Validation of a zebrafish developmental defects assay as a qualified alternative test for its regulatory use following the ICH S5(R3) guideline.

Zebrafish is a popular toxicology model and provides an ethically acceptable small-scale analysis system with the complexity of a complete organism. Our goal is to further validate this model for its regulatory use for reproductive and developmental defects by testing the compounds indicated in the "Guideline on detection of reproductive and developmental toxicity for human pharmaceuticals" (ICH S5(R3) guideline.) To determine the embryotoxic and developmental risk of the 32 reference compounds listed in the ICH S5(R3) guideline, the presence of morphological alterations in zebrafish embryos was analyzed at two different stages to calculateLC50 and EC50 values for each stage. Teratogenic Indexes were established as the ratio between LC50 and EC50 critical for the proper compound classification as teratogenic when it is ≥ 2. A total of three biological replicates have been conducted to study the reproducibility of the assay. The chemicals' concentration in the medium and internally in the zebrafish embryos was evaluated. In this study, the 3 negative compounds were properly categorized while 23 compounds out of the 29 reference ones (sensitivity of 79.31%) were classified as teratogenic in zebrafish. The 6 that had false-negative results were classified 4 as inconclusive, 1 as not toxic, and 1 compound resulted toxic for zebrafish embryos under testing conditions. After the bioavailability experiments, some of the obtained inconclusive results were refined. The developmental defects assay in zebrafish gives an accuracy of 89.66%, sensitivity of 88.46%, specificity and repeatability of 100% compared to mammals; therefore, this is a well-integrated strategy using New Alternative Methods, to minimize the use of animals in developmental toxicity studies.

Efficient compressive and Bayesian characterization of biphoton frequency spectra.

Frequency-bin qudits constitute a promising tool for quantum information processing, but their high dimensionality can make for tedious characterization measurements. Here we introduce and compare compressive sensing and Bayesian mean estimation for recovering the spectral correlations of entangled photon pairs. Using a conventional compressive sensing algorithm, we reconstruct joint spectra with up to a 26-fold reduction in measurement time compared to the equivalent raster scan. Applying a custom Bayesian model to the same data, we then additionally realize reliable and consistent quantification of uncertainty. These efficient methods of biphoton characterization should advance our ability to use the high degree of parallelism and complexity afforded by frequency-bin encoding.

R-spondin 2 mediates neutrophil egress into the alveolar space through increased lung permeability.

OBJECTIVE: R-spondin 2 (RSPO2) is required for lung morphogenesis, activates Wnt signaling, and is upregulated in idiopathic lung fibrosis. Our objective was to investigate whether RSPO2 is similarly important in homeostasis of the adult lung. While investigating the characteristics of bronchoalveolar lavage in RSPO2-deficient (RSPO2-/-) mice, we observed unexpected changes in neutrophil homeostasis and vascular permeability when compared to control (RSPO2+/+) mice at baseline. Here we quantify these observations to explore how tonic RSPO2 expression impacts lung homeostasis. RESULTS: Quantitative PCR (qPCR) analysis demonstrated significantly elevated myeloperoxidase (MPO) expression in bronchoalveolar lavage fluid (BALF) cells from RSPO2-/- mice. Likewise, immunocytochemical (ICC) analysis demonstrated significantly more MPO+ cells in BALF from RSPO2-/- mice compared to controls, confirming the increase of infiltrated neutrophils. We then assessed lung permeability/barrier disruption via Fluorescein Isothiocyanate (FITC)-dextran instillation and found a significantly higher dextran concentration in the plasma of RSPO2-/- mice compared to identically treated RSPO2+/+ mice. These data demonstrate that RSPO2 may be crucial for blood-gas barrier integrity and can limit neutrophil migration from circulation into alveolar spaces associated with increased lung permeability and/or barrier disruption. This study indicates that additional research is needed to evaluate RSPO2 in scenarios characterized by pulmonary edema or neutrophilia.

Corticosteroid signaling at the brain-immune interface impedes coping with severe psychological stress.

The immune system supports brain plasticity and homeostasis, yet it is prone to changes following psychological stress. Thus, it remains unclear whether and how stress-induced immune alterations contribute to the development of mental pathologies. Here, we show that following severe stress in mice, leukocyte trafficking through the choroid plexus (CP), a compartment that mediates physiological immune-brain communication, is impaired. Blocking glucocorticoid receptor signaling, either systemically or locally through its genetic knockdown at the CP, facilitated the recruitment of Gata3- and Foxp3-expressing T cells to the brain and attenuated post-traumatic behavioral deficits. These findings functionally link post-traumatic stress behavior with elevated stress-related corticosteroid signaling at the brain-immune interface and suggest a novel therapeutic target to attenuate the consequences of severe psychological stress.

Contemporary management of men with high-risk localized prostate cancer in the United States.

This corrects the article DOI: 10.1038/pcan.2017.5.

Contemporary management of men with high-risk localized prostate cancer in the United States.

BACKGROUND: Surgery and radiation-based therapies are standard management options for men with clinically localized high-risk prostate cancer (PCa). Contemporary patterns of care are unknown. We hypothesize the use of surgery has steadily increased in more recent years. METHODS: Using the National Cancer Data Base for 2004-2013, all men diagnosed with high-risk localized PCa were identified using National Comprehensive Cancer Network criteria. Temporal trends in initial management were assessed. Multivariable logistic regression was used to evaluate demographic and clinical factors associated with undergoing radical prostatectomy (RP). RESULTS: In total, 127 391 men were identified. Use of RP increased from 26% in 2004 to 42% in 2013 (adjusted risk ratio (RR) 1.51, 95% CI 1.42-1.60, P<0.001), while external beam radiation therapy (EBRT) decreased from 49% to 42% (P<0.001). African American men had lower odds of undergoing RP (unadjusted rate of 28%, adjusted RR 0.69, 95% CI 0.66-0.72, <0.001) compared to White men (37%). Age was inversely associated with likelihood of receiving RP. Having private insurance was significantly associated with the increased use of RP (vs Medicare, adjusted odds ratio 1.04, 95% CI 1.01-1.08, P=0.015). Biopsy Gleason scores 8-10 with and without any primary Gleason 5 pattern were associated with decreased odds of RP (vs Gleason score ⩽6, both P<0.001). Academic and comprehensive cancer centers were more likely to perform RP compared to community hospitals (both P<0.001). CONCLUSION: The likelihood of receiving RP for high-risk PCa dramatically increased from 2004 to 2013. By 2013, the use of RP and EBRT were similar. African American men, elderly men and those without private insurance were less likely to receive RP.

Increasing incidence of metastatic prostate cancer in the United States (2004-2013).

BACKGROUND: Changes in prostate cancer screening practices in the United States have led to recent declines in overall incidence, but it is unknown whether relaxed screening has led to changes in the incidence of advanced and metastatic prostate cancer at diagnosis. METHODS: We identified all men diagnosed with prostate cancer in the National Cancer Data Base (2004-2013) at 1089 different health-care facilities in the United States. Joinpoint regressions were used to model annual percentage changes (APCs) in the incidence of prostate cancer based on stage relative to that of 2004. RESULTS: The annual incidence of metastatic prostate cancer increased from 2007 to 2013 (Joinpoint regression: APC: 7.1%, P<0.05) and in 2013 was 72% more than that of 2004. The incidence of low-risk prostate cancer decreased from years 2007 to 2013 (APC: -9.3%, P<0.05) to 37% less than that of 2004. The greatest increase in metastatic prostate cancer was seen in men aged 55-69 years (92% increase from 2004 to 2013). CONCLUSIONS: Beginning in 2007, the incidence of metastatic prostate cancer has increased especially among men in the age group thought most likely to benefit from definitive treatment for prostate cancer. These data highlight the continued need for nationwide refinements in prostate cancer screening and treatment.

Broadly tunable, low timing jitter, high repetition rate optoelectronic comb generator.

We investigate the low timing jitter properties of a tunable single-pass optoelectronic frequency comb generator. The scheme is flexible in that both the repetition rate and center frequency can be continuously tuned. When operated with 10 GHz comb spacing, the integrated residual pulse-to-pulse timing jitter is 11.35 fs (1 Hz to 10 MHz) with no feedback stabilization. The corresponding phase noise at 1 Hz offset from the photodetected 10 GHz carrier is -100 dBc/Hz.

Population-based analysis of treatment modalities and survival for clinically localized small-cell carcinoma of the prostate.

BACKGROUND: Small-cell carcinoma of the prostate is an aggressive cancer whose rarity has prevented the development of a consensus management approach. The objective of the current study was to determine the treatment patterns and evaluate factors affecting overall survival for patients with localized small-cell carcinoma of the prostate. METHODS: After querying the National Cancer Database, we identified all patients diagnosed with localized small-cell carcinoma of the prostate between 1998 and 2011 (n=287). Using Kaplan-Meier curves and Cox regression analyses, we assessed the effect of treatment and clinical stage on overall survival. RESULTS: Treatments included radiation therapy in 46% (n=131), chemotherapy in 38% (n=107), androgen deprivation therapy (ADT) in 22% (n=63) and radical prostatectomy in 13% (n=38). Median overall survival was 14.8 months. Upon multivariate analysis, local therapy (radical prostatectomy or radiation therapy) was associated with improved survival (hazard ratio (HR) 0.23, 95% confidence interval (CI) 0.14-0.38, P<0.001). Advanced clinical stage predicted worse survival among all men (cT3: HR 2.83, 95% CI 1.27-6.32, P=0.011; cT4: HR 3.26, 95% CI 1.50-7.07, P=0.003) and men who received local therapy (cT3: HR 4.67, 95% CI 1.41-15.44, P=0.012; cT4: HR 4.01, 95% CI 1.14-14.08, P=0.03) but not among men who received no local therapy (cT3: HR 1.64, 95% CI 0.51-5.27, P=0.4; cT4: HR 2.35, 95% CI 0.74-7.48, P=0.15). Age, receipt of chemotherapy and ADT, and clinical stage T2 disease (compared with T1) did not predict survival. CONCLUSION: Men with localized small-cell carcinoma of the prostate have a poor overall survival. Local therapy may represent a suitable and underused modality for select patients.

Role of polymorphic loci in HLA-region rs2647012 and rs805288 in the development of non-Hodgkin's malignant lymphomas in Western Siberia.

The data of genome-wide association analysis suggest that human 6p21.3 chromosomal region (localization of HLA genes) contains polymorphic loci influencing the risk of developing non-Hodgkin's lymphomas. We analyzed association of rs2647012 and rs805288 loci with the risk for non-Hodgkin's malignant lymphomas in the population of Western Siberia. Allele and genotype frequencies were determined in the group of 298 patients and in the control group including 551 individuals. Subgroups of diffuse large B-cell lymphoma (86 patients) and follicular lymphoma (25 patients) were analyzed separately. An association of rs2647012 А/А genotype with increased risk of the disease (OR = 2.78, p = 0.002) was detected in the subgroup of diffuse large B-cell lymphoma.

Bisphenol A affects larval growth and advances the onset of metamorphosis in Drosophila melanogaster.

Exposure to Bisphenol A (BPA) has been reported to dysregulate endocrine pathways in a wide array of vertebrate species. The effects of BPA on invertebrate species are less well understood. We tested the effects of BPA on growth and development in Drosophila as these processes are governed by well-studied endocrine pathways. In this study, we tested the effects of three concentrations of BPA (0.1mg/L, 1mg/L or 10mg/L) and found a statistically significant increase in larval growth for the low dose treatment group (0.1mg/L), but not statistically significant for the high dose treatment group (10mg/L). BPA exposure resulted in an increased body size in treated animals at 48, 72 and 96h after egg laying (AEL). This finding reflects a non-monotonic dose-response that has been observed for an increasing number of endocrine disrupting compounds. The increase in growth rate found for all treatment groups was associated with a statistically significant increase in food intake observed at 72h AEL. Furthermore, we observed that the increased growth rate was coupled with an earlier onset of pupariation consistent with previously reported phenotypes resulting from increased activity of insulin/insulin growth factor signaling (IIS) in Drosophila. Since the timing of the onset of pupariation in Drosophila is controlled through the complex interaction of the IIS and the ecdysone signaling pathways, our findings suggest that BPA exerts its effects through disruption of endocrine signaling in Drosophila.

Dural metastases in advanced prostate cancer: a case report and review of the literature.

Dural metastases from advanced prostate cancer are considered an uncommon diagnosis. However, autopsy studies show a high association between advanced prostate cancer and metastases to the meninges. Because the overall survival of advanced prostate cancer patients is expected to improve with the advent of new therapies, the incidence of clinically relevant dural metastases from prostate cancer will likely increase. We present a case of a heavily pre-treated castration-resistant prostate cancer patient who developed metastases to the duramater. This entity should be considered in the differential diagnosis of any patient with advanced castration-resistant prostate cancer and neurological symptoms. Clinicians should also be aware of the poor prognosis and survival rates associated with the condition.

Role of polymorphic variants of MTR gene A2756G and SHMT1 gene C1420T in the development of prostatic cancer in residents of the Western Siberian Region of Russia.

Allelic variants of folate cycle enzyme genes can contribute to predisposition to cancer. The impact of polymorphic loci A2756G of MTR gene and of C1420T of SHMT1 gene for the risk of prostatic cancer was studied in residents of West Siberia. The frequency of alleles of these loci in patients (N=371) and controls (N=285) was determined and the data were statistically processed. No statistically significant association with prostatic cancer was detected for any of the studied loci.

Detection and identification of banned processed animal protein in feedingstuffs by microscopic and PCR methods.

The aim of the study was to present the results of comparative evaluation of the usefulness of PCR and microscopic methods for the detection of Processed Animal Protein (PAP) in feedingstuffs. In the validation study, the limit of the detection for PCR was determined on 0.05% for beef, 0.1% for pork and 0.2% for poultry meat and bone meal (MBM). Among 62 doubtful samples of feedingstuffs examined by microscopic method 41 (66.13%) were found as positive. Based on the results obtained with the use of the microscopic and PCR methods it is possible to state that the molecular biology methods can, at present, be used as a supplementary method in PAP detection.

Dual-comb electric-field cross-correlation technique for optical arbitrary waveform characterization.

We present an electric-field cross-correlation technique that uses a pair of frequency combs to sweep phase and group delays independently without a mechanical stage. We demonstrate this technique for characterization of optical arbitrary waveforms composed of ~30 spectral lines from a 10 GHz frequency comb. Rapid data acquisition (tens of microseconds) enables interferometric spectral phase measurement of pulses subject to propagation over 20 km of optical fiber.

Generation of 20 GHz, sub-40 fs pulses at 960 nm via repetition-rate multiplication.

Optical filtering of a stabilized 1 GHz optical frequency comb produces a 20 GHz comb with approximately 40 nm bandwidth (FWHM) at 960 nm. Use of a low-finesse Fabry-Pérot cavity in a double-pass configuration provides a broad cavity coupling bandwidth (Deltalambda/lambda approximately 10%) and large suppression (50 dB) of unwanted modes. Pulse durations shorter than 40 fs with less than 2% residual amplitude modulation are achieved.

Improved signal-to-noise ratio of 10 GHz microwave signals generated with a mode-filtered femtosecond laser frequency comb.

We use a Fabry-Perot cavity to optically filter the output of a Ti:sapphire frequency comb to integer multiples of the original 1 GHz mode spacing. This effectively increases the pulse repetition rate, which is useful for several applications. In the case of low-noise microwave signal generation, such filtering leads to improved linearity of the high-speed photodiodes that detect the mode-locked laser pulse train. The result is significantly improved signal-to-noise ratio at the 10 GHz harmonic with the potential for a shot-noise limited single sideband phase noise floor near -168 dBc/Hz.

Interferometry from a scattering medium.

A two-beam random interferometer is demonstrated where coupling is facilitated by a scattering medium. A modulation observed in the normalized second-order intensity frequency correlation of the transmitted light is attributed to the relative temporal delay of the two beams and is insensitive to beam alignment and spacing.

CNBP mediates neural crest cell expansion by controlling cell proliferation and cell survival during rostral head development.

Striking conservation in various organisms suggests that cellular nucleic acid binding protein (CNBP) plays a fundamental biological role across different species. Recently, it was reported that CNBP is required for forebrain formation during chick and mouse embryogenesis. In this study, we have used the zebrafish model system to expand and contextualize the basic understanding of the molecular mechanisms of CNBP activity during vertebrate head development. We show that zebrafish cnbp is expressed in the anterior CNS in a similar fashion as has been observed in early chick and mouse embryos. Using antisense morpholino oligonucleotide knockdown assays, we show that CNBP depletion causes forebrain truncation while trunk development appears normal. A substantial reduction in cell proliferation and an increase in cell death were observed in the anterior regions of cnbp morphant embryos, mainly within the cnbp expression territory. In situ hybridization assays show that CNBP depletion does not affect CNS patterning while it does cause depletion of neural crest derivatives. Our data suggest an essential role for CNBP in mediating neural crest expansion by controlling proliferation and cell survival rather than via a cell fate switch during rostral head development. This possible role of CNBP may not only explain the craniofacial anomalies observed in zebrafish but also those reported for mice and chicken and, moreover, demonstrates that CNBP plays an essential and conserved role during vertebrate head development.