Hnf4a Is Required for the Development of Cdh6-Expressing Progenitors into Proximal Tubules in the Mouse Kidney.

BACKGROUND: Hepatocyte NF 4α (Hnf4a) is a major regulator of renal proximal tubule (PT) development. In humans, a mutation in HNF4A impairs PT functions and is associated with Fanconi renotubular syndrome (FRTS). In mice, mosaic deletion of Hnf4a in the developing kidney reduces the population of PT cells, leading to FRTS-like symptoms. The molecular mechanisms underlying the role of Hnf4a in PT development remain unclear. METHODS: The gene deletion tool Osr2Cre removed Hnf4a in developing nephrons in mice, generating a novel model for FRTS. Immunofluorescence analysis characterized the mutant phenotype, and lineage analysis tested whether Cadherin-6 (Cdh6)-expressing cells are PT progenitors. Genome-wide mapping of Hnf4a binding sites and differential gene analysis of Hnf4a mutant kidneys identified direct target genes of Hnf4a. RESULTS: Deletion of Hnf4a with Osr2Cre led to the complete loss of mature PT cells, lethal to the Hnf4a mutant mice. Cdh6high, lotus tetragonolobus lectin-low (LTLlow) cells serve as PT progenitors and demonstrate higher proliferation than Cdh6low, LTLhigh differentiated PT cells. Additionally, Hnf4a is required for PT progenitors to differentiate into mature PT cells. Genomic analyses revealed that Hnf4a directly regulates the expression of genes involved in transmembrane transport and metabolism. CONCLUSIONS: Hnf4a promotes the differentiation of PT progenitors into mature PT cells by regulating the expression of genes associated with reabsorption, the major function of PT cells.

Notch is required for the formation of all nephron segments and primes nephron progenitors for differentiation.

Notch signaling plays important roles during mammalian nephrogenesis. To investigate whether Notch regulates nephron segmentation, we performed Notch loss-of-function and gain-of-function studies in developing nephrons in mice. Contrary to the previous notion that Notch signaling promotes the formation of proximal tubules and represses the formation of distal tubules in the mammalian nephron, we show that inhibition of Notch blocks the formation of all nephron segments and that constitutive activation of Notch in developing nephrons does not promote or repress the formation of a specific segment. Cells lacking Notch fail to form the S-shaped body and show reduced expression of Lhx1 and Hnf1b Consistent with this, we find that constitutive activation of Notch in mesenchymal nephron progenitors causes ectopic expression of Lhx1 and Hnf1b and that these cells eventually form a heterogeneous population that includes proximal tubules and other types of cells. Our data suggest that Notch signaling is required for the formation of all nephron segments and that it primes nephron progenitors for differentiation rather than directing their cell fates into a specific nephron segment.

Early clinical outcomes of helical tomotherapy/intensity-modulated proton therapy combination in nasopharynx cancer.

This study aimed to evaluate the feasibility of combining helical tomotherapy (HT) and intensity-modulated proton therapy (IMPT) in treating patients with nasopharynx cancer (NPC). From January 2016 to March 2018, 98 patients received definitive radiation therapy (RT) with concurrent chemotherapy (CCRT). Using simultaneous integrated boost and adaptive re-plan, 3 different dose levels were prescribed: 68.4 Gy in 30 parts to gross tumor volume (GTV), 60 Gy in 30 parts to high-risk clinical target volume (CTV), and 36 Gy in 18 parts to low-risk CTV. In all patients, the initial 18 fractions were delivered by HT, and, after rival plan evaluation on the adaptive re-plan, the later 12 fractions were delivered either by HT in 63 patients (64.3%, HT only) or IMPT in 35 patients (35.7%, HT/IMPT combination), respectively. Propensity-score matching was conducted to control differences in patient characteristics. In all patients, grade ≥ 2 mucositis (69.8% vs 45.7%, P = .019) and grade ≥ 2 analgesic usage (54% vs 37.1%, P = .110) were found to be less frequent in HT/IMPT group. In matched patients, grade ≥ 2 mucositis were still less frequent numerically in HT/IMPT group (62.9% vs 45.7%, P = .150). In univariate analysis, stage IV disease and larger GTV volume were associated with increased grade ≥ 2 mucositis. There was no significant factor in multivariate analysis. With the median 14 month follow-up, locoregional and distant failures occurred in 9 (9.2%) and 12 (12.2%) patients without difference by RT modality. In conclusion, comparable early oncologic outcomes with more favorable acute toxicity profiles were achievable by HT/IMPT combination in treating NPC patients.

Notch signaling promotes nephrogenesis by downregulating Six2.

During nephrogenesis, multipotent mesenchymal nephron progenitors develop into distinct epithelial segments. Each nephron segment has distinct cell types and physiological function. In the current model of kidney development, Notch signaling promotes the formation of proximal tubules and represses the formation of distal tubules. Here, we present a novel role of Notch in nephrogenesis. We show in mice that differentiation of nephron progenitors requires downregulation of Six2, a transcription factor required for progenitor maintenance, and that Notch signaling is necessary and sufficient for Six2 downregulation. Furthermore, we find that nephron progenitors lacking Notch signaling fail to differentiate into any nephron segments, not just proximal tubules. Our results demonstrate how cell fates of progenitors are regulated by a transcription factor governing progenitor status and by a differentiation signal in nephrogenesis.

Clinical implementation of Dosimetry Check™ for TomoTherapy® delivery quality assurance.

PURPOSE: The delivery quality assurance (DQA) of intensity-modulated radiotherapy (IMRT) plans is a prerequisite for ensuring patient treatments. This work investigated the clinical usefulness of a new DQA system, Dosimetry Check™(DC), on TomoTherapy® -based helical IMRT plans. METHODS: The DQA was performed for 15 different TomoTherapy® -based clinical treatment plans. In Tomotherapy® machines, the couch position was set to a height of 400 mm and the treatment plans were delivered using QA-Treatment mode. For each treatment plan, the plan data and measured beam fluence were transferred to a DC-installed computer. Then, DC reconstructed the three-dimensional (3D) dose distribution to the CT images of the patient. The reconstructed dose distribution was compared with that of the original plan in terms of absolute dose, two-dimensional (2D) planes and 3D volume. The DQA results were compared with those performed by a conventional method using the cheese phantom with ion chamber and radiochromic film. RESULTS: For 14 out of the 15 treatment plans, the absolute dose difference between the measurement and calculation was less than 3% and the gamma pass rate with the 3%/3 mm gamma evaluation criteria was greater than 95% for both DQA methods. The P-value calculated using Wilcoxon signed-rank test was 0.256, which implies no statistically significance in determining the absolute dose difference between the two methods. For one treatment plan generated using the 5.0 cm field width, the absolute dose difference was greater than 3% and the gamma pass rate was less than 95% with DC, while the DQA result with the cheese phantom method passed our TomoTherapy® DQA tolerance. CONCLUSION: We have clinically implemented DC for the DQA of TomoTherapy® -based helical IMRT treatment plans. DC carried out the accurate DQA results as performed with the conventional cheese phantom method. This new DQA system provided more information in verifying the dose delivery to patients, while simplifying the DQA process.

Single cell dissection of early kidney development: multilineage priming.

We used a single cell RNA-seq strategy to create an atlas of gene expression patterns in the developing kidney. At several stages of kidney development, histologically uniform populations of cells give rise to multiple distinct lineages. We performed single cell RNA-seq analysis of total mouse kidneys at E11.5 and E12.5, as well as the renal vesicles at P4. We define an early stage of progenitor cell induction driven primarily by gene repression. Surprising stochastic expression of marker genes associated with differentiated cell types was observed in E11.5 progenitors. We provide a global view of the polarized gene expression already present in the renal vesicle, the first epithelial precursor of the nephron. We show that Hox gene read-through transcripts can be spliced to produce intergenic homeobox swaps. We also identify a surprising number of genes with partially degraded noncoding RNA. Perhaps most interesting, at early developmental times single cells often expressed genes related to several developmental pathways. This provides powerful evidence that initial organogenesis involves a process of multilineage priming. This is followed by a combination of gene repression, which turns off the genes associated with most possible lineages, and the activation of increasing numbers of genes driving the chosen developmental direction.

Characterization and evaluation of an integrated quality monitoring system for online quality assurance of external beam radiation therapy.

PURPOSE: The aim of this work was to comprehensively evaluate a new large field ion chamber transmission detector, Integral Quality Monitor (IQM), for online external photon beam verification and quality assurance. The device is designed to be mounted on the linac accessory tray to measure and verify photon energy, field shape, gantry position, and fluence before and during patient treatment. METHODS: Our institution evaluated the newly developed ion chamber's effect on photon beam fluence, response to dose, detection of photon fluence modification, and the accuracy of the integrated barometer, thermometer, and inclinometer. The detection of photon fluence modifications was performed by measuring 6 MV with fields of 10 cm × 10 cm and 1 cm × 1 cm "correct" beam, and then altering the beam modifiers to simulate minor and major delivery deviations. The type and magnitude of the deviations selected for evaluation were based on the specifications for photon output and MLC position reported in AAPM Task Group Report 142. Additionally, the change in ion chamber signal caused by a simulated IMRT delivery error is evaluated. RESULTS: The device attenuated 6 MV, 10 MV, and 15 MV photon beams by 5.43 ± 0.02%, 4.60 ± 0.02%, and 4.21 ± 0.03%, respectively. Photon beam profiles were altered with the IQM by < 1.5% in the nonpenumbra regions of the beams. The photon beam profile for a 1 cm × 1 cm2 fields were unchanged by the presence of the device. The large area ion chamber measurements were reproducible on the same day with a 0.14% standard deviation and stable over 4 weeks with a 0.47% SD. The ion chamber's dose-response was linear (R2 = 0.99999). The integrated thermometer agreed to a calibrated thermometer to within 1.0 ± 0.7°C. The integrated barometer agreed to a mercury barometer to within 2.3 ± 0.4 mmHg. The integrated inclinometer gantry angle measurement agreed with the spirit level at 0 and 180 degrees within 0.03 ± 0.01 degrees and 0.27 ± 0.03 at 90 and 270 degrees. For the collimator angle measurement, the IQM inclinometer agreed with a plum-bob within 0.3 ± 0.2 degrees. The simulated IMRT error increased the ion chamber signal by a factor of 11-238 times the baseline measurement for each segment. CONCLUSIONS: The device signal was dependent on variations in MU delivered, field position, single MLC leaf position, and nominal photon energy for both the 1 cm × 1 cm and 10 cm × 10 cm fields. This detector has demonstrated utility repeated photon beam measurement, including in IMRT and small field applications.

A bioinformatics method identifies prominent off-targeted transcripts in RNAi screens.

Because off-target effects hamper interpretation and validation of RNAi screen data, we developed a bioinformatics method, genome-wide enrichment of seed sequence matches (GESS), to identify candidate off-targeted transcripts in primary screening data. GESS analysis revealed a prominent off-targeted transcript in several screens, including MAD2 (MAD2L1) in a screen for genes required for the spindle assembly checkpoint. GESS analysis results can enhance the validation rate in RNAi screens.

Effects of bisphenol A and triclocarban on brain-specific expression of aromatase in early zebrafish embryos.

Estrogen regulates numerous developmental and physiological processes. Most effects are mediated by estrogen receptors (ERs), which function as ligand-regulated transcription factors. Estrogen also regulates the activity of GPR30, a membrane-associated G protein-coupled receptor. Many different types of environmental contaminants can activate ERs; some can bind GPR30 as well. There is growing concern that exposure to some of these compounds, termed xenoestrogens, is interfering with the behavior and reproductive potential of numerous wildlife species, as well as affecting human health. Here, we investigated how two common, environmentally pervasive chemicals affect the in vivo expression of a known estrogen target gene in the brain of developing zebrafish embryos, aromatase AroB, which converts androgens to estrogens. We confirm that, like estrogen, the well-studied xenoestrogen bisphenol A (BPA, a plastics monomer), induces strong brain-specific overexpression of aromatase. Experiments using ER- and GPR30-selective modulators argue that this induction is largely through nuclear ERs. BPA induces dramatic overexpression of AroB RNA in the same subregions of the developing brain as estrogen. The antibacterial triclocarban (TCC) by itself stimulates AroB expression only slightly, but TCC strongly enhances the overexpression of AroB that is induced by exogenous estrogen. Thus, both BPA and TCC have the potential to elevate levels of aromatase and, thereby, levels of endogenous estrogens in the developing brain. In contrast to estrogen, BPA-induced AroB overexpression was suppressed by TCC. These results indicate that exposures to combinations of certain hormonally active pollutants can have outcomes that are not easily predicted from their individual effects.

Monte Carlo investigation of collapsed versus rotated IMRT plan verification.

IMRT QA requires, among other tests, a time-consuming process of measuring the absorbed dose, at least to a point, in a high-dose, low-dose-gradient region. Some clinics use a technique of measuring this dose with all beams delivered at a single gantry angle (collapsed delivery), as opposed to the beams delivered at the planned gantry angle (rotated delivery). We examined, established, and optimized Monte Carlo simulations of the dosimetry for IMRT verification of treatment plans for these two different delivery modes (collapsed versus rotated). The results of the simulations were compared to the treatment planning system dose calculations for the two delivery modes, as well as to measurements taken. This was done in order to investigate the validity of the use of a collapsed delivery technique for IMRT QA. The BEAMnrc, DOSXYZnrc, and egs_chamber codes were utilized for the Monte Carlo simulations along with the MMCTP system. A number of different plan complexity metrics were also used in the analysis of the dose distributions in a bid to qualify why verification in a collapsed delivery may or may not be optimal for IMRT QA. Following the Alfonso et al. formalism, the kfclin,frefQclin,Q correction factor was calculated to correct the deviation of small fields from the reference conditions used for beam calibration. We report on the results obtained for a cohort of 20 patients. The plan complexity was investigated for each plan using the complexity metrics of homogeneity index, conformity index, modulation complexity score, and the fraction of beams from a particular plan that intersect the chamber when performing the QA. Rotated QA gives more consistent results than the collapsed QA technique. The kfclin,frefQclin,Qfactor deviates less from 1 for rotated QA than for collapsed QA. If the homogeneity index is less than 0.05 then the kfclin,frefQclin,Q factor does not deviate from unity by more than 1%. A value this low for the homogeneity index can only be obtained with the rotated QA technique.

Hedgehog signaling is required for the maintenance of mesenchymal nephron progenitors.

Background: Mesenchymal nephron progenitors (mNPs) give rise to all nephron tubules in the mammalian kidney. Since premature depletion of these cells leads to low nephron numbers, high blood pressure, and various renal diseases, it is critical to understand how mNPs are maintained. While Fgf, Bmp, and Wnt signaling pathways are known to be required for the maintenance of these cells, it is unclear if any other signaling pathways also play roles. Methods: To test the potential role of Hedgehog signaling in mNPs, we conditionally deleted Shh from the collecting duct and Smo from the nephron lineage. To identify the genes regulated by Hedgehog signaling in mNPs, we performed RNA-seq analysis from mNPs with different Smo doses. To test if the upregulation of Notch signaling mimics loss of Hedgehog signaling, we performed Jag1 gain-of-function study in mNPs. Results: We found that loss of either Shh or Smo resulted in premature depletion of mNPs. Our transcriptional profiling data from Smo loss- and gain-of-function mutant mNPs suggested that Hedgehog signaling inhibited the activation of Notch signaling and upregulated the expression of Fox transcription factors such as Foxc1 and Foxp4. Consistent with these observations, we found that ectopic expression of Jag1 caused the premature depletion of mNPs as seen in the Smo mutant kidney. We also found that Foxc1 was capable of binding to mitotic condensed chromatin, a feature of a mitotic bookmarking factor. Conclusions: Our study demonstrates a previously unappreciated role of Hedgehog signaling in preventing premature depletion of mNPs by repressing Notch signaling and likely by activating the expression of Fox factors.

Single-cell sequencing dissects the transcriptional identity of activated fibroblasts and identifies novel persistent distal tubular injury patterns in kidney fibrosis.

Examining kidney fibrosis is crucial for mechanistic understanding and developing targeted strategies against chronic kidney disease (CKD). Persistent fibroblast activation and tubular epithelial cell (TEC) injury are key CKD contributors. However, cellular and transcriptional landscapes of CKD and specific activated kidney fibroblast clusters remain elusive. Here, we analyzed single cell transcriptomic profiles of two clinically relevant kidney fibrosis models which induced robust kidney parenchymal remodeling. We dissected the molecular and cellular landscapes of kidney stroma and newly identified three distinctive fibroblast clusters with "secretory", "contractile" and "vascular" transcriptional enrichments. Also, both injuries generated failed repair TECs (frTECs) characterized by decline of mature epithelial markers and elevation of stromal and injury markers. Notably, frTECs shared transcriptional identity with distal nephron segments of the embryonic kidney. Moreover, we identified that both models exhibited robust and previously unrecognized distal spatial pattern of TEC injury, outlined by persistent elevation of renal TEC injury markers including Krt8 and Vcam1, while the surviving proximal tubules (PTs) showed restored transcriptional signature. We also found that long-term kidney injuries activated a prominent nephrogenic signature, including Sox4 and Hox gene elevation, which prevailed in the distal tubular segments. Our findings might advance understanding of and targeted intervention in fibrotic kidney disease.

Dose homogeneity specification for reference dosimetry of nonstandard fields.

PURPOSE: To investigate the sensitivity of the plan-class specific correction factor to dose distributions in composite nonstandard field dosimetry. METHODS: A cylindrical water-filled PMMA phantom was constructed at the center of which reference absorbed dose could be measured. Ten different TomoTherapy(®)-based IMRT fields were created on the CT images of the phantom. The dose distribution for each IMRT field was estimated at the position of a radiation detector or ionization chamber. The dose in each IMRT field normalized to that in a reference 10 × 10 cm(2) field was measured using a PTW micro liquid ion chamber. Based on the new dosimetry formalism, a plan-class specific correction factor k(Q(pcsr),Q) (f(pcsr),f(ref)) for each field was measured for two Farmer-type chambers, Exradin A12 and NE2571, as well as for a smaller Exradin A1SL chamber. The dependence of the measured correction factor on parameters characterizing dose distribution was analyzed. RESULTS: Uncertainty on the plan-class specific correction factor measurement was in the range of 0.3%-0.5% and 0.3%-0.8% for the Farmer-type chambers and the Exradin A1SL, respectively. When the heterogeneity of the central region of the target volume was less than 5%, the correction factor did not differ from unity by more than 0.7% for the three air-filled ionization chambers. For more heterogeneous dose deliveries, the correction factor differed from unity by up to 2.4% for the Farmer-type chambers. For the Exradin A1SL, the correction factor was closer to unity due to the reduced effect of dose gradients, while it was highly variable in different IMRT fields because of a more significant impact of positioning uncertainties on the response of this chamber. CONCLUSIONS: The authors have shown that a plan-class specific correction factor can be specified as a function of plan evaluation parameters especially for Farmer-type chambers. This work provides a recipe based on quantifying dose distribution to accurately select air-filled ionization chamber correction factors for nonstandard fields.

Toxicity of Proton Therapy versus Photon Therapy on Salvage Re-Irradiation for Non-Small Cell Lung Cancer.

This study evaluated the toxicity associated with radiation techniques on curative re-irradiation (re-RT) in patients with thoracic recurrence of non-small cell lung cancer (NSCLC). From 2011 to 2019, we retrospectively reviewed the data of 63 patients with salvage re-RT for in-field or marginal recurrence of NSCLC at two independent institutions. Re-RT techniques using X-ray beams and proton beam therapy (PBT) were also included. Re-RT had a 2-year overall survival (OS) and local progression-free survival of 48.0% and 52.0%, respectively. Fifteen patients experienced grade 3 or higher toxicity after re-RT. The complication rates were 18.2% (4/22) and 26.8% (11/41) in PBT patients and X-ray patients, respectively. Airway or esophageal fistulas occurred in seven patients (11.1%). Fistulas or severe airway obstruction occurred in patients with tumors adjacent to the proximal bronchial tree and esophagus, who underwent hypofractionated radiotherapy (RT) or concurrent chemotherapy, and with a higher dose exposure to the esophagus. In conclusion, salvage re-RT was feasible even in patients with local recurrence within the previous RT field. PBT showed similar survival outcomes and toxicity to those of other techniques. However, thoracic re-RT should be performed carefully considering tumor location and RT regimens such as the fraction size and concurrent chemotherapy.

Phosphorylation of Cdc20 is required for its inhibition by the spindle checkpoint.

The spindle checkpoint delays anaphase until all chromosomes are properly attached to spindle microtubules. When the spindle checkpoint is activated at unattached kinetochores, the checkpoint proteins BubR1, Bub3 and Mad2 bind and inhibit Cdc20, an activator of the anaphase-promoting complex (APC). Here, we show that Xenopus laevis Cdc20 is phosphorylated at Ser 50, Thr 64, Thr 68 and Thr 79 during mitosis and that mitogen-activated protein kinase (MAPK) contributes to the phosphorylation at Thr 64 or Thr 68. Cdc20 mutants that are phosphorylation-deficient are able to activate the APC in X. laevis egg extracts. However, Cdc20 mutants in which any of the four phosphorylation sites were altered to Ala or Val failed to respond to the spindle checkpoint signal, owing to their reduced affinity for the spindle checkpoint proteins. This study demonstrates that the spindle checkpoint stops anaphase by inhibiting fully-phosphorylated Cdc20. Our results also have implications for the spindle checkpoint silencing mechanism.

Monitor unit prediction model for wobbling proton therapy with ridge filters.

PURPOSE: We introduced an output factor (cGy/MU) prediction model for wobbling proton beams over the full range of proton energy, scatterer thickness, and the width of spread-out Bragg peak (SOBP). MATERIALS AND METHODS: From December 2015 to August 2020, 1990 wobbling proton fields were used to treat patients, where 1714 fields had a diameter smaller than 11 cm and 276 had a diameter between 11 and 16 cm, which were designated as small and middle wobbling radius cases, respectively. The output factor is defined as the ratio of proton absorbed dose at mid-depth of SOBP to monitor unit (MU). It depends dominantly on proton energy, scatterer thickness, and the width of SOBP. We established the prediction model using the polynomial fitting function and determined its coefficients for the small and middle wobbling radius cases. We evaluated the accuracy of our prediction model by calculating the difference between predicted and measured output factors. RESULTS: For the small wobbling radius cases, the mean value of the output factor difference was 0.22% with a standard deviation of 1.3%. For the middle wobbling radius cases, the mean value was 0.20% and with a standard deviation of 0.79%. The large deviation was especially observed for wobbling proton beams having small field size and small width of SOBP. CONCLUSIONS: We made a prediction model of output factor for wobbling proton beams, thereby determining MU of each beam. This included the dependency of the output factor on the proton energy between 70 and 230 MeV, scatterer thickness, and the width of SOBP. For 93.6% of the small and 95.5% of the middle wobbling radius cases, the deviation between predicted and measured output factor was below 3%. The cases with deviations of predicted and measured output factor above 3% had small field size and small width of SOBP. The accuracy of our prediction model would be improved by adopting the field size effect and measuring more cases of small field size and small SOBP width in the future.

Investigation of three radiation detectors for accurate measurement of absorbed dose in nonstandard fields.

PURPOSE: To establish accurate experimental dosimetry techniques for reference dose measurements in nonstandard composite fields. METHODS: A cylindrical PMMA phantom filled with water was constructed, at the center of which reference absorbed dose to water for a head and neck IMRT delivery was measured. Based on the proposed new formalism for reference dosimetry of nonstandard fields [Alfonso et al., Med. Phys. 35, 5179-5186 (2008)], a candidate plan-class specific reference (pcsr) field for a typical head and neck IMRT delivery was created on the CT images of the phantom. The absorbed dose to water in the pcsr field normalized to that in a reference 10 x 10 cm2 field was measured using three radiation detectors: Gafchromic EBT films, a diamond detector, and a guarded liquid-filled ionization chamber developed in-house (GLIC-03). Pcsr correction factors [formula in text] were determined for five different types of air-filled ionization chambers (Exradin A12, NE-2571, Exradin A1SL, Exradin A14, and PinPoint 31006) in a fully rotated delivery and in a delivery with the same MLC settings and weights but from a single gantry angle (a collapsed delivery). RESULTS: The combined standard uncertainty in measuring the correction factor [formula in text] using the three dosimetry techniques was 0.3%. For all the air-filled ionization chambers and the pcsr field tested, the correction factor was not different from unity by more than +/- 0.8%. For the fully rotated delivery, the correction factors were in a narrow range of 0.9955-0.9986, while in the collapsed delivery, they were in a slightly broader range of 0.9922-1.0048. In the collapsed delivery, the Farmer-type chambers (Exradin A12 and NE2571) had very similar correction factors (0.9922 and 0.9931, respectively), whereas the correction factors for the smaller chambers showed more distinct chamber-type dependence. CONCLUSIONS: The authors have established three experimental dosimetry techniques that allow reference measurements of nonstandard field correction factors [formula in text] for air-filled ionization chambers at the 0.3% 1sigma uncertainty level. These techniques can be used to determine criteria for the selection of plan-class specific reference fields and ultimately improve clinical reference dosimetry of nonstandard fields.

ß-catenin regulates the formation of multiple nephron segments in the mouse kidney.

The nephron is composed of distinct segments that perform unique physiological functions. Little is known about how multipotent nephron progenitor cells differentiate into different nephron segments. It is well known that ß-catenin signaling regulates the maintenance and commitment of mesenchymal nephron progenitors during kidney development. However, it is not fully understood how it regulates nephron segmentation after nephron progenitors undergo mesenchymal-to-epithelial transition. To address this, we performed ß-catenin loss-of-function and gain-of-function studies in epithelial nephron progenitors in the mouse kidney. Consistent with a previous report, the formation of the renal corpuscle was defective in the absence of ß-catenin. Interestingly, we found that epithelial nephron progenitors lacking ß-catenin were able to form presumptive proximal tubules but that they failed to further develop into differentiated proximal tubules, suggesting that ß-catenin signaling plays a critical role in proximal tubule development. We also found that epithelial nephron progenitors lacking ß-catenin failed to form the distal tubules. Expression of a stable form of ß-catenin in epithelial nephron progenitors blocked the proper formation of all nephron segments, suggesting tight regulation of ß-catenin signaling during nephron segmentation. This work shows that ß-catenin regulates the formation of multiple nephron segments along the proximo-distal axis of the mammalian nephron.

Dummy Run of Quality Assurance Program before Prospective Study of Hippocampus-Sparing Whole-Brain Radiotherapy and Simultaneous Integrated Boost for Multiple Brain Metastases from Non-small Cell Lung Cancer: Korean Radiation Oncology Group (KROG) 17-06 Study.

PURPOSE: Lung Cancer Subcommittee of Korean Radiation Oncology Group (KROG) has recently launched a prospective clinical trial (KROG 17-06) of hippocampus-sparing whole brain radiotherapy (HS-WBRT) with simultaneous integrated boost (SIB) in treating multiple brain metastases from non-small cell lung cancer. In order to improve trial quality, dummy run studies among the participating institutions were designed. This work reported the results of two-step dummy run procedures of the KROG 17-06 study. MATERIALS AND METHODS: Two steps tested hippocampus contouring variability and radiation therapy planning compliance. In the first step, the variation of the hippocampus delineation was investigated for two representative cases using the Dice similarity coefficients. In the second step, the participating institutions were requested to generate a HS-WBRT with SIB treatment plan for another representative case. The compliance of the treatment plans to the planning protocol was evaluated. RESULTS: In the first step, the median Dice similarity coefficients of the hippocampus contours for two other dummy run cases changed from 0.669 (range, 0.073 to 0.712) to 0.690 (range, 0.522 to 0.750) and from 0.291 (range, 0.219 to 0.522) to 0.412 (range, 0.264 to 0.598) after providing the hippocampus contouring feedback. In the second step, with providing additional plan priority and extended dose constraints to the target volumes and normal structures, we observed the improved compliance of the treatment plans to the planning protocol. CONCLUSION: The dummy run studies demonstrated the notable inter-institutional variability in delineating the hippocampus and treatment plan generation, which could be decreased through feedback from the trial center.

Hnf4a deletion in the mouse kidney phenocopies Fanconi renotubular syndrome.

Different nephron tubule segments perform distinct physiological functions, collectively acting as a blood filtration unit. Dysfunction of the proximal tubule segment can lead to Fanconi renotubular syndrome (FRTS), with major symptoms such as excess excretion of water, glucose, and phosphate in the urine. It has been shown that a mutation in HNF4A is associated with FRTS in humans and that Hnf4a is expressed specifically in proximal tubules in adult rat nephrons. However, little is known about the role of Hnf4a in nephrogenesis. Here, we found that Hnf4a is expressed in both presumptive and differentiated proximal tubules in the developing mouse kidney. We show that Hnf4a is required for the formation of differentiated proximal tubules but is dispensable for the formation of presumptive proximal tubules. Furthermore, we show that loss of Hnf4a decreased the expression of proximal tubule-specific genes. Adult Hnf4a mutant mice presented with FRTS-like symptoms, including polyuria, polydipsia, glycosuria, and phosphaturia. Analysis of the adult Hnf4a mutant kidney also showed proximal tubule dysgenesis and nephrocalcinosis. Our results demonstrate the critical role of Hnf4a in proximal tubule development and provide mechanistic insight into the etiology of FRTS.