Although widely touted as a replacement for glass slides and microscopes in pathology, digital slides present major challenges in data storage, transmission, processing and interoperability. Since no universal data format is in widespread use for these images today, each vendor defines its own proprietary data formats, analysis tools, viewers and software libraries. This creates issues not only for pathologists, but also for interoperability. In this paper, we present the design and implementation of OpenSlide, a vendor-neutral C library for reading and manipulating digital slides of diverse vendor formats. The library is extensible and easily interfaced to various programming languages. An application written to the OpenSlide interface can transparently handle multiple vendor formats. OpenSlide is in use today by many academic and industrial organizations world-wide, including many research sites in the United States that are funded by the National Institutes of Health.

BackgroundOnly prototypes 5 years ago, high-speed, automated whole slide imaging (WSI) systems (also called digital slide systems, virtual microscopes or wide field imagers) are becoming increasingly capable and robust. Modern devices can capture a slide in 5 minutes at spatial sampling periods of less than 0.5 micron/pixel. The capacity to rapidly digitize large numbers of slides should eventually have a profound, positive impact on pathology. It is important, however, that pathologists validate these systems during development, not only to identify their limitations but to guide their evolution.MethodsThree pathologists fully signed out 25 cases representing 31 parts. The laboratory information system was used to simulate real-world sign-out conditions including entering a full diagnostic field and comment (when appropriate) and ordering special stains and recuts. For each case, discrepancies between diagnoses were documented by committee and a "consensus" report was formed and then compared with the microscope-based, sign-out report from the clinical archive.ResultsIn 17 of 25 cases there were no discrepancies between the individual study pathologist reports. In 8 of the remaining cases, there were 12 discrepancies, including 3 in which image quality could be at least partially implicated. When the WSI consensus diagnoses were compared with the original sign-out diagnoses, no significant discrepancies were found. Full text of the pathologist reports, the WSI consensus diagnoses, and the original sign-out diagnoses are available as an attachment to this publication.ConclusionThe results indicated that the image information contained in current whole slide images is sufficient for pathologists to make reliable diagnostic decisions and compose complex diagnostic reports. This is a very positive result; however, this does not mean that WSI is as good as a microscope. Virtually every slide had focal areas in which image quality (focus and dynamic range) was less than perfect. In some cases, there was evidence of over-compression and regions made "soft" by less than perfect focus. We expect systems will continue to get better, image quality and speed will continue to improve, but that further validation studies will be needed to guide development of this promising technology.

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Neoadjuvant HDI is highly effective for the treatment of palpable stage IIIB-C melanoma, and the findings of this study implicate an indirect immunomodulatory mechanism rather than a direct antitumor mechanism.

Defects in cholesterol synthesis result in a wide variety of symptoms, from neonatal lethality to the relatively mild dysmorphic features and developmental delay found in individuals with Smith-Lemli-Opitz syndrome. We report here the identification of mutations in sterol-C4-methyl oxidase-like gene (SC4MOL) as the cause of an autosomal recessive syndrome in a human patient with psoriasiform dermatitis, arthralgias, congenital cataracts, microcephaly, and developmental delay. This gene encodes a sterol-C4-methyl oxidase (SMO), which catalyzes demethylation of C4-methylsterols in the cholesterol synthesis pathway. C4-Methylsterols are meiosis-activating sterols (MASs). They exist at high concentrations in the testis and ovary and play roles in meiosis activation. In this study, we found that an accumulation of MASs in the patient led to cell overproliferation in both skin and blood. SMO deficiency also substantially altered immunocyte phenotype and in vitro function. MASs serve as ligands for liver X receptors α and β (LXRα and LXRβ), which are important in regulating not only lipid transport in the epidermis, but also innate and adaptive immunity. Deficiency of SMO represents a biochemical defect in the cholesterol synthesis pathway, the clinical spectrum of which remains to be defined. IntroductionCholesterol is a key component of cell membranes and lipid rafts and is the immediate precursor of steroids, vitamin D, and bile acids. Many disorders of cholesterol synthesis share common clinical features, such as abnormal morphogenesis, growth delay, and psychomotor disabilities (1). However, there are also striking differences suggesting that reduced de novo cholesterol synthesis per se may not primarily underlie some of the symptoms, including cataracts as well as skin and immune system abnormalities. Rather, recent studies implicate the accumulation of pre-cholesterol sterols and the replacement of cholesterol with some of these sterols in lipid rafts as playing a key role in the underlying pathophysiology (2). The meiosis-activating sterols (MASs) were the first group of cholesterol biogenesis intermediates that were found to have important extrahepatic functions in mammals. These include 4,4′-dimethyl-5α-cholesta-8,24-dien-3β-ol (testis meiosis-activating sterol [T-MAS]), 4,4′-dimethyl-5α-cholesta-8,14,24-trien-3β-ol (follicular fluid meiosis-activating sterols [FF-MASs]), and zymosterol. They are found in high concentration in testis and ovary and play roles in oocyte maturation and meiosis activation. The function of the MASs outside the reproductive organs is not well studied. FF-MAS is also a ligand for liver X receptors (LXRs) (3). LXR signaling is known to regulate crosstalk between inflammatory and cholesterol metabolism,

Purpose: The Janus-activated kinase/signal transducers and activators of transcription (STAT) pathway of IFN signaling is important to immunoregulation and tumor progression. STAT1plays a prominent role in the effector immune response, whereas STAT3 is implicated in tumor progression and down-regulation of the response to type IIFNs.The goal of this study was to understand the effects of high-dose IFNa2b (HDI) in relation to the balance of pSTAT1and pSTAT3. Experimental Design: We evaluated STAT1 and STAT3 jointly as mediators of IFNa effects in the setting of a prospective neoadjuvant trial of HDI, in which tissue samples were obtained before and after 20 doses of HDI therapy. Double immunohistochemistry for pSTAT1and pSTAT3 was done on paired fixed (9 patients) or frozen (12 patients) biopsies. Results: HDI was found to up-regulate pSTAT1, whereas it down-regulates pSTAT3 and total STAT3 levels in both tumor cells and lymphocytes. Higher pSTAT1/pSTAT3 ratios in tumor cells pretreatment were associated with longer overall survival (P = 0.032). The pSTAT1/pSTAT3 ratios were augmented by HDI both in melanoma cells (P = 0.005) and in lymphocytes (P = 0.022). Of the immunologic mediators and markers tested,TAP2 was augmented by HDI (but not TAP1and MHC class I/II). Conclusion: IFNa2b significantly modulates the balance of STAT1/STAT3 in tumor cells and host lymphocytes, leading to up-regulation of TAP2 and augmented host antitumor response. The pSTAT1/pSTAT3 ratio in tumor cells at baseline may serve as a useful predictor of clinical outcome in cutaneous melanoma; the modulation of this ratio may serve as a predictor of therapeutic effect.High-dose IFNa2b (HDI) is the only therapy that has shown a reproducible ability to prolong both relapse-free and overall survival of patients with resected high-risk, deep, primary, or lymph node metastatic melanoma. This therapy has consistently shown a capacity to reduce the hazard of relapse and mortality by 22% to 33% in multiple U.S. intergroup studies done over the past 20 years (1 -3). However, in the past decade, no further progress in the adjuvant therapy of melanoma has occurred, and the gap in our knowledge of the IFNa2b mechanism has proven to be a major impediment to further progress. A better understanding of the mechanism(s) of HDI in the adjuvant therapy of melanoma would enable more effective application of this therapy, and more intelligent strategies building upon this modality.The antitumor effects of IFNa2b include direct inhibition of tumor cell growth and the modulation of host immune response to tumor. The Janus-activated kinase/signal transducers and activators of transcription (STAT) signal pathway is one of the major mechanisms of IFNa2b action. Both human type I IFNa receptor chains 1 and 2 are required for the type I IFN -dependent signaling pathways. The intracellular domains of these receptors are associated with Janus-activated kinases, which are activated upon IFNa2b binding to its receptors. Janus-activated kinases then phosphorylate ...

BACKGROUND The management of pediatric melanoma (PM) has largely been extrapolated from adult data. However, the behavior of PM appears to differ from its adult counterparts. Therefore, an international PM registry was created and analyzed. METHODS Twelve institutions contributed deidentified clinicopathologic and outcome data for patients diagnosed with PM from 1953 through 2008. RESULTS Overall survival (OS) data were reported for 365 patients with invasive PM who had adequate follow-up data. The mean age of the patients was 16 years (range 1 year-21 years). The 10-year OS rate, 80.6%, tended to vary by patient age: 100% for those aged birth to 10 years, 69.7% for those aged > 10 years to 15 years, and 79.5% for those aged > 15 years to 20 years (P =.147). Patients with melanomas measuring ≤ 1 mm had a favorable prognosis (10-year OS rate of 97%), whereas survival was lower but similar for patients with melanomas measuring > 1 mm to 2 mm, > 2 mm to 4 mm, and > 4 mm (70%, 78%, and 80%, respectively; P =.0077). Ulceration and lymph node metastasis were found to be correlated with worse survival (P =.022 and P =.017, respectively). The 10-year OS rate was 94.1% for patients with American Joint Committee on Cancer stage I disease, 79.6% for those with stage II disease, and 77.1% for patients with stage III disease (P <.001). CONCLUSIONS Tumor thickness, ulceration, lymph node status, and stage were found to be significant predictors of survival in patients with PM, similar to adult melanoma. There is a trend toward increased survival in children aged ≤ 10 years versus adolescents aged > 10 years. Further analyses are needed to probe for potential biological and behavioral differences in pediatric versus adult melanoma.

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