Patient-specific cylinder templates for hybrid interstitial vaginal brachytherapy: Feasibility of automated 3-D design, 3D printing, and dosimetric outlook.

PURPOSE: Delivering highly conformal treatment plans for high-dose rate vaginal brachytherapy using commercially available applicators can be challenging. A partially automated workflow is presented for the in-house modeling and 3D printing of patient-specific cylindrical templates (PSCTs), which facilitate placement of flexible intracavitary and interstitial needles. To demonstrate feasibility, we compare PSCT treatment plans to retrospective interstitial brachytherapy plans delivered at our center. PSCTs derived from these plans were 3D printed to test the validity of the auto-design process. To facilitate clinical implementation, we validated the steam sterilization compatibility of PSCTs printed using polyetheretherketone (PEEK). METHODS AND MATERIALS: Plans for ten patients treated using a combination of vaginal cylinder and interstitial needles were compared to PSCT-based plans created for the same patient. DVH parameters for the HRCTV (V100, V150, V200, D90) and OARs (D2 cm3) were evaluated, as well as the number of needles used and the total interstitial length. Each planned PSCT was printed and compared to the intended needle geometry. 3D printed models were sterilization validated by an independent contractor for an autoclave protocol. RESULTS: PSCT plans demonstrated advantages over template based perineal BT in reducing the total interstitial needle length required while preserving or improving HRCTV and OAR dosimetry. All printed PSCTs matched planned geometry. CONCLUSIONS: PSCTs stand to be an alternative to current HDR-BT templates/applicators for patients with vaginal and locally recurrent endometrial cancers. Clinically equivalent or improved treatment plans can be created and devices to deliver these plans can be accurately printed and sterilized.

Evaluation of the ITV-margin and variables affecting bladder and mesorectal deformation during long course neoadjuvant radiotherapy for rectal cancer.

Internal target volume (ITV) margins were estimated by evaluating the movement of mesorectum and bladder during neoadjuvant long-course radiation therapy (RT) for rectal cancer. In this prospective study, 23 patients with rectal cancer had planning CT (pCT) and weekly cone beam CT (CBCT) in supine position during preoperative long-course RT. Mesorectal wall motion was analyzed based on the coordinates of the most anterior, posterior, left and right points on the pCT and CBCT. Overlap volume (OV) between the pCT bladder and CBCT mesorectum was generated. Variables that might affect relative bladder volume (ratio of CBCT to pCT bladder volumes), anterior mesorectal wall position, and OV were studied. ITV margins were also calculated. In females, smaller OV and less movement of the upper anterior mesorectal wall were identified, suggesting smaller ITV margins might be required compared to males. The relative bladder volume did not change significantly over time and was correlated with OV: the larger the relative bladder volume, the less the OV. ITV margin of 0.8 to 1.1 cm in right-left direction is satisfactory. Posteriorly, only 8 to 9 mm margin is required for upper and mid rectal regions but double of this is required for inferior third. Anteriorly, 1.3 cm margin is adequate for lower and mid rectal regions and 2.4 cm is required superiorly. An anisotropic ITV expansion of clinical target volume (CTV) for rectal cancer radiotherapy contouring provides a robust method to encompass the deformation of bladder and mesorectum. The ITV margin should take into account sex and distance from the anal verge.

A case of incidental STUMP discovery in a patient with concurrent prostatic adenocarcinoma.

Stromal tumours of the prostate are exceedingly rare, often presenting in patients in their fifth decade of life. They are classified as either stromal sarcomas, or stromal tumours of uncertain malignant potential (STUMP), the latter of which is known to have diverse clinical behaviour and thus surgical excision is often warranted. We present a case of a 71-year-old male, initially worked up by his family doctor due to mild obstructive voiding symptoms. Following a more thorough urologic workup, including a prostate biopsy, he was found to have a markedly elevated prostate specific antigen and positive cores on prostate biopsy demonstrating prostatic adenocarcinoma. The decision was made to treat with retropubic radical prostatectomy and bilateral pelvic lymph node dissection. Resulting pathology showed concurrent prostatic adenocarcinoma in addition to STUMP. The patient continues to be followed by oncology as well as a sarcoma specialist due to the unique nature of his case.

The addition of interstitial needles to intracavitary applicators in the treatment of locally advanced cervical cancer: Why is this important and how to implement in low- and middle-income countries?

PURPOSE: Cervical cancer is the leading cause of cancer mortality of women in low-/middle-income countries. Interstitial needles improve outcomes but require resources beyond those available in endemic regions. We conducted a retrospective review of the use of interstitial needles in locally advanced cervical cancer and simulated both 3D planning without needles and 2D planning to explore the benefit of interstitial needles. METHODS AND MATERIALS: 57 brachytherapy plans of 17 patients who had intracavitary tandem and ring plus interstitial brachytherapy were reviewed. Prescribed dose was 7 Gy × four fractions. 2D plans prescribed to point A were generated to represent a standard Manchester loading. Dosimetric outcomes to clinical target volume and organs at risk (OARs) were compared with those of 3D-based plans. RESULTS: High-risk clinical target volume coverage was excellent: 93.2% for 2D plans, 93.9% for 3D plans without needles, and 96.2% for 3D with needles. The mean dose to 90% of target was 8.5 Gy/fraction for 2D plans, 7.5 for 3D without needles, and 7.9 Gy/fraction for 3D with needles. However, the 2D plans delivered 12% above recommended dose constraints for OARs (except rectum). Dosimetric differences were found between 3D planning and 3D with needles for target coverage (p = 0.002). Dose to OARs was significantly lower when 3D plans with needles were compared with 2D plans. CONCLUSIONS: Interstitial needles provide an optimal therapeutic ratio for patients with high-volume disease or/and unfavorable topography. This justifies additional capital investment in resources for implementation to provide optimal treatment for locally advanced cervical cancer globally.

Association of human papilloma virus status and response to radiotherapy in vulvar squamous cell carcinoma.

INTRODUCTION: Vulvar squamous cell carcinoma develops through two separate pathways, associated with the presence or absence of high-risk human papilloma virus (HPV). The objective of this study was to evaluate treatment response and clinical outcomes in women with HPV-associated versus HPV-independent vulvar squamous cell carcinoma treated with primary radiation therapy, in order to determine the ability to use HPV status as a predictor of response to radiation therapy. METHODS: This was a retrospective cohort study combining data from British Columbia Cancer, Canada and Duke University, USA. Patients were included who had been treated with radiation therapy but excluded if they had received major surgical interventions. Immunohistochemistry for p16 (as a surrogate for high-risk HPV infection) and p53 was performed. We analyzed the univariable association between p16 status and clinico-pathological features and performed univariable survival analysis for p16. RESULTS: Forty-eight patients with vulvar squamous cell carcinoma treated with primary radiation therapy were identified: 26 p16 positive/HPV-associated patients and 22 p16 negative/HPV-independent patients. p16 positive vulvar squamous cell carcinoma demonstrated a significantly improved overall survival (HR 0.39, p=0.03) and progression-free survival (HR 0.35, p=0.02). In women treated with definitive radiation therapy, p16 positivity was associated with improved overall survival (HR 0.29, p<0.01) and progression-free survival (HR 0.21, p<0.01). Among patients who received sensitizing chemotherapy, a significant association was observed with p16 positive tumors and overall survival (HR 0.25, p=0.03) and progression-free survival (HR 0.09, p<0.01). CONCLUSION: This study suggests that HPV status in vulvar squamous cell carcinoma has both prognostic and predictive implications, with increased radiosensitivity demonstrated in HPV-associated vulvar squamous cell carcinoma. Implications may include radiation dose de-escalation for HPV-associated vulvar squamous cell carcinoma and increased surgical aggressiveness for HPV-independent vulvar squamous cell carcinoma.

Elucidation of Nuclear and Organellar Genomes of Gossypium hirsutum: Furthering Studies of Species Evolution and Applications for Crop Improvement.

Plant genomes are larger and more complex than other eukaryotic organisms, due to small and large duplication events, recombination and subsequent reorganization of the genetic material. Commercially important cotton is the result of a polyploidization event between Old and New World cottons that occurred over one million years ago. Allotetraploid cotton has properties that are dramatically different from its progenitors-most notably, the presence of long, spinnable fibers. Recently, the complete genome of a New World cotton ancestral species, Gossypium raimondii, was completed. Future genome sequencing efforts are focusing on an Old World progenitor, G. arboreum. This sequence information will enable us to gain insights into the evolution of the cotton genome that may be used to understand the evolution of other plant species. The chloroplast genomes of multiple cotton species and races have been determined. This information has also been used to gain insight into the evolutionary history of cotton. Analysis of the database of nuclear and organellar sequences will facilitate the identification of potential genes of interest and subsequent development of strategies for improving cotton.

Plasma cell densities and glomerular filtration rates predict renal allograft outcomes following acute rejection.

The contribution of T cells and graft-reactive antibodies to acute allograft rejection is widely accepted, but the role of graft-infiltrating B and plasma cells is controversial. We examined 56 consecutive human renal transplant biopsies classified by Banff schema into T-cell-mediated (N = 21), antibody-mediated (N = 18), and mixed (N = 17) acute rejection, using standard immunohistochemistry for CD3, CD20, CD138, and CD45. In a predominantly African-American population (75%), neither Banff classification nor C4d deposition predicted the return to dialysis. Immunohistochemical analysis revealed CD3(+) T cells as the dominant cell type, followed by CD20(+) B cells and CD138(+) plasma cells in all acute rejection types. Using univariate Cox Proportional Hazard analysis, plasma cell density significantly predicted graft failure while B-cell density trended toward significance. Surprisingly T-cell density did not predict graft failure. The estimated glomerular filtration rate (eGFR) at diagnosis of acute rejection also predicted graft failure, while baseline eGFR ≥6 months prior to biopsy did not. Using multivariate analysis, a model including eGFR at biopsy and plasma cell density was most predictive of graft loss. These observations suggest that plasma cells may be a critical mediator and/or an independently sensitive marker of steroid-resistant acute rejection.

Expression of MiTF may be helpful in differentiating cellular neurothekeoma from plexiform fibrohistiocytic tumor (histiocytoid predominant) in a partial biopsy specimen.

BACKGROUND: Overlapping histopathologic features of cellular neurothekeoma (CNT) and plexiform fibrohistiocytic tumor (PFHT), when both are predominantly composed of histiocytoid cells, make distinction between these entities challenging. Some have suggested that CNT and PFHT are related entities. No prior study has demonstrated a reliable immunohistochemical panel to differentiate these entities. METHODS: Skin biopsies diagnosed as CNT and PFHT, from 2004 to 2010 were retrieved with accompanying pathology reports. Each case was reviewed by at least 2 dermatopathologists and 2 soft tissue pathologists for confirmation of diagnosis. All cases were then evaluated for immunohistochemical expression of PAX2, NKIC3, CD10, and microphthalmia transcription factor (MiTF). RESULTS: Histopathologically, the histiocytoid areas of each tumor shared similar architecture, demonstrating nests and fascicles of histiocytoid to spindled cells, with some separation of nests by collagen bands. Both CNT and PFHT were uniformly positive for NKIC3 and CD10, and both were frequently PAX2 positive. MiTF was strongly and diffusely positive in CNT and was consistently negative in the PFHT. CONCLUSIONS: CNT and PFHT share many histopathologic features and immunohistochemical staining patterns. Of the stains we evaluated, we found that expression of MiTF may be a reliable marker for distinguishing CNT from histiocytoid-predominant PFHT, especially in instances where only a small part of the tumor is sampled for evaluation.

Histopathological predictors of renal function decrease after laparoscopic radical nephrectomy.

PURPOSE: Radical nephrectomy is inevitably associated with a variable renal function decrease. We assessed the association of histopathological parameters in nonneoplastic renal parenchyma with the renal function decrease after radical nephrectomy. MATERIALS AND METHODS: We evaluated 32 male and 17 female patients with a mean age of 55.9 years who underwent laparoscopic radical nephrectomy. Using the Cockcroft-Gault formula we calculated the estimated glomerular filtration rate preoperatively and at last followup at a mean of 19.7 months. The study end point was the percent change in the estimated glomerular filtration rate from baseline, defined as (absolute change/baseline) × 100. Three histological features in the nonneoplastic parenchyma were assessed by a renal pathologist, including global glomerulosclerosis, arteriosclerosis and interstitial fibrosis/tubular atrophy. For glomerulosclerosis assessment the percent of affected glomeruli was determined. Arteriosclerosis or the extent of arterial luminal occlusion was graded into 4 groups, including 1-0% to 5%, 2-6% to 25%, 3-26% to 50% and 4-greater than 50%. However, due to small patient numbers groups 1 and 2, and 3 and 4 were condensed, and AS was statistically evaluated as 0% to 25% or greater than 25%. Interstitial fibrosis/tubular atrophy was evaluated as absent/present. RESULTS: The mean estimated glomerular filtration rate decreased 31% from 122 to 85 ml/minute/1.73 m(2) after surgery (p < 0.0001). The percent change in the estimated glomerular filtration rate was associated with glomerulosclerosis extent (p = 0.034). For each 10% increase in glomerulosclerosis the estimated glomerular filtration rate decreased by 9% from baseline. The extent of arteriosclerosis or the presence of interstitial fibrosis/tubular atrophy did not correlate with the estimated glomerular filtration rate decrease. CONCLUSIONS: Glomerulosclerosis severity in nonneoplastic parenchyma can predict the rate of renal function decrease after radical nephrectomy. This histopathological parameter should be assessed in all tumor nephrectomy specimens, given that preserving renal function is important for quality of life and clinical outcome in patients with renal cancer.

Bruno negatively regulates germ cell-less expression in a BRE-independent manner.

Mechanisms of post-transcriptional control are essential during Drosophila oogenesis and embryogenesis to sequester gene products in discrete regions and ultimately achieve embryonic asymmetry. Maternal germ cell-less (gcl) mRNA accumulates in the pole plasm of the embryo before Gcl protein is detectable. gcl mRNA, but not Gcl protein, can also be detected in somatic regions of the embryo, suggesting that gcl RNA is subject to translational control. We find that Gcl is expressed during oogenesis, and that it is regulated by the translational repressor Bruno (Bru). Increased levels of Gcl are observed in the oocyte when Bru level is reduced, and overexpression of Bru reduces Gcl expression. Consistently, reduction of the maternal dosage of Bruno leads to ectopic Gcl expression in the embryo, which, in turn, represses anterior hückebein (hkb) expression. Bru binds directly to the gcl 3'UTR in vitro, but, surprisingly, this binding is independent of a BRE (Bruno response element)-like motif. This motif is also not required for in vivo repression of Gcl expression during oogenesis or early embryogenesis. Bru binds the gcl 3'UTR via its C-terminal domain, which includes RNA recognition motif 3 (RRM3), with little or no contribution from the remainder of the protein. We conclude that repression by Bruno during oogenesis is required to restrict Gcl expression in the early embryo and that Bru represses gcl expression in a manner that involves RRM3 and a sequence unrelated to the BRE.

Breaking the A chain: regulating mRNAs in development through CCR4 deadenylase.

Post-transcriptional mechanisms of gene regulation have long been implicated in specifying embryonic pattern in many organisms. Experiments in Caenorhabditis elegans, Drosophila, and Xenopus have recently converged, pointing to the CCR4 deadenylase complex as a key effector that modulates the expression of proteins from specific germline mRNAs.