AIMS:External beam radiotherapy (EBRT) for prostate cancer (PCa) has rapidly advanced over the years. Advanced techniques with altered dose distributions may have an impact on second haematological cancer (SHC) risks. We assessed SHC risk after EBRT for PCa and explored whether this risk has changed over the years. MATERIALS AND METHODS:Patients diagnosed with a T1-T3 PCa between 1990 and 2015 were selected from the Netherlands Cancer Registry. Patients treated with EBRT were assigned to EBRT eras based on the date of diagnosis. These eras represented two-dimensional radiotherapy (2D-RT; 1991-1996), three-dimensional conformal radiotherapy (3D-CRT; 1998-2005) or advanced EBRT (2008-2015). Standardised incidence ratios (SIR) and absolute excess risks (AER) were calculated overall and by EBRT era. Sub-hazard ratios (sHRs) were calculated for the comparison of EBRT versus radical prostatectomy and active surveillance. RESULTS:PCa patients with EBRT as the primary treatment (n = 37 762) had an increased risk of developing a SHC (SIR = 1.20; 95% confidence interval 1.13-1.28) compared with the Dutch male general population. Estimated risks were highest for the 2D-RT era (SIR = 1.32; 95% confidence interval 1.14-1.67) compared with the 3D-CRT era (SIR = 1.16; 95% confidence interval 1.05-1.27) and the advanced EBRT era (SIR = 1.21; 95% confidence interval 1.07-1.36). AER were limited, with about five to six extra cases per 10 000 person-years. Relative risk analysis (EBRT versus radical prostatectomy/active surveillance) showed significant elevation with EBRT versus active surveillance (sHR = 1.17; 95% confidence interval 1.03-1.33; P = 0.017), but not for EBRT versus radical prostatectomy (sHR = 1.08; 95% confidence interval 0.94-1.23; P = 0.281). CONCLUSION:Increased SHC risks after EBRT for PCa cancer were observed for all EBRT eras compared with the general Dutch male population. Excess risks for EBRT versus other PCa treatment groups were found for only EBRT versus active surveillance.

Background Seeding of pathology related to Alzheimer's disease (AD) and Lewy body disease (LBD) by tissue homogenates or purified protein aggregates in various model systems has revealed prion-like properties of these disorders. Typically, these homogenates are injected into adult mice stereotaxically. Injection of brain lysates into newborn mice represents an alternative approach of delivering seeds that could direct the evolution of amyloid-β (Aβ) pathology co-mixed with either tau or α-synuclein (αSyn) pathology in susceptible mouse models. Methods Homogenates of human pre-frontal cortex were injected into the lateral ventricles of newborn (P0) mice expressing a mutant humanized amyloid precursor protein (APP), human P301L tau, human wild type αSyn, or combinations thereof. The homogenates were prepared from AD and AD/LBD cases displaying variable degrees of Aβ pathology and co-existing tau and αSyn deposits. Behavioral assessments of APP transgenic mice injected with AD brain lysates were conducted. For comparison, homogenates of aged APP transgenic mice that preferentially exhibit diffuse or cored deposits were similarly injected into the brains of newborn APP mice. Results We observed that lysates from the brains with AD (Aβ+, tau+), AD/LBD (Aβ+, tau+, αSyn+), or Pathological Aging (Aβ+, tau-, αSyn-) efficiently seeded diffuse Aβ deposits. Moderate seeding of cerebral amyloid angiopathy (CAA) was also observed. No animal of any genotype developed discernable tau or αSyn pathology. Performance in fear-conditioning cognitive tasks was not significantly altered in APP transgenic animals injected with AD brain lysates compared to nontransgenic controls. Homogenates prepared from aged APP transgenic mice with diffuse Aβ deposits induced similar deposits in APP host mice; whereas homogenates from APP mice with cored deposits induced similar cored deposits, albeit at a lower level. Conclusions These findings are consistent with the idea that diffuse Aβ pathology, which is a common feature of human AD, AD/LBD, and PA brains, may arise from a distinct strain of misfolded Aβ that is highly transmissible to newborn transgenic APP mice. Seeding of tau or αSyn comorbidities was inefficient in the models we used, indicating that additional methodological refinement will be needed to efficiently seed AD or AD/LBD mixed pathologies by injecting newborn mice.

Mouse models that replicate facets of human neurological diseases are often used at the pre-clinical stage to better understand the underlying mechanisms of a disease and test the target engagement of potential therapeutic interventions. We recently characterized a mouse model of childhood-onset parkinsonism-dystonia, a disease caused by a homozygous loss-of-function mutation in the SLC39A14 gene. The disease manifests itself phenotypically by impairments in locomotor behaviour and postural abnormalities. Our initial characterization of the model revealed that the Slc39a14-/- mice showed altered Mn homeostasis and compromised locomotor performance in vertical pole-descending, horizontal beam-traversing, and rotarod tests (Jenkitkasemwong et al., 2018). However, some of the mice also displayed torticollis and Straub tail. In this study, we investigated whether these postural abnormalities affected the performance in the above motility tests and consequently, biased and compromised the external validity of reported abnormal locomotor profiles. Our analyses showed that the Slc39a14-/- mice displaying torticollis and/or Straub tail had tests scores comparable to scores of their counterparts that never displayed these postural abnormalities. The z-score general index of performance revealed that the Slc39a14-/- model presents a complex pathological motor phenotype relevant to the complexity of phenotypes identified in childhood-onset parkinsonism-dystonia.

S92 ESTRO 38DVH parameters and optimization objectives were extracted from archived DVH reports.Data were analysed in SPSS. ResultsIn 10.7% (n=19) of cases the auto-plan was directly accepted for treatment.In 46.9% (n=83) of cases, MUs were scaled before accepting the auto-plan.In 40.1% (n=71), the auto-plan was optimised further.In 2.3% (n=4) of all cases, the auto-plan was rejected entirely and a new plan was made manually.We could identify the following reasons for manual adaptations: Bowel loop: 14.7% of plans (n=26), a bowel loop was near the PTV.In 4 cases MU were scaled and 22 cases were optimised further.

Hypothalamic-pituitary-adrenal (HPA) axis dysfunction contributes to numerous human diseases and disorders. We developed a high-affinity monoclonal antibody, CTRND05, targeting corticotropin-releasing factor (CRF). In mice, CTRND05 blocks stress-induced corticosterone increases, counteracts effects of chronic variable stress, and induces other phenotypes consistent with suppression of the HPA axis. CTRND05 induces skeletal muscle hypertrophy and increases lean body mass, effects not previously reported with small-molecule HPA-targeting pharmacologic agents. Multiorgan transcriptomics demonstrates broad HPA axis target engagement through altering levels of known HPA-responsive transcripts such as Fkbp5 and Myostatin and reveals novel HPA-responsive pathways such as the Apelin-Apelin receptor system. These studies demonstrate the therapeutic potential of CTRND05 as a suppressor of the HPA axis and serve as an exemplar of a potentially broader approach to target neuropeptides with immunotherapies, as both pharmacologic tools and novel therapeutics.

Understanding the biological functions of tau variants can illuminate differential etiologies of Alzheimer's disease (AD) and primary tauopathies. Though the end-stage neuropathological attributes of AD and primary tauopathies are similar, the etiology and behavioral outcomes of these diseases follow unique and divergent trajectories. To study the divergent physiological properties of tau variants on a uniform immunogenetic background, we created somatic transgenesis CNS models of tauopathy utilizing neonatal delivery of adeno-associated viruses expressing wild-type (WT) or mutant tau in non-transgenic mice. We selected four different tau variants-WT tau associated with AD, P301L mutant tau associated with frontotemporal dementia (FTD), S320F mutant tau associated with Pick's disease and a combinatorial approach using P301L/S320F mutant tau. CNS-targeted expression of WT and P301L mutant tau results in robust tau hyperphosphorylation without tangle pathology, gradually developing age-progressive memory deficits. In contrast, the S320F variant, especially in combination with P301L, produces an AD-type tangle pathology, focal neuroinflammation and memory impairment on an accelerated time scale. Using the doubly mutated P301L/S320F tau variant, we demonstrate that combining different mutations can have an additive effect on neuropathologies and associated co-morbidities, possibly hinting at involvement of unique functional pathways. Importantly, we also show that overexpression of wild-type tau as well as an FTD-associated tau variant can lead to cognitive deficits even in the absence of tangles. Together, our data highlights the synergistic neuropathologies and associated cognitive and synaptic alterations of the combinatorial tau variant leading to a robust model of tauopathy.

There is considerable interest in harnessing innate immunity to treat Alzheimer’s disease (AD). Here, we explore whether a decoy receptor strategy using the ectodomain of select TLRs has therapeutic potential in AD. AAV-mediated expression of human TLR5 ectodomain (sTLR5) alone or fused to human IgG4 Fc (sTLR5Fc) results in robust attenuation of amyloid β (Aβ) accumulation in a mouse model of Alzheimer-type Aβ pathology. sTLR5Fc binds to oligomeric and fibrillar Aβ with high affinity, forms complexes with Aβ, and blocks Aβ toxicity. Oligomeric and fibrillar Aβ modulates flagellin-mediated activation of human TLR5 but does not, by itself, activate TLR5 signaling. Genetic analysis shows that rare protein coding variants in human TLR5 may be associated with a reduced risk of AD. Further, transcriptome analysis shows altered TLR gene expression in human AD. Collectively, our data suggest that TLR5 decoy receptor–based biologics represent a novel and safe Aβ-selective class of biotherapy in AD.

Parkinson's disease manifests as a progressive movement disorder with underlying degeneration of dopaminergic neurons in the substantia nigra, consequent depletion of dopamine levels, and the accumulation of Lewy bodies in the brain. Because α-synuclein (α-Syn) protein is the major component of Lewy bodies, mouse models expressing wild-type or mutant SNCA/α-Syn genes provide a useful tool to investigate canonical characteristics of the disease. We evaluated a mouse model (denoted M20) that expresses human wild-type SNCA gene. The M20 mice showed abnormal locomotor behavior and reduced species-specific home cage activity. However, the direction of behavioral changes was task specific. In comparison with their control littermates, the M20 mice exhibited shorter grip endurance, and longer times to traverse elevated beams, but they descended the vertical pole faster and stayed longer on the accelerated rod than the control mice. The M20 mice were also impaired in burrowing and nest building activities. These results indicate a possible role of α-Syn in motor coordination and the motivation to perform species-specific behaviors in the presymptomatic model of synucleinopathy.

Intracellular accumulation of aggregated hyperphosphorylated tau protein is a neuropathological hallmark of neurodegenerative proteinopathies, such as Alzheimer's disease and primary tauopathies. Understanding how different tau variants lead to disease pathogenesis will allow us to mechanistically elucidate molecular properties and differential etiologies of wild type and tauopathy-associated variants. Currently there are several transgenic mouse models that can recapitulate substantial aspects of tauopathy. Though most of these faithfully recreate age-progressive hyperphosphorylation of tau, many do not show neurodegenerative changes, behavioral abnormalities or frank neurofibrillary tangles (NFT). While it is possible that these models may be tracking only the molecular properties of a particular tau variant, it is entirely possible that the strain of the mouse, its immune phenotype, the promoter used as well as the tissue distribution of the transgene can lead to these differential phenotypes. To allow us to study the molecular and physiological properties of tauopathy-associated tau variants on a standardized genetic background, we performed neonatal intraceberoventricular injections of recombinant adeno-associated viruses overexpressing different tau mutants in wild type mice. This results in, what we have previously described as, ‘somatic transgenesis’ models of tauopathy. Mice expressing different tau variants were aged and extensively characterized using behavioral and neuropathological parameters. We find that CNS targeted expression of wild type and several FTD-associated mutants (such as P301L and P301S) result in extensive age-progressive tau hyperphosphorylation. However, only the P301L tau expressing mice develop NFT by 9 months of age. In contrast, a Pick's disease associated variant leads to NFT by 3 months of age and a mouse model which expresses both P301L tau and the PiD variant leads to extensive neurodegenerative features, behavioral abnormalities and NFT by 3 months of age. We report various tauopathy models that allow us to molecularly phenotype different tauopathy-associated tau variants. These somatic transgenesis models allow us an alternative platform for cost-effective and rapid phenotyping of tau variants. These models can also be used to test different therapeutic interventions based on the particular molecular phenotype being targeted, i.e., tau phosphorylation, neurodegeneration, behavioral abnormalities and/or NFT.

Significance Manganese (Mn) is an essential nutrient that is toxic in excess. Exposure to excess Mn can result in Mn accumulation in the brain and neurological and motor disturbances resembling Parkinson disease. Here, we demonstrate that the transmembrane metal-ion transporter solute carrier family 39, member 14 (SLC39A14) is essential for Mn homeostasis. We provide evidence that SLC39A14 is required for efficient Mn uptake by the liver and pancreas, two organs that are known to actively participate in Mn excretion from the body. Accordingly, loss of SLC39A14 impairs Mn excretion, leading to Mn accumulation in the brain and most other extrahepatic tissues. Slc39a14 -deficient mice, similar to SLC39A14 -deficient human patients, display motor deficits, and thus offer a convenient model to study Mn/SLC39A14-related neurotoxicity.

Processing of amyloid-β (Aβ) precursor protein (APP) by γ-secretase produces multiple species of Aβ: Aβ40, short Aβ peptides (Aβ37–39), and longer Aβ peptides (Aβ42–43). γ-Secretase modulators, a class of Alzheimer’s disease therapeutics, reduce production of the pathogenic Aβ42 but increase the relative abundance of short Aβ peptides. To evaluate the pathological relevance of these peptides, we expressed Aβ36–40 and Aβ42–43 in Drosophila melanogaster to evaluate inherent toxicity and potential modulatory effects on Aβ42 toxicity. In contrast to Aβ42, the short Aβ peptides were not toxic and, when coexpressed with Aβ42, were protective in a dose-dependent fashion. In parallel, we explored the effects of recombinant adeno-associated virus–mediated expression of Aβ38 and Aβ40 in mice. When expressed in nontransgenic mice at levels sufficient to drive Aβ42 deposition, Aβ38 and Aβ40 did not deposit or cause behavioral alterations. These studies indicate that treatments that lower Aβ42 by raising the levels of short Aβ peptides could attenuate the toxic effects of Aβ42.

Epidemiologic and biomarker studies have posited an association between the experience of chronic psychologic stress and an increased incidence of Alzheimer's Disease (AD). Corticotropin-releasing hormone (CRH) and cortisol are two of the main mediators of the response to psychologic stress in humans, and both CRH and cortisol have been found to exacerbate amyloid ß deposition and tau phosphorylation in relevant mouse models. As AD patients have frequent comorbid behavioral and psychiatric dysfunctions including agitation and depression, stress signaling pathways may provide therapeutic targets that could simultaneously affect both behavioral status and protein deposition in AD patients. Historically CRH receptor (CRHR1) antagonists have failed in clinical trials for psychiatric disorders, and can potentially increase amyloid ß production through ß-arrestin mediated pathways. Therefore we aimed to develop a novel strategy for directly targeting CRH using a high affinity antibody. Using molecular cloning, vaccination, and hybridoma technology we have successfully generated and purified a monoclonal Anti CRH antibody for use as a potential therapeutic. This antibody is currently being tested as a therapeutic in both mutant APP and tau transgenic mice. A one time intraperitoneal injection of 25 mg/kg of Anti-CRH antibody in mice is able to provide >80% suppression of the corticosterone response to acute restraint stress and is able to reverse the cushingoid phenotype seen in mice that overexpress CRH within the brain. This suppression of corticosterone from one dose remains present even four days past injection and possibly much longer. We have generated a high affinity (Kd<1.0E-12) monoclonal CRH antibody that is able to reduce the glucocorticoid response to acute restraint stress by greater than 80% while also maintaining basal glucocorticoid levels in mice. These studies have broad implication for stress-related disorders in addition to AD, and may herald a new class of central nervous system therapies that target neuropeptides using high affinity antibodies.

MAPT mutations cause neurodegenerative diseases such as frontotemporal dementia but, strikingly, patients with the same mutation may have different clinical phenotypes.

Purpose: The risk of gastrointestinal cancer is increased in Hodgkin lymphoma (HL) survivors treated with abdominal radiation treatment (RT) and/or alkylating chemotherapy (CT). This study aims to evaluate whether survival of patients who survived HL and developed gastrointestinal cancer differs from survival of first primary gastrointestinal cancer patients. Patients and Methods: Overall and cause-specific survival rates of gastrointestinal (GI) cancer patients in a Dutch HL survivor cohort (n = 104, including esophageal, gastric, small intestinal, and colorectal cancer) were compared with survival of patients with a first primary GI cancer (n = 1025), generated through the Netherlands Cancer Registry by case matching on tumour site, gender, age, and year of diagnosis. Cox proportional hazards regression was used for survival analyses. Multivariable analyses were adjusted for GI cancer stage, grade of differentiation, surgery, RT, and CT. Results: Patients with GI cancers after HL were diagnosed at a median age of 54 years (interquartile range, 45-60). No differences in tumor stage or frequency of surgery were found between patients with GI cancer after HL and patients with first primary GI cancer. Patients with GI cancer after HL less often received RT (8% vs 23% in first primary GI cancer patients, P < .001) and CT (28% vs 41%, P = .01) for their GI tumor. Compared with patients with first primary GI cancer, overall and disease-specific survival rates of patients with gastrointestinal cancer after HL were worse (univariable hazard ratios [HRs] 1.30 [95% confidence interval (CI), 1.03-1.65], P = .03, and HR, 1.29 [95% CI, 1.00-1.67], P = .049, respectively). Adjustment for stage, grade, and treatment did not materially affect these risk estimates (HR, 1.33 [95% CI, 1.05-1.68], P = .02, and HR, 1.33 [95% CI 1.03-1.72], P = .03, respectively), indicating that the difference in survival between the 2 groups could not be explained by differences in stage, grade, or treatment. Mortality from other causes was slightly but not significantly higher in patients with GI cancer after HL compared with patients with first primary GI cancer (HR, 1.44 [95% CI, 0.81-2.56]; P = .22). Conclusion: Long-term overall and disease-specific survival of GI cancer in HL survivors are worse compared with first primary GI cancer patients. This survival difference, albeit not large, might result from a worse response of the tumour to treatment due to different pathogenesis of treatment-induced GI cancers. Keywords: Chemotherapy; Hodgkin lymphoma (HL)

43% P = .004) respectively. Table 1 highlights the strong association of both the level of education and SES with age in Brazilian patients. The educational level might influence directly the patient's capacity to recognize the potential graveness of lymphoma symptoms, as suggested by the longer time to diagnosis in illiterate individuals. Similarly, the level of education might impair the patient's ability to promptly react to complications of treatment. Moreover, the prevalence of comorbidities could differ according to the SES. Conclusions: Older Brazilian patients have advanced disease and non‐ nodular sclerosis subtype more often than younger patients. Treatment outcomes are inferior in older patients. These findings appear to be strongly associated with the lower educational level and lower SES observed in older patients.

Class II mice. These figures represent the counterparts of Fig. 5. stained with MC1, CP27, RZ3 and PHF1 antibodies, respectively. (ZIP 3613Â kb)

Pituitary Prolactin (PRL) and Growth Hormone (GH) are separately controlled and sub-serve different purposes. Surprisingly, we demonstrate that extra-pituitary expression in the adult mammalian central nervous system (CNS) is coordinated at mRNA and protein levels. However this was not a uniform effect within populations, such that wide inter-individual variation was superimposed on coordinate PRL/GH expression. Up to 44% of individuals in healthy cohorts of mice and rats showed protein levels above the norm and coordinated expression of PRL and GH transcripts above baseline occurred in the amygdala, frontal lobe and hippocampus of 10% of human subjects. High levels of PRL and GH present in post mortem tissue were often presaged by altered responses in fear conditioning and stress induced hyperthermia behavioral tests. Our data define a common phenotype polymorphism in healthy mammalian brains, and, given the pleiotropic effects known for circulating PRL and GH, further consequences of coordinated CNS over-expression may await discovery.