The ALK tyrosine kinase inhibitor (TKI), crizotinib, shows significant activity in patients whose lung cancers harbor ALK fusions but its efficacy is limited by variable primary responses and acquired resistance. In work arising from the intriguing clinical observation of a patient with ALK fusion+ lung cancer who had an ‘exceptional response’ to an IGF-1R antibody, we define a therapeutic synergism between ALK and IGF-1R inhibitors. Similar to IGF-1R, ALK fusion proteins bind to the adaptor, IRS-1, and IRS-1 knockdown enhances the anti-tumor effects of ALK inhibitors. In models of ALK TKI resistance, the IGF-1R pathway is activated, and combined ALK/IGF-1R inhibition improves therapeutic efficacy. Consistent with this finding, IGF-1R/IRS-1 levels are increased in biopsy samples from patients progressing on crizotinib therapy. Collectively, these data support a role for the IGF-1R/IRS-1 pathway in both ALK TKI-sensitive and TKI-resistant states and provide biological rationale for further clinical development of dual ALK/IGF-1R inhibitors.

We describe a method of obtaining optical sectioning with a standard wide-field fluorescence microscope. The method involves acquiring two images, one with nonuniform illumination (in our case, speckle) and another with uniform illumination (in our case, randomized speckle). An evaluation of the local contrast in the speckle-illumination image provides an optically sectioned image with low resolution. This is complemented with high-resolution information obtained from the uniform-illumination image. A fusion of both images leads to a full resolution image that is optically sectioned across all spatial frequencies. This hybrid illumination method is fast, robust, and generalizable to a variety of illumination and imaging configurations.

We present a simple wide-field imaging technique, called HiLo microscopy, that is capable of producing optically sectioned images in real time, comparable in quality to confocal laser scanning microscopy. The technique is based on the fusion of two raw images, one acquired with speckle illumination and another with standard uniform illumination. The fusion can be numerically adjusted, using a single parameter, to produce optically sectioned images of varying thicknesses with the same raw data. Direct comparison between our HiLo microscope and a commercial confocal laser scanning microscope is made on the basis of sectioning strength and imaging performance. Specifically, we show that HiLo and confocal 3-D imaging of a GFP-labeled mouse brain hippocampus are comparable in quality. Moreover, HiLo microscopy is capable of faster, near video rate imaging over larger fields of view than attainable with standard confocal microscopes. The goal of this paper is to advertise the simplicity, robustness, and versatility of HiLo microscopy, which we highlight with in vivo imaging of common model organisms including planaria, C. elegans, and zebrafish.

The fundamentally stochastic nature of neuronal growth has hardly been addressed in neuroscience. We report on the stochastic fluctuations of a neuronal growth cone's leading edge movement, the basic step in neuronal growth. Describing the edge movement as a stochastic bistable process leads to an isotropic noise parameter that is successfully used to test the model. An analysis of growth cone motility confirms the model, and predicts that linear changes of the bistable potential, as known from stochastic filtering, result in directed growth cone translocation.

We present an endomicroscope apparatus that exhibits out-of-focus background rejection based on wide-field illumination through a flexible imaging fiber bundle. Our technique, called HiLo microscopy, involves acquiring two images, one with grid-pattern illumination and another with standard uniform illumination. An evaluation of the image contrast with grid-pattern illumination provides an optically sectioned image with low resolution. This is complemented with high-resolution information from the uniform illumination image, leading to a full-resolution image that is optically sectioned. HiLo endomicroscope movies are presented of fluorescently labeled rat colonic mucosa.

Sex cord-stromal tumours (SCSTs) consist of a heterogeneous group of neoplasms with diverse clinicopathological features and biological behaviour. They often present as a diagnostic challenge as they have varied and occasionally overlapping histomorphology and some may even mimic non-SCSTs. An accurate diagnosis is important for therapeutic and prognostic purposes. The use of a panel of immunohistochemical markers which are sensitive and specific for sex cord-stromal differentiation such as α-inhibin, calretinin, SF-1 and FOXL2, may be helpful in confirming the cellular lineage of these tumours, but is of limited utility in distinguishing between the different tumour types within this category. Additionally, the development of new therapeutic strategies in patients with SCSTs is also hampered by the infrequent occurrence of these neoplasms. Recent molecular analyses of some SCSTs has led to the discovery of novel molecular events, which may have important diagnostic, prognostic and therapeutic implications. The salient pathological features, management issues and recently described genetic aberrations in adult and juvenile granulosa cell tumours as well as Sertoli-Leydig cell tumours are discussed in this review, with particular emphasis on the clinical significance of FOXL2 and DICER1 mutations. An in-depth understanding of the molecular pathogenesis underlying SCSTs may aid in improving tumour classification and disease prognostication and also potentially lead to the discovery of more effective treatment strategies.

We describe a nonscanning, fiber bundle endomicroscope that performs optically sectioned fluorescence imaging with fast frame rates and real-time processing. Our sectioning technique is based on HiLo imaging, wherein two widefield images are acquired under uniform and structured illumination and numerically processed to reject out-of-focus background. This work is an improvement upon an earlier demonstration of widefield optical sectioning through a flexible fiber bundle. The improved device features lateral and axial resolutions of 2.6 and 17 μm, respectively, a net frame rate of 9.5 Hz obtained by real-time image processing with a graphics processing unit (GPU) and significantly reduced motion artifacts obtained by the use of a double-shutter camera. We demonstrate the performance of our system with optically sectioned images and videos of a fluorescently labeled chorioallantoic membrane (CAM) in the developing G. gallus embryo. HiLo endomicroscopy is a candidate technique for low-cost, high-speed clinical optical biopsies.

Aims Ovarian endometrioid carcinomas (OEC) of low grade have characteristic morphologic features, but high-grade tumors can mimic high-grade serous and undifferentiated carcinomas. We reviewed tumors initially diagnosed as OEC to determine whether a combination of pathologic and immunohistochemical features can improve histologic subclassification. Methods Tumors initially diagnosed as OEC were reviewed using World Health Organization criteria. We also noted the presence of associated confirmatory endometrioid features (CEFs): i) squamous metaplasia; ii) endometriosis; iii) adenofibromatous background; and iv) borderline endometrioid or mixed Mullerian component. A tissue microarray was constructed from 27 representative tumors with CEF and 14 without CEF, and sections were stained for WT-1, p16, and p53. Results Of 109 tumors initially diagnosed as OEC, 76 (70%) tumors were classified as OEC. The median patient age was 55 years and 75% of patients were younger than 60 years. 92% presented with disease confined to the pelvis and 87% of tumors were unilateral. The median tumor size was 11.8 cm. Only 3% of tumors were high-grade (grade 3 out of 3). 80% of cases had at least 1 CEF and 59% had at least 2 CEFs. 11% overexpressed p16, 0% overexpressed p53 and 3% expressed WT-1. Only 10% of patients died of disease at last follow-up. Thirty-three (33) tumors, or 30% of tumors originally classified as endometrioid, were re-classified as serous carcinoma (OSC). The median patient age was 54.5 years and 59% of patients were younger than 60 years of age. Only 27% had disease confined to the pelvis at presentation, 52% of tumors were unilateral and the median tumor size was 8 cm. Associated squamous differentiation, endometrioid adenofibroma and endometrioid or mixed Mullerian borderline tumor (CEFs) were not present in any case, but 6% of patients had endometriosis. Approximately one-half of the reclassified OSC demonstrated SET-pattern morphology (combinations of glandular, cribriform, solid and transitional cell-like architecture) and were immunophenotypically indistinguishable from OSCs with papillary architecture. 60% of OSC overexpressed p16, 50% overexpressed p53 and 82% expressed WT-1. At last follow-up, 52% had died of disease. Compared to OSC, OEC patients more frequently presented aged <60 years (p=0.046); had low-stage tumors (p<0.001); were more frequently unilateral (p<0.001); more frequent synchronous endometrial endometrioid carcinomas (p<0.001); and had no evidence of disease at last follow up (p<0.001). Their tumors were of lower grade (p<0.001); had more CEFs (p<0.001); and less frequently overexpressed p16 and p53 (p=0.003 and p<0.001, respectively) and less frequently expressed WT-1 (p<0.001). Conclusion This analysis emphasizes the diagnostic value of CEFs, the presence of a low-grade gland-forming endometrioid component and WT-1 negativity, as valid, clinically relevant criteria for a diagnosis of OEC. Glandular and/or cribriform architecture alone may be seen in both OECs and OSCs and are ther...