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Photoinduced vitamin D formation occurs 10–15-fold faster in phospholipid bilayers (PLB) than in isotropic solution. It has been hypothesized that amphipatic interactions of the PLB with the rotationally flexible previtamin D (Pre) stabilize its helical conformers, enhancing thermal intramolecular [1,7]-hydrogen transfer, forming vitamin D. To test this hypothesis, we carried out molecular dynamics (MD) simulations of Pre in a PLB composed of dipalmitoylphosphatidylcholine (DPPC). We designed a classical force field capable of accurately describing the equilibrium composition of Pre conformers. Using adaptive biasing force MD simulations, we determined the free energy of Pre conformers in isotropic environments (hexane and gas-phase) and in the anisotropic environment of a DPPC PLB. We find a total increase of 25.5% of the population of both helical conformers (+20.5% g+Zg+ and +5% g–Zg−) in DPPC compared to hexane. In view of ab initio simulations, showing that hydrogen transfer occurs in both helical conformers, our study strongly suggests the validity of the initial hypothesis. Regarding the amphipatic interactions of Pre with the PLB, we find that, similar to cholesterol (Chol) and 7-dehydrocholesterol (7-DHC), Pre entertains hydrogen bonds mainly to the carbonyl groups of DPPC and, to a lesser extent, with phosphate oxygen atoms and rarely to water molecules at the interface. We further report order parameters of the Pre/DPPC system, which are slightly smaller than those for Chol/DPPC and 7-DHC/DPPC, but larger than for pure DPPC. This indicates a loss in membrane viscosity upon photochemical ring-opening of 7-DHC to form Pre.

Mucinous ovarian carcinoma is an uncommon malignancy characterized by resistance to chemotherapy and poor survival in the metastatic setting. HER2 amplification is a frequent late event in carcinogenesis, yet the incidence of HER2-low in mucinous ovarian carcinoma is unknown. Further, the optimal method for determining overexpression in these tumors is not established. We sought to assess the ASCO/CAP and ToGA trial scoring methods for HER2 IHC with correlation to FISH, p53, and mismatch repair protein status and to determine the incidence of HER2-low in mucinous ovarian carcinoma. A total of 29 tumors from 23 patients were included. Immunohistochemistry for HER2, p53, MLH1, PMS2, MSH2, and MSH6 was performed. Scoring was performed according to the ASCO/CAP and ToGA trial criteria. HER2 FISH was performed and scored according to the ASCO/CAP criteria. The proportion of HER2-low, defined as 1+ or 2+ staining with negative FISH, was determined. Using ASCO/CAP, 26% demonstrated 3+ while 35% demonstrated 2+ staining. Using ToGA, 30% demonstrated 3+ while 57% demonstrated 2+ staining. By FISH, 26% were positive for HER2 amplification. Both systems captured all FISH-positive cases; the use of ASCO/CAP resulted in fewer equivocal and false-positive cases. Among HER2-negative cases, 88% were HER2-low. Aberrant p53 expression was detected in 55% of cases; mismatch repair deficiency was not identified in any cases. ASCO/CAP guidelines are accurate and resource-effective in determining HER2 overexpression in mucinous ovarian carcinoma. HER2-low is common in these tumors; further studies to determine the role of HER2-targeted therapy including antibody-drug conjugates are indicated.

US law continues its historical trend of growing more permissive towards actors who engage in violent action in purported self-defense. We draw on some informal game theory to show why this is strategically irrational and suggest rolling back self-defense doctrines like stand your ground to earlier historical precedents like duty to retreat.

Cytogenetics has long represented a critical component in the clinical evaluation of hematologic malignancies. Chromosome banding studies provide a simultaneous snapshot of genome-wide copy number and structural variation, which have been shown to drive tumorigenesis, define diseases, and guide treatment. Technological innovations in sequencing have ushered in our present-day clinical genomics era. With recent publications highlighting novel sequencing technologies as alternatives to conventional cytogenetic approaches, we, an international consortium of laboratory geneticists, pathologists, and oncologists, describe herein the advantages and limitations of both conventional chromosome banding and novel sequencing technologies and share our considerations on crucial next steps to implement these novel technologies in the global clinical setting for a more accurate cytogenetic evaluation, which may provide improved diagnosis and treatment management. Considering the clinical, logistic, technical, and financial implications, we provide points to consider for the global evolution of cytogenetic testing.

Byline: Adam C. Smith A new paternalism is on the rise. Using the insights of behavioral economics, agencies like the Environmental Protection Agency and the Consumer Financial Protection Bureau have [...]

Gastric-type endocervical adenocarcinoma is an uncommon aggressive type of endocervical adenocarcinoma that is not associated with human papillomavirus (HPV). At present, this tumor is classified under the spectrum of mucinous carcinoma of the uterine cervix. The clinical stage of gastric-type endocervical adenocarcinoma at the time of diagnosis is usually more advanced compared to the HPV-associated endocervical adenocarcinoma. Widespread dissemination to unusual sites, such as omentum, peritoneum, and distant organs, can be present. Owing to its rare incidence, diagnostic dilemmas, and aggressive behavior, clinical management can be challenging. In this study, we aimed to elucidate the molecular characteristics of these tumors by using next-generation sequencing (NGS) to assess 161 unique cancer-driver genes for single-nucleotide and copy-number variations, gene fusions, and insertions/deletions within gastric-type endocervical adenocarcinoma tumors. In total, 92 variants were detected across the 14 samples tested (7 variants on average per tumor). TP53was the most recurrently mutated gene followed by MSH6, CDKN2A/B, POLE, SLX4, ARID1A, STK11, BRCA2,and MSH2. Abnormal p53 expression was observed in nine cases by immunohistochemistry, of which TP53variants were present in four cases. MDM2gene amplification in 12q15 (69202190-69233452) locus was seen in two cases that express normal p53 levels by immunohistochemistry. Four cases had STK11null (frameshift/nonsense) variants, three of which were previously reported in Peutz–Jeghers syndrome. Overall, genes that are implicated in DNA damage, repair, cell cycle, Fanconi anemia pathway, and the PI3K-AKT signaling pathways were found to be mutated. Of note, genes known to have acquired and/or inherited variants in endometrial tumors were enriched within our cohort. In conclusion, our study shows the genetic heterogeneity of gastric-type endocervical adenocarcinoma with some potentially actionable molecular alterations, which highlights the importance of further molecular characterization for better identification of this rare entity, and hence better clinical management.

Aqueous methylglyoxal chemistry has often been implicated as an important source of oligomers in atmospheric aerosol. Here we report on chemical analysis of brown carbon aerosol particles collected from cloud cycling/photolysis chamber experiments, where gaseous methylglyoxal and methylamine interacted with glycine, ammonium, or methylammonium sulfate seed particles. Eighteen N-containing oligomers were identified in the particulate phase by liquid chromatography/diode array detection/electrospray ionization high-resolution quadrupole time-of-flight mass spectrometry. Chemical formulas were determined and, for 6 major oligomer products, MS2fragmentation spectra were used to propose tentative structures and mechanisms. Electronic absorption spectra were calculated for six tentative product structures by an ab initio second order algebraic-diagrammatic-construction/density functional theory approach. For five structures, matching calculated and measured absorption spectra suggest that they are dominant light-absorbing species at their chromatographic retention times. Detected oligomers incorporated methylglyoxal and amines, as expected, but also pyruvic acid, hydroxyacetone, and significant quantities of acetaldehyde. The finding that ∼80% (by mass) of detected oligomers contained acetaldehyde, a methylglyoxal photolysis product, suggests that daytime methylglyoxal oligomer formation is dominated by radical addition mechanisms involving CH3CO*. These mechanisms are evidently responsible for enhanced browning observed during photolytic cloud events.

Cytoplasmic FMRP interacting protein 1 (CYFIP1) is a candidate gene for intellectual disability (ID), autism, schizophrenia and epilepsy. It is a member of a family of proteins that is highly conserved during evolution, sharing high homology with its Drosophila homolog, dCYFIP. CYFIP1 interacts with the Fragile X mental retardation protein (FMRP, encoded by the FMR1 gene), whose absence causes Fragile X syndrome, and with the translation initiation factor eIF4E. It is a member of the WAVE regulatory complex (WRC), thus representing a link between translational regulation and the actin cytoskeleton. Here, we present data showing a correlation between mRNA levels of CYFIP1 and other members of the WRC. This suggests a tight regulation of the levels of the WRC members, not only by post-translational mechanisms, as previously hypothesized. Moreover, we studied the impact of loss of function of both CYFIP1 and FMRP on neuronal growth and differentiation in two animal models – fly and mouse. We show that these two proteins antagonize each other's function not only during neuromuscular junction growth in the fly but also during new neuronal differentiation in the olfactory bulb of adult mice. Mechanistically, FMRP and CYFIP1 modulate mTor signaling in an antagonistic manner, likely via independent pathways, supporting the results obtained in mouse as well as in fly at the morphological level. Collectively, our results illustrate a new model to explain the cellular roles of FMRP and CYFIP1 and the molecular significance of their interaction.

Quantification of regional or national variation in population trends is an integral component of assessing species conservation status, and ideally uses spatial and temporal replicate surveys across the breeding or wintering ranges. However, populations of boreal‐breeding birds are often monitored using site‐specific trends in the number of individuals counted migrating through more populated regions en route to or from their breeding grounds. Combining data across sites to quantify regional variation in a population trend has not occurred, and would rely on the success of model selection to choose the model that best describes the underlying pattern of regional population change. Using simulated daily counts of migrating birds with known regional rate(s) of population trend, we tested the utility of Akaike's Information Criterion (AIC) for detecting regional variation in population trend by comparing a series of hierarchical models that estimated nationally or regionally varying trends. When trends varied regionally, AIC successfully ranked the model that best described the regional pattern of population trend as the top model 100% of the time using 40‐year time series, and with as few as 3 monitoring sites in each region. When trends did not vary among regions, the more highly parameterized (incorrect) model with regionally varying trends was ranked as the top model ≤20% of the time, regardless of the length of time series or number of sites in a region. In this case, the incorrect regional model resulted in less precise trends but also comparable or higher power and comparable or lower probability of drawing false inference from the data than did the correct model. Compared with sampling 10 sites/region over a 20‐year time period, sampling as few as 3 sites/region over a 40‐year time period can result in a lower probability of detecting a false trend. Promoting long‐term monitoring at fewer sites can also minimize the financial and logistical resources required to maintain migration monitoring programs, and allow more recent population change to be interpreted within the context of long‐term population fluctuations. © 2015 The Wildlife Society.

As a sustainability advocate, I was inspired by Wayne Curtis's article ('Houses of the Future,' December Atlantic) describing the various homebuilding techniques New Orleans is applying. The energy-efficient, small footprint [...]

For the logic generations of the 15 nm node and beyond, the printing of pitches at 64nm and below are needed. For EUV lithography to replace ArF-based multi-exposure techniques, it is required to print these patterns in a single exposure process. The k1factor is roughly 0.6 for 64nm pitch at an NA of 0.25, and k1 0.52 for 56nm pitch. These k1numbers are of the same order at which model based OPC was introduced in KrF and ArF lithography a decade or so earlier. While we have done earlier work that used model-based OPC for the 22nm node test devices using EUV,1we used a simple threshold model without further resist model calibration. For 64 nm pitch at an NA of 0.25, the OPC becomes more important, and at 56nm pitch it becomes critical. For 15 nm node lithography, we resort to a full resist model calibration using tools that were adapted from conventional optical lithography. We use a straight shrink 22 nm test layout to assess post-OPC printability of a metal layer at pitches at 64 nm and 56 nm, and we use this information to correct test layouts.

Autophagy mediates the degradation of cytoplasmic components in eukaryotic cells and plays a key role in immunity. The mechanism of autophagosome formation is not clear. Here we examined two potential membrane sources for antibacterial autophagy: the ER and mitochondria. DFCP1, a marker of specialized ER domains known as ‘omegasomes,’ associated with Salmonella-containing autophagosomes via its PtdIns(3)P and ER-binding domains, while a mitochondrial marker (cytochrome b5-GFP) did not. Rab1 also localized to autophagosomes, and its activity was required for autophagosome formation, clearance of protein aggregates and peroxisomes, and autophagy of Salmonella. Overexpression of Rab1 enhanced antibacterial autophagy. The role of Rab1 in antibacterial autophagy was independent of its role in ER-to-Golgi transport. Our data suggest that antibacterial autophagy occurs at omegasomes and reveal that the Rab1 GTPase plays a crucial role in mammalian autophagy.

Several letters to the editor are presented in response to articles in previous issues including the article "Shots in the Dark" by Shannon Brownlee and Jeanne Lenzer in the November 2009 issue, a response by Brownlee and Lenzer to a letter to the editor about their article "Shots in the Dark" in the November 2009 issue, and the article "Did Christianity Cause the Crash?" by Hanna Rosin in the December 2009 issue.

Over a 10year period blood samples were collected from 57 individuals with growth restriction and RSSlike features. Our goal was to identify epigenetic abnormalities in this cohort, including uniparental disomy of chromosome 7 UPD7, methylation changes at chromosome11p15, as well as new epigenomic alterations. We evaluated the methylation status of 7 imprinting control regions on chromosomes 7, 11, 14, and 15. UPD7 and chromosome 7 structural abnormalities had been previously identified in five patients. Epigenetic alterations on chromosome 11p15 were identified in 11 patients. Of interest, in 3 of these 11 patients, the epigenetic alterations were limited to the H19 promoter and the distal region of its associated imprinting center, ICR1. In addition, in one patient, we detected methylation changes consistent with maternal UPD at all tested imprinted regions. This patient series suggests that epimutations on chromosome 11p15 can be most efficiently detected in RSS patients by screening for DNA methylation defects at the H19 promoter or the distal region of ICR. © 2009 WileyLiss, Inc.

The Canadian Wildlife Service (CWS) requires reliable estimates of the harvest of migratory game birds, including waterfowl, to effectively manage populations of these hunted species. The National Harvest Survey is an annual survey of hunters who purchase Canada's mandatory migratory game bird hunting permit, integrating information from a survey of hunting activity with information from a separate survey of species composition in the harvest. We used these survey data to estimate the number of birds harvested for each species and hunting activity metrics (e.g., number of active hunters, days spent hunting). The analytical methods used to generate these estimates have not changed since the survey was first designed in the early 1970s. We describe a new hierarchical Bayesian integrated model, which replaces the series of ratio estimators that comprised the old model. We are using this new model to generate estimates for migratory bird harvests as of the 2019–2020 hunting season, and to generate updated estimates for all earlier years. The hierarchical Bayesian model uses over‐dispersed Poisson distributions to model mean hunter activity and harvest (zero‐inflated Poisson and zero‐truncated Poisson, respectively). It also includes multinomial distributions to model some key components (e.g., variation in harvest across periods of the hunting season, the species composition of the harvest within each of those periods, the age and sex composition in the harvests of a given species). We estimated the parameters of the Poisson and the multinomial distributions for each year as random effects using first‐difference time‐series. This time‐series component allows the model to share information across years and reduces the sensitivity of the estimates to annual sampling noise. The new model estimates are generally very similar to those from the old model, particularly for the species that occur most commonly in the harvest, so the results do not suggest any major changes to harvest management decisions and regulations. Estimates for all species from the new model are more precise and less susceptible to annual sampling error, particularly for species that occur less commonly in the harvest (e.g., sea ducks, other species of conservation concern). This new model, with its hierarchical Bayesian framework, will also facilitate future improvements and elaborations, allowing the incorporation of prior information from the rich literature and knowledge in game bird management and biology. This integrated hierarchical model provides improved estimates of the hunting activity and harvest of migratory game birds in Canada and new estimates for age‐ and sex‐specific harvest. The Bayesian framework and the open‐source code that we provide will facilitate ongoing improvements and elaborations of the model.

22q11 Deletion syndrome (22q11DS) is the most common microdeletion syndrome in humans, occurring with an incidence of 1 in 4,000. In most cases the submicroscopic deletion spans 3 Mb, but there are a number of other overlapping and non‐overlapping deletions that generate a similar phenotype. The majority of the 22q11.2 microdeletions can be ascertained using a standard fluorescence in situ hybridization (FISH) assay probing for TUPLE1 or N25 on 22q11.2. However, this test fails to detect deletions that are either proximal or distal to the FISH probes, and does not provide any information about the length of the deletion. In order to increase the detection rate of 22q11.2 deletion and to better characterize the size and position of such deletions we undertook a study of 22q11.2 cases using multiplex ligation dependent probe amplification (MLPA). We used MLPA to estimate the size of the 22q11.2 deletions in 51 patients positive for TUPLE1 or N25 (FISH) testing, and to investigate 12 patients with clinical features suggestive of 22q11DS and negative FISH results. MLPA analysis confirmed a microdeletion in all 51 FISH‐positive samples as well as microduplications in three samples. Further, it allowed us to delineate deletions not previously detected using standard clinical FISH probes in 2 of 12 subjects with clinical features suggestive of 22q11DS. We conclude that MLPA is a cost‐effective and accurate diagnostic tool for 22q11DS with a higher sensitivity than FISH alone. Additional advantages of MLPA testing in our study included determination of deletion length and detection of 22q11.2 duplications. © 2007 Wiley‐Liss, Inc.

Beckwith–Wiedemann syndrome (BWS) is an overgrowth syndrome characterized by macrosomia, macroglossia, omphalocele, hemihyperplasia, and increased tumor risk. BWS can be associated with genetic and/or epigenetic alterations that modify imprinted gene expression on chromosome 11p15.5. Somatic mosaicism for paternal uniparental disomy (UPD) of chromosome 11p15, found in 20% of BWS patients, is associated with specific features of BWS including hemihyperplasia, Wilms tumor, and hepatoblastoma. The highly variable phenotypic spectrum of BWS associated with UPD may well reflect the level of UPD 11 cells in specific organs and tissues such that very high levels of UPD might produce a more severe phenotypic expression of BWS. In this regard we report on two patients with severe presentations of BWS and extremely high levels of UPD in DNA from lymphocytes. Clinically, both patients demonstrated extreme macroglossia, persistent hypoglycemia, cardiomyopathy, hemihyperplasia, renal abnormalities, abdominal organomegaly, hepatoblastoma and died in the first 6 months of life. These two patients support the hypothesis that high levels of UPD define high expressivity in BWS. © 2007 Wiley‐Liss, Inc.

Genomic imprinting refers to parent-of-origin–specific gene expression. Human chromosome band 11p15.5 houses a large cluster of genes that are imprinted. Dysregulation of this gene cluster is associated with the overgrowth and tumor predisposition syndrome, Beckwith-Wiedemann syndrome. Several genes in this imprinted cluster encode proteins involved in growth regulation, e.g.the paternally expressed IGF2and the maternally expressed cell-cycle regulator cyclin dependent kinase inhibitor, CDKN1C. Disruption of imprinted gene expression can result from genetic or epigenetic alterations. Genetic alterations such as duplication, deletion, translocation, inversion, and mutation in imprinted regions have been shown to cause disease. In addition, epimutations that are extrinsic to the primary DNA sequence have also been shown to cause disease. These epimutations usually involve gain or loss of methylation at regulatory differentially methylated regions. Recently, several human diseases in addition to Beckwith-Wiedemann syndrome have been reported to have molecular alterations at chromosome 11p15.5. These include isolated hemihyperplasia, Russell-Silver syndrome, and transient neonatal diabetes mellitus. These molecular alterations and their phenotypic effects on growth are discussed.

Isolated hemihyperplasia (IH) refers to a distinct diagnosis involving asymmetric overgrowth of single or multiple organs or regions of the body and can result from various genomic changes including molecular alterations of 11p15; these are paternal uniparental disomy (UPD), and alterations of methylation at two imprinting centers at 11p15: IC1 (H19) and IC2 (KCNQ1OT1). As little information is available on the molecular basis of tumor development in IH, or on the frequency of tumors in children with different molecular subtypes of IH, molecular testing was undertaken on 51 patients with IH and revealed: 8 (16%) with UPD, 3 (6%) with hypomethylation at KCNQ1OT1, and 0 with hypermethylation at H19. Of the 8 patients with UPD, 4 had tumors (3 hepatoblastomas, 1 Wilms tumor); 0/3 patients with hypomethylation at KCNQ1OT1 had a tumor; of the remaining 40 with no molecular alterations, 6 had tumors (3 Wilms tumors, 2 neuroblastomas, 1 adrenocortical adenoma). The 50% tumor frequency in patients with IH and UPD was statistically significantly higher than the 15% tumor frequency in those with IH and no molecular alteration detected (Fisher's exact test P = 0.047, OR 5.67). This is the first demonstration that UPD at 11p15 in patients with IH confers a higher tumor risk than in patients with IH without this molecular change. Of note, two of the eight patients with UPD and IH were conceived using assisted reproductive technologies (ART), thus raising the question whether ART might impact the rate of somatic recombination during embryonic development. © 2006 Wiley‐Liss, Inc.

The clinical importance of overgrowth syndromes in the pediatric patient population has been increasingly recognized during the past decade, but clinical overlap among overgrowth syndromes often makes diagnostic categorization difficult. Advances in the molecular delineation of overgrowth syndromes in recent years have furthered our knowledge of the phenotypic spectrum of this group of conditions. This review focuses on developments in our understanding of the molecular mechanisms and phenotype-genotype correlations in the two most common overgrowth syndromes, Beckwith-Wiedemann syndrome and Sotos syndrome. The implications of these findings with respect to clinical diagnosis, medical management, and genetic counseling are discussed.

Beckwith–Wiedemann syndrome (BWS) is a clinically heterogeneous overgrowth syndrome associated with an increased risk for embryonal tumor development. BWS provides an ideal model system to study epigenetic mechanisms. This condition is caused by a variety of genetic or epigenetic alterations within two domains of imprinted growth regulatory genes on human chromosome 11p15. Molecular studies of BWS have provided important data with respect to epigenotype/genotype–phenotype correlations; for example, alterations of Domain 1 are associated with the highest risk for tumor development, specifically Wilms' tumor. Further, the elucidation of the molecular basis for monozygotic twinning in BWS defined a critical period for imprint maintenance during pre‐implantation embryonic development. In the future, such molecular studies in BWS will permit enhanced medical management and targeted genetic counseling. © 2005 Wiley‐Liss, Inc.

Rhabdomyosarcoma (RMS) is a soft tissue tumor of childhood frequently diagnosed between the first and fifth year of life. Children with the Beckwith-Wiedemann syndrome (BWS), a congenital overgrowth syndrome characterized by exomphalos, macroglossia, and macrosomia, have an increased risk of developing childhood tumors including Wilms tumor, hepatoblastoma, neuroblastoma, and RMS. Although an association between RMS and the BWS is well accepted, only four cases have been reported to date, and of these, three were reported as embryonal RMS. Based on these data, an association between BWS and embryonal RMS has been proposed. We report three additional cases of BWS with RMS and review the clinical data for each patient as well as the pathology of their tumors. All three cases of BWS had histology consistent with alveolar RMS and were diagnosed at 6 weeks and 5 and 13 years of age. In two of these BWS cases, constitutional defects of 11p15 imprinting were demonstrated. Furthermore, cytogenetic analysis of the tumors did not detect the t(2;13) or t(1;13) translocations that generate the PAX3- or PAX7-FKHR fusion proteins common to alveolar RMS. These observations suggest that the development of alveolar RMS tumors in BWS may occur without the chromosomal rearrangement producing the PAX-FKHR fusion protein. In summary, we present three new cases of RMS demonstrating a new association between BWS and an uncommon subtype of alveolar RMS. The absence of the translocations commonly associated with alveolar rhabdomyosarcoma suggests a common 11p15 pathway for alveolar RMS and BWS.

Introduction

Introduction

The photosynthesis of vitamin D3in mammalian skin results from UV-B irradiation of provitamin D3(7-dehydrocholesterol, DHC) at ca. 290 nm. Upon return to the ground state, the hexatriene product, previtamin D3, undergoes a conformational equilibration between helical gZg and more planar tZg and tZt forms. The helical gZg forms provide a pathway for the formation of vitamin D3via a [1,7]-sigmatropic hydrogen shift. Steady state photolysis and UV transient absorption spectroscopy are combined to explore the conformational relaxation of previtamin D3formed from DHC in isotropic solution and confined to lipid bilayers chosen to model the biological cell membrane. The results are compared with measurements for two analogues: previtamin D2formed from ergosterol (provitamin D2) and previtamin D3acetate formed from DHC acetate. The resulting spectral dynamics are interpreted in the context of simulations of optical excitation energy and oscillator strength as a function of conformation. In solution, the relaxation dynamics and steady state product distributions of the three compounds are nearly identical, favoring tZg forms. When confined to lipid bilayers, the heterogeneity and packing forces alter the conformational distributions and enhance the population of a gZg conformer capable of vitamin D formation.

Presents several letters to the editor. Review of the book "Kayaks You Can Build," by Ted Moores and Greg Rössel; View that unwavering trust, a keen sense of fun, a compromising nature, and an adventurous spirit are key factors to a successful boat partnership; Discussion on new boat technology.

Toze: Roche: Consultancy, Honoraria, Research Funding. Gerrie:F Hoffmann-La Roche: Other.

The article focuses on the announcement by U.S. Environmental Protection Agency's Administrator Gina McCarthy of the anti-coal-burning Clean Power Plan of U.S. President Barack Obama's administration. It discusses the support of the plan by the environmental community, and the benefit of the plan to Baptists and bootlegger, the benefit of the plan to dirty coal producers.

Important advances in the understanding of CLL pathogenesis include the discovery that NOTCH1 mutations are present in ∼28% of pts harboring +12. There is a need for improved understanding of the clinical outcomes of CLL patients (pts) with +12 on a population-level, as this subgroup is rapidly becoming the focus of biologic studies evaluating pathogenesis of disease and clinical trials investigating novel targeted therapies. In the province of BC, population 4.5 million, CLL pts receive uniform evaluation and therapy based on centrally derived protocols with FISH testing implemented since 2004. We sought to characterize the clinical outcomes of +12 in this large unselected population-based cohort of CLL pts.Clinical and laboratory data on all pts referred for CLL FISH testing at 1 of 3 BC cytogenetic labs from 2004–2011 were entered into the BC Provincial CLL Database and included in this analysis. Pts without a confirmed diagnostic date were excluded. Baseline features of pts with and without +12 were compared using Fisher's exact test for categorical and Wilcoxon rank sum test for continuous (cnts) variables. Primary and secondary endpoints were OS and TFS (defined as time from diagnosis [dx] to first therapy). Percent of abnormal (%abn) nuclei harboring +12 was evaluated for association with OS/TFS. Cox proportional hazard (PH) models were constructed to determine predictors of OS/TFS for the +12 cohort, including age at dx, sex, Rai stage (0, 1–2, 3–4), WBC at dx, CD38 positivity and concomitant 17p-, 11q- or deletion 13q (13q-). Cox PH models were also constructed to determine effect of +12 on TFS/OS for the entire cohort.As of Dec. 2011, 882 pts had CLL FISH testing in BC of which 164 (19%) had +12 on their 1st FISH test: 8 (5%) with concomitant 17p-; 14 (9%) with 11q-; 142 (86%) without either 17p- or 11q-, of which 43 (30%) had 13q-; 16/124 tested (13%) had an IGH translocation [t(IGH)]. Of the 164 +12 pts, median age at dx was 60 yrs (range 35–93), 70% were male, 10% had Rai stage 3–4. At median follow-up of 4.5 yrs (range 0–19), 95 pts (59%) received treatment, 31 (19%) died. For the +12 cohort, median OS was 14.7 yrs (95% CI 9.8–19.0) and median TFS 3.7 yrs (95% CI 2.7–5.4).Of the 658 non +12 CLL pts (N12CPs), prevalence of recurrent cytogenetic abnormalities (RCA) were: 17p-, 10%; 11q-, 11%; 13q-, 60%; t(IGH) 7%. Significant differences between +12 and N12CPs included more CD38+ pts (66% vs 28%, P<0.001), higher t(IGH) incidence (13% vs 7%, P=0.04) and fewer 17p- (5% vs 10%, P=0.03) or 13q- (26% vs 60%, P<0.001) abn among +12 pts. When pts were grouped by hierarchical FISH abn, +12 pts retained an intermediate OS (median 15.9 yrs) and TFS (median 4.2 yrs) when compared to other RCAs (Fig 1A). Multivariate analysis (MVA) for the whole cohort (n=822) demonstrated no significant effect of +12 on OS (HR 0.72, 95% CI 0.36–1.43, P=.35) or TFS (HR 0.86, 95% CI 0.69–1.36, P=.86) after adjustment for covariates.For the +12 cohort (n=162), univariate analysis demonstrated shorter OS associated with age (P=.001), Rai stage (P=.01) and 17p- (P=.07). A longer OS was associated with presence of 13q- (median OS 11.6 vs 18.7 yrs, P=.04), Fig 1B. Shorter TFS was associated with Rai stage (P<.001), WBC at dx (P=.01) and 17p- (P=.04). %abn nuclei harboring +12 was not predictive of OS (P=.33) or TFS (P=.25) as a cnts variable; however those with <20% vs ≥20% abn had a significant improvement in OS (P=.02). MVA for the +12 cohort demonstrated Rai stage (HR 3.26, 95% CI 1.23– 8.63, P=.02) and 11q- (HR 9.07, 95% CI 1.44–57.02, P=.02) as independent risk factors for OS, while 13q- did not retain its protective effect (P=.98). For TFS, MVA found Rai stage (HR 2.92, 95% CI 1.78–4.78, P<.001) and 17p- (HR 5.44, 95% CI 1.52–19.43, P=.01) as negative predictors while 13q- (HR 2.01, 95% CI 1.08–3.75, P=.03) again had a positive effect.We report the largest, population-based cohort of CLL pts with FISH testing and confirm that +12 occurs in 19% of CLL pts and in the absence of 17p- or 11q-, confers an intermediate prognosis. The presence of 13q- had a protective effect on TFS and a trend towards improved OS, thus improving the prognosis of a subset of +12 pts. This finding is consistent with recent observations that NOTCH1 mutations and 13q- are mutually exclusive in +12 pts and may explain the clinical heterogeneity seen in this subgroup. Further research into these distinct subsets of +12 pts is warranted.No relevant conflicts of interest to declare.

CLL has been considered a genetically stable disease; however, recent evidence suggests that clonal evolution (CE) occurs at a higher rate than previously known. The clinical significance of CE in terms of CLL progression, requirement for and response to therapy (tx), overall (OS) and treatment-free survival (TFS) is not clear. The relationship between CE and tx and to the pathophysiology of CLL is also currently unknown. There is a need for improved understanding of CE in CLL and its possible impact on prognosis, tx, and outcomes. We performed a population-based analysis of all patients (pts) referred for CLL fluorescence in situ hybridization (FISH) testing in BC, Canada who had 32 FISH studies done to determine the incidence of CE and its association with clinical outcomes.FISH testing has been standard of care for CLL pts in BC since 2004 and includes minimally evaluation of 4 prognostically significant recurrent cytogenetic (CG) abnormalities (abn): deletion 13q (13q-), deletion 11q (11q-), trisomy 12 (+12) and 17p- as well as IGH abn in a pt subset. Complete clinical and pathological data has been systematically retrospectively collected on all pts who have had CLL FISH testing performed. We included all pts who had 32 sequential CLL FISH tests 33 months apart from 2004-Dec 2011. Primary endpoints were (a) incidence of CE and (b) overall survival.Of 861 pts screened, 134 met inclusion criteria. 21 pts (16%) had CE with median (med) age 57 yrs (range 45–80) at dx and 63 yrs (49–83) at time CE detected; 16 (77%) were male; Rai stage was low in 7 pts (33%), int in 11 (52%) and high in 3 (14%). At med follow-up of 7 yrs (1–37), 19/21 (91%) of CE pts and 99/113 (88%) non-CE pts received tx. Tx included nucleoside analog (NA) in 3 (14%) vs 23 (20%) pts, NA + rituximab (R) in 8 (38%) vs 53 (47%) pts, chlorambucil in 5 (24%) vs 14 (12%) pts and cyclophosphamide based in 3 (14%) vs 8 (7%) pts in the CE and non-CE groups respectively. 9 (43%) CE vs 36 (32%) non-CE pts underwent stem cell transplant (HSCT). 2 untx pts developed CE. CG abns detected on initial FISH test, classified by Dohner hierarchy were: 13q- (32%), +12 (22%), 17p- (12%), 11q- (11%), none of the 4 abns (23%). A total of 28 abns defining CE were detected in the 21pts and the most common were: 11q- (10 pts, 48%), 13q (10 pts, 47%). Predictors of CE on univariate analysis included lymphocyte count at dx (OR 1.01 per increase in lymphs by 1 ×109/L, 95% CI 1.00–1.02, p=0.048) and presence of 13q- at 1st FISH testing (OR 0.4, 95% CI 0.14–0.98, p=0.047); however these were not predictive of CE on multivariate analysis (MVA) after controlling for covariates (p=0.13 and p=0.19 respectively). No other baseline characteristics were predictive for CE including gender (p=0.31), age (p=0.38), WBC at dx (p=0.07), and CD38 positivity (p=0.56). OS was markedly reduced in pts who developed CE compared to those who did not: HR 5.9, median OS 8.2 yrs vs not reached, p=<0.001 (Fig 1). On MVA, CE retained its negative effect with HR 6.3 (95% CI 2.2–17.6), p<0.001, after adjusting for sex, age, Rai stage and baseline CG abn.CE is more common than previously recognized and carries a poor prognosis. In this study pts with CE fared worse than those with 17p-. OS of pts with CE in this cohort is in keeping with the med OS for 11q- and 17p- reported in the literature, indicating that presence of CE signifies a new poor prognostic indicator. These observations should prompt FISH testing in all pts at diagnosis and relevant time points in follow-up, such as with change in clinical behaviour and prior to new tx. Furthermore, as CE was thought to be tx related, the fact that 2 untx pts developed CE is a novel finding and provides further evidence of the clinical importance of CE in CLL. It is unclear whether CE pts should be considered for more aggressive tx, including HSCT. Results of our study provide a better understanding of the relationship of CE and disease progression in CLL, allowing for improved pt management and tx of this clinically heterogeneous and incurable disease.No relevant conflicts of interest to declare.