, and 7, respectively. Three out of 99 (3%) patients who received 90-110ml iodixanol revealed a CIN on day 3 without persistence on day 7. No specific therapy was needed. One out of 99 patients reported an exanthema on days 3 and 7. With the use of iodixanol in intermediate-risk patients, 3% of the patients develop CIN on day 3 without need for a specific therapy or persistence on day 7.

A prospective study comparing different iodine concentrations for triphasic multidetector row CT of the upper abdomen.

OBJECTIVES: To investigate the effect of different iodine concentrations at either constant injection or iodine administration rates but constant total iodine load on contrast enhancement of liver, pancreas and spleen by multidetector row CT. MATERIALS AND METHODS: One hundred and twenty consecutive patients (70+/-6 years) underwent triphasic liver CT at a four-channel multidetector-row CT using the non-ionic contrast medium iopromide. Patients were divided into six equal groups-I: 150 ml, 240 mg/ml at 4 ml/s; II: 120 ml, 300 mg/ml at 4 ml/s; III: 97.3 ml, 370 mg/ml at 4 ml/s; IV: 150 ml, 240 mg/ml at 5 ml/s; V: 120 ml, 300 mg/ml, 60 ml at 6 ml/s, 60 ml at 3 ml/s; VI: 97.3 ml, 370 mg/ml at 3.3 ml/s. ROIs were measured in the liver, the pancreas, and the spleen in unenhanced, arterial, portal venous, and equilibrium phase. RESULTS: At a constant injection rate of 4 ml/s, pancreatic enhancement over baseline only in the arterial phase was significantly higher at 370 mg/ml (58+/-15 HU versus 59+/-18 HU versus 74+/-20 HU for groups I-III, respectively (p<0.02)). Comparison of different iodine concentrations at constant iodine administration rate (groups II, IV and VI) and of all six protocols revealed no significant differences at either phase. CONCLUSIONS: At a constant iodine load and constant injection rates, the high-iodinated contrast agent iopromide at 370 mg/ml improves pancreatic enhancement in the arterial phase. At constant iodine load and constant iodine administration rates, there is no significant effect of different iodine concentrations.

Oral direct thrombin inhibitor ximelagatran compared with warfarin for the prevention of venous thromboembolism after total knee arthroplasty.

BACKGROUND: Warfarin, which requires coagulation monitoring, is associated with relatively high rates of thromboembolism despite providing adequate prophylaxis. This study compared an oral direct thrombin inhibitor, ximelagatran, with warfarin in order to evaluate the safety and efficacy of the medication for the prevention of venous thromboembolism in patients undergoing total knee arthroplasty. METHODS: Following surgery, patients were randomly assigned to fixed-dose oral ximelagatran (36 mg twice daily) or warfarin (target international normalized ratio, 2.5), both administered for seven to twelve days in a double-blind, double-dummy design. Warfarin was initiated on the evening of the day of surgery, and ximelagatran, on the morning after surgery. The primary efficacy end point was the incidence of asymptomatic deep-vein thrombosis determined by bilateral venography, objectively confirmed symptomatic deep-vein thrombosis or pulmonary embolism, and death from all causes during treatment. RESULTS: Adequate venograms or confirmed symptomatic events (efficacy population) were obtained for 1949 patients. Venous thromboembolism and death from all causes occurred in 22.5% (221) of 982 ximelagatran-treated patients and in 31.9% (308) of 967 warfarin-treated patients (p < 0.001). Proximal deep-vein thrombosis and pulmonary embolism were observed in 3.1% (thirty) and 0.2%, respectively, of the patients in the ximelagatran group and in 3.4% (thirty-three) and 0.4%, respectively, of the patients in the warfarin group. The six deaths from all causes included 0.3% (four) of the ximelagatran-treated patients and 0.2% (two) of the warfarin-treated patients. Major bleeding was noted in 1% (twelve) of the ximelagatran-treated patients and in 0.4% (five) of the warfarin-treated patients (p = 0.09). CONCLUSIONS: Oral ximelagatran (36 mg twice daily), administered without coagulation monitoring or dose adjustment and started the day after total knee arthroplasty, demonstrates superior efficacy compared with warfarin prophylaxis, with no wound complications and no significant difference with respect to bleeding events, although the rate of major bleeding events was greater with ximelagatran than with warfarin. LEVEL OF EVIDENCE: Therapeutic Level I.

Genomic landscapes of Chinese hamster ovary cell lines as revealed by the Cricetulus griseus draft genome.

Chinese hamster ovary (CHO) cells, first isolated in 1957, are the preferred production host for many therapeutic proteins. Although genetic heterogeneity among CHO cell lines has been well documented, a systematic, nucleotide-resolution characterization of their genotypic differences has been stymied by the lack of a unifying genomic resource for CHO cells. Here we report a 2.4-Gb draft genome sequence of a female Chinese hamster, Cricetulus griseus, harboring 24,044 genes. We also resequenced and analyzed the genomes of six CHO cell lines from the CHO-K1, DG44 and CHO-S lineages. This analysis identified hamster genes missing in different CHO cell lines, and detected >3.7 million single-nucleotide polymorphisms (SNPs), 551,240 indels and 7,063 copy number variations. Many mutations are located in genes with functions relevant to bioprocessing, such as apoptosis. The details of this genetic diversity highlight the value of the hamster genome as the reference upon which CHO cells can be studied and engineered for protein production.

Comparison of different iodine concentrations for multidetector row computed tomography angiography of segmental renal arteries.

The aims of this study were (1) to assess the diagnostic performance of multidetector row computed tomography angiography (CTA) on imaging of renal artery branches and (2) to investigate the effect of different iodine concentrations at constant total iodine load and either constant injection rates or constant iodine administration rates. A number of 120 consecutive patients (71+/-6 years of age) underwent CTA of renal arteries (collimation 4 x 1 mm) using the nonionic contrast medium iopromide, and were divided into six equal groups: 1: 150 ml, 240 mg/ml at 4 ml/s; 2: 120 ml, 300 mg/ml at 4 ml/s; 3: 97.3 ml, 370 mg/ml at 4 ml/s; 4: 150 ml, 240 mg/ml at 5 ml/s; 5: 120 ml, 300 mg/ml, 60 ml at 6 ml/s, 60 ml at 3 ml/s; 6: 97.3 ml, 370 mg/ml at 3.3 ml/s. The image quality of the main renal arteries (n=240) and their first-order to fourth-order branches was scored as 0 for no visualization, 1 for only visualization, and 2 for diagnostic. All main renal arteries were diagnostic. First-order branches had score 2 in 38/40, 40/40, 37/40, 38/40, 39/40, and 40/40 patients for groups 1-6, respectively (p=0.34). Second-order branches were imaged best in group 2 (p<0.002)). Third-order branches had score 2 in only 1/40, 5/40, 1/40, 2/40, 0/40, and 2/40 renal arteries. Fourth-order branches were not imaged diagnostically. At a constant total iodine load, the main renal arteries and their first-order branches achieved diagnostic image quality at all iodine concentrations in four-channel multidetector row CTA for the protocols tested. Second-order renal artery branches were imaged best at 120 ml contrast medium with an iodine concentration of 300 mg/ml at 4 ml/s.