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BACKGROUND: Colonic atresias in the Fibroblast growth factor receptor 2IIIb (Fgfr2IIIb) mouse model have been attributed to increased epithelial apoptosis and decreased epithelial proliferation at embryonic day (E) 10.5. We therefore hypothesized that these processes would co-localize to the distal colon where atresias occur (atretic precursor) and would be excluded or minimized from the proximal colon and small intestine. RESULTS: We observed a global increase in intestinal epithelial apoptosis in Fgfr2IIIb(−/−) intestines from E9.5 to E10.5 that did not co-localize to the atretic precursor. Additionally, epithelial proliferations rates in Fgfr2IIIb(−/−) intestines were statistically indistinguishable to that of controls at E10.5 and E11.5. At E11.5 distal colonic epithelial cells in mutants failed to assume the expected pseudostratified columnar architecture and the continuity of the adjacent basal lamina was disrupted. Individual E-cadherin-positive cells were observed in the colonic mesenchyme. CONCLUSIONS: Our observations suggest that alterations in proliferation and apoptosis alone are insufficient to account for intestinal atresias and that these defects may arise from both a failure of distal colonic epithelial cells to develop normally and local disruptions in basal lamina architecture.

The National Institutes of Health leveled new focus on sex as a biological variable with the goal of understanding sex-specific differences in health and physiology. We previously published a functional assessment of the impact of sex, androgens, and prostate size on C57BL/6J mouse urinary physiology (Ruetten H, Wegner KA, Zhang HL, Wang P, Sandhu J, Sandhu S, Mueller B, Wang Z, Macoska J, Peterson RE, Bjorling DE, Ricke WA, Marker PC, Vezina CM. Am J Physiol Renal Physiol 317: F996–F1009, 2019). Here, we measured and compared five characteristics of urethral histology (urethral lumen diameter and area, epithelial cell count, epithelial and rhabdosphincter thickness, epithelial cell area, and total urethral area) in male and female 9-wk-old C57BL/6J mice using hematoxylin and eosin staining. We also compared male mice with castrated male mice, male and female mice treated with the steroid 5α-reductase inhibitor finasteride or testosterone, or male mice harboring alleles ( Pbsn4cre/+; R26RDta/+) that reduce prostate lobe mass. The three methods used to reduce prostate mass (castration, finasteride, and Pbsn4cre/+; R26RDta/+) changed urethral histology, but none feminized male urethral histology (increased urethral epithelial area). Exogenous testosterone caused increased epithelial cell count in intact females but did not masculinize female urethral histology (decrease epithelial area). Our results lay a critical foundation for future studies as we begin to parse out the influence of hormones and cellular morphology on male and female urinary function.

We genetically engineered expression of an activated form of P110 alpha, the catalytic subunit of PI3K, in mouse prostate epithelium to create a mouse model of direct PI3K activation (Pbsn-cre4Prb;PI3KGOF/+ ). We hypothesized that direct activation would cause rapid neoplasia and cancer progression. Pbsn-cre4Prb;PI3KGOF/+ mice developed widespread prostate intraepithelial hyperplasia, but stromal invasion was limited and overall progression was slower than anticipated. However, the model produced profound and progressive stromal remodeling prior to explicit epithelial neoplasia. Increased stromal cellularity and inflammatory infiltrate were evident as early as 4 months of age and progressively increased through 12 months of age, the terminal endpoint of this study. Prostatic collagen density and phosphorylated SMAD2-positive prostatic stromal cells were expansive and accumulated with age, consistent with pro-fibrotic TGF-β pathway activation. Few reported mouse models accumulate prostate-specific collagen to the degree observed in Pbsn-cre4Prb;PI3KGOF/+ . Our results indicate a signaling process beginning with prostatic epithelial PI3K and TGF-β signaling that drives prostatic stromal hypertrophy and collagen accumulation. These mice afford a unique opportunity to explore molecular mechanisms of prostatic collagen accumulation that is relevant to cancer progression, metastasis, inflammation and urinary dysfunction. © 2019 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Prostate autonomic and sensory axons control glandular growth, fluid secretion, and smooth muscle contraction and are remodeled during cancer and inflammation. Morphogenetic signaling pathways reawakened during disease progression may drive this axon remodeling. These pathways are linked to proliferative activities in prostate cancer and benign prostate hyperplasia. However, little is known about which developmental signaling pathways guide axon investment into prostate. The first step in defining these pathways is pinpointing when axon subtypes first appear in prostate. We accomplished this by immunohistochemically mapping three axon subtypes (noradrenergic, cholinergic, and peptidergic) during fetal, neonatal, and adult stages of mouse prostate development. We devised a method for peri-prostatic axon density quantification and tested whether innervation is uniform across the proximo-distal axis of dorsal and ventral adult mouse prostate. Many axons directly interact with or innervate neuroendocrine cells in other organs, so we examined whether sensory or autonomic axons innervate neuroendocrine cells in prostate. We first detected noradrenergic, cholinergic, and peptidergic axons in prostate at embryonic day (E) 14.5. Noradrenergic and cholinergic axon densities are uniform across the proximal-distal axis of adult mouse prostate while peptidergic axons are denser in the periurethral and proximal regions. Peptidergic and cholinergic axons are closely associated with prostate neuroendocrine cells whereas noradrenergic axons are not. These results provide a foundation for understanding mouse prostatic axon development and organization and, provide strategies for quantifying axons during progression of prostate disease.

Benign prostatic hyperplasia/lower urinary tract dysfunction (LUTD) affects nearly all men. Symptoms typically present in the fifth or sixth decade and progressively worsen over the remainder of life. Here, we identify a surprising origin of this disease that traces back to the intrauterine environment of the developing male, challenging paradigms about when this disease process begins. We delivered a single dose of a widespread environmental contaminant present in the serum of most Americans [2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD), 1 µg/kg], and representative of a broader class of environmental contaminants, to pregnant mice and observed an increase in the abundance of a neurotrophic factor, artemin, in the developing mouse prostate. Artemin is required for noradrenergic axon recruitment across multiple tissues, and TCDD rapidly increases prostatic noradrenergic axon density in the male fetus. The hyperinnervation persists into adulthood, when it is coupled to autonomic hyperactivity of prostatic smooth muscle and abnormal urinary function, including increased urinary frequency. We offer new evidence that prostate neuroanatomical development is malleable and that intrauterine chemical exposures can permanently reprogram prostate neuromuscular function to cause male LUTD in adulthood., Summary: We describe a new mechanism of benign prostate disease, initiated by fetal chemical exposure, which durably increases prostatic noradrenergic axon density and causes smooth muscle hyperactivity and urinary voiding dysfunction.

Anal squamous cell carcinoma (SCC) will be diagnosed in an estimated 9,080 adults in the United States this year, and rates have been rising over the last several decades. Most people that develop anal SCC have associated human papillomavirus (HPV) infection (∼85-95%), with approximately 5-15% of anal SCC cases occurring in HPV-negative patients from unknown etiology. This study identified and characterized a Kras-driven, female sex hormone-dependent development of anal squamous cell carcinoma (SCC) in the LSL-KrasG12D ; Pdx1-Cre (KC) mouse model that is not dependent on papillomavirus infection. One hundred percent of female KC mice develop anal SCC, while no male KC mice develop tumors. Both male and female KC anal tissue express Pdx1 and Cre-recombinase mRNA, and the activated mutant KrasG12D gene. Although the driver gene mutation KrasG12D is present in anus of both sexes, only female KC mice develop Kras-mutant induced anal SCC. To understand the sex-dependent differences, KC male mice were castrated and KC female mice were ovariectomized. Castrated KC males displayed an unchanged phenotype with no anal tumor formation. In contrast, ovariectomized KC females demonstrated a marked reduction in anal SCC development, with only 15% developing anal SCC. Finally, exogenous administration of estrogen rescued the tumor development in ovariectomized KC female mice and induced tumor development in castrated KC males. These results confirm that the anal SCC is estrogen mediated. The delineation of the role of female sex hormones in mediating mutant Kras to drive anal SCC pathogenesis highlights a subtype of anal SCC that is independent of papillomavirus infection. These findings may have clinical applicability for the papillomavirus-negative subset of anal SCC patients that typically respond poorly to standard of care chemoradiation.

The middle Miocene is marked by a changeover from a warm climatic period (Miocene Climatic Optimum, ~17-14.7 Ma), to a transitional phase (Middle Miocene Climatic Transition), culminating in a cold stage (Icehouse Mode, ~13.8 Ma). This period is associated with a positive excursion of δ13C (“Monterey Excursion”), showing a series of δ13C maxima (CM events) highlighted by cooling peaks in the δ18O values (Mi events). DSDP Site 372 obtained a high-resolution record of the middle Miocene of the western Mediterranean (Balearic Islands), and represents one of the best marine sedimentary records for the considered time interval. In this study, high-resolution stable isotope and quantitative calcareous plankton records for the Langhian to the early Serravallian time interval are presented. The stable oxygen and carbon isotope records allowed us to recognize and chronologically frame the Mi and CM events falling between 15.82 and 13.04 Ma. Furthermore, by integrating the stable isotope data with those obtained by the quantitative analyses of the calcareous plankton content, the three-folded climatic and environmental evolution was outlined for the considered time interval in the Mediterranean region. During the first stage (Miocene Climatic Optimum = MCO; from 17 up to 14.55 Ma), an initial warm-surface-water, oligotrophic, high salinity and restricted environment is followed by an open marine setting (starting from 15.18 Ma) within still warm-surface-water and oligotrophic conditions. The re-opening of the connections between the Mediterranean Sea and the Indian Ocean would have likely triggered this latter environmental change. From 14.55 to 13.75 Ma (second stage; Middle Miocene Climatic Transition; MMCT) an initial (up to 14.05 Ma) temperate-warm-surface-water and eutrophic environment occurred, followed by the onset of warm-surface-water, oligotrophic and slightly restricted marine conditions. The third stage (Icehouse Mode = IHM; from 13.75 Ma onwards) points to the development of a cold-water, eutrophic, high salinity environment in an open-marine setting.

The control of the response to tonal excitations or to broadband stochastic disturbances of a stiffened cylinder is investigated through the use of a Tuned Vibrating Absorber (TVA). In particular, the study considered both a purely passive device (Mechanical) and a semi-active one with shunt circuit (Electro-Mechanical) to evaluate the efficiencies and differences.

Urinary voiding dysfunction in aging men can cause bothersome symptoms and irreparable tissue damage. Underlying mechanisms are not fully known. We previously demonstrated that subcutaneous, slow-release testosterone and estradiol implants (T+E2) drive a pattern of urinary voiding dysfunction in male mice that resembles that of aging men. The initial goal of this study was to test the hypothesis that prostatic epithelial beta-catenin (Ctnnb1) is required for T+E2-mediated voiding dysfunction. Targeted Ctnnb1 deletion did not significantly change voiding function in control or T+E2 treated mice but led to the surprising discovery that the C57BL/6J × FVB/NJ × 129S1 mixed genetic background onto which Ctnnb1 loss of function alleles were maintained is profoundly susceptible to voiding dysfunction. The mixed background mice develop a more rapid T+E2-mediated increase in spontaneous urine spotting, are more impaired in ability to initiate bladder contraction, and develop larger and heavier bladders than T+E2 treated C57BL/6J pure bred mice. To better understand mechanisms, we separately evaluated contributions of T and E2 and found that E2 mediates voiding dysfunction. Our findings that genetic factors serve as modifiers of responsiveness to T and E2 demonstrate the need to control for genetic background in studies of male voiding dysfunction. We also show that genetic factors could control severity of voiding dysfunction. We demonstrate the importance of E2 as a key mediator of voiding impairment, and show that the concentration of E2 in subcutaneous implants determines the severity of voiding dysfunction in mice, demonstrating that the mouse model is tunable, a factor which is important for future pharmacological intervention studies.

The prostate develops by epithelial budding and branching processes that occur during fetal and postnatal stages. The adult prostate demonstrates remarkable regenerative capacity, with the ability to regrow to its original size over multiple cycles of castration and androgen administration. This capacity for controlled regeneration prompted the search for an androgen-independent epithelial progenitor in benign prostatic hyperplasia (BPH) and prostate cancer (PCa). BPH is hypothesized to be a reawakening of ductal branching, resulting in the formation of new proximal glands, all while androgen levels are decreasing in the aging male. Advanced prostate cancer can be slowed with androgen deprivation, but resistance eventually occurs, suggesting the existence of an androgen-independent progenitor. Recent studies indicate that there are multiple castration-insensitive epithelial cell types in the proximal area of the prostate, but not all act as progenitors during prostate development or regeneration. This review highlights how recent cellular and anatomical studies are changing our perspective on the identity of the prostate progenitor.

Background Castration-insensitive epithelial progenitors capable of regenerating the prostate have been proposed to be concentrated in the proximal region based on facultative assays. Functional characterization of prostate epithelial populations isolated with individual cell surface markers has failed to provide a consensus on the anatomical and transcriptional identity of proximal prostate progenitors. Methods Here, we use single-cell RNA sequencing to obtain a complete transcriptomic profile of all epithelial cells in the mouse prostate and urethra to objectively identify cellular subtypes. Pan-transcriptomic comparison to human prostate cell types identified a mouse equivalent of human urethral luminal cells, which highly expressed putative prostate progenitor markers. Validation of the urethral luminal cell cluster was performed using immunostaining and flow cytometry. Results Our data reveal that previously identified facultative progenitors marked by Trop2, Sca-1, KRT4, and PSCA are actually luminal epithelial cells of the urethra that extend into the proximal region of the prostate, and are resistant to castration-induced androgen deprivation. Mouse urethral luminal cells were identified to be the equivalent of previously identified human club and hillock cells that similarly extend into proximal prostate ducts. Benign prostatic hyperplasia (BPH) has long been considered an "embryonic reawakening," but the cellular origin of the hyperplastic growth concentrated in the periurethral region is unclear. We demonstrate an increase in urethral luminal cells within glandular nodules from BPH patients. Urethral luminal cells are further increased in patients treated with a 5-α reductase inhibitor. Conclusions Our data demonstrate that cells of the proximal prostate that express putative progenitor markers, and are enriched by castration in the proximal prostate, are urethral luminal cells and that these cells may play an important role in the etiology of human BPH.

Castration-insensitive epithelial progenitors capable of regenerating the prostate are concentrated in the proximal region close to the urethra, but the identification of these cells has been limited to individual cell surface markers. Here, we use single cell RNA sequencing (scRNA-seq) to obtain a cellular anatomy of the mouse prostate and urethra and create a comparative map with the human. These data reveal that previously identified facultative progenitors marked by TROP2, Sca-1, KRT4, and PSCA are actually luminal epithelia of the urethra that extend into the proximal prostate. These mouse urethral cells are the human equivalent of previously identified club and hillock urethral cells. Castration decreases androgen-dependent prostate luminal epithelia as expected, but TROP2+ urethral luminal epithelia survive and expand into the prostate. Benign prostatic hyperplasia (BPH) has long been considered an ‘embryonic reawakening’, but the cellular origin of peri-urethral growth is unclear. We use scRNA-seq and flow cytometry to demonstrate an increase in urethral luminal epithelia within glandular nodules from patients with BPH, which are further enriched in patients treated with a 5 alpha reductase inhibitor. These data demonstrate that the putative prostate progenitors enriched by castration in the proximal prostate are an expansion of urethral luminal epithelia and that these cells may play an important role in the etiology of human BPH.Significance StatementThe prostate involutes after castration, but regrows to its original size with androgen replenishment. This observation prompted the search for a castration-insensitive prostate progenitor. Here, Joseph et al. produce a comparative cellular atlas of the prostate and urethra in the mouse vs. human, discovering an equivalent urethral luminal epithelial cell type that extends into the proximal prostatic ducts and expresses previously identified markers of facultative prostate progenitors. Urethral luminal epithelia are established before prostate budding in human and mouse development, and expand after castration in the mouse and after 5 alpha reductase inhibitor treatment in human BPH. These data suggest that luminal epithelia of the urethra are castration-insensitive cells of proximal ducts that may act as progenitors in human BPH.

One-atom-thick rare-earth/noble metal (RE-NM) compounds are attractive materials to investigate two-dimensional magnetism, since they are easy to synthesize into a common RE-NM2 structure with high crystal perfection. Here we perform a comparative study of the GdAu2, HoAu2, and YbAu2 monolayer compounds grown on Au(111). We find the same atomic lattice quality and moiré superlattice periodicity in the three cases, but different electronic properties and magnetism. The YbAu2 monolayer reveals the characteristic electronic signatures of a mixed-valence configuration in the Yb atom. In contrast, GdAu2 and HoAu2 show the trivalent character of the rare-earth and ferromagnetic transitions below 22 K. Yet, the GdAu2 monolayer has an in-plane magnetic easy-axis, versus the out-of-plane one in HoAu2. The electronic bands of the two trivalent compounds are very similar, while the divalent YbAu2 monolayer exhibits different band features. In the latter, a strong 4f–5d hybridization is manifested in neatly resolved avoided crossings near the Fermi level. First principles theory points to a residual presence of empty 4f states, explaining the fluctuating valence of Yb in the YbAu2 monolayer., This work was supported in part by the Spanish MINECO (MAT-2017-88374-P, MAT2016-78293-C6, FIS2016-75862-P, PGC2018-098613-B-C21, SEV-2017-0706 and SEV-2016-0686), the Spanish Research Agency (PID2019-107338RB-C65), Basque Government Project IT-1255-19, and University of the Basque Country (UPV/EHU) grant GIU18/138 and the European Regional Development Fund (ERDF) under the program Interreg V-A España-Francia-Andorra (Contract No. EFA 194/16 TNSI). The experiments at Elettra were supported by the EU Calipso project and computational resources were provided by the DIPC computing center. L. F. acknowledges financial support from the European Union's Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement MagicFACE No 797109. A. K. K., M. J., P. M. S. and P. M. acknowledge the project EUROFEL-ROADMAP ESFRI.

Correction for ‘Influence of 4f filling on electronic and magnetic properties of rare earth-Au surface compounds’ by L. Fernandez et al., Nanoscale, 2020, 12, 22258–22267, DOI: 10.1039/D0NR04964F.

In RPA V-VI glioblastoma patients both hypofractionated radiotherapy and exclusive temozolomide can be used; the purpose of this trial is to compare these treatment regimens in terms of survival and quality of life. Patients with histologic diagnosis of glioblastoma were randomized to hypofractionated radiotherapy (RT–30 Gy in 6 fractions) and exclusive chemotherapy (CHT–emozolomide 200 mg/m2/day 5 days every 28 days). Overall (OS) and progression free survival (PFS) were evaluated with Kaplan Maier curves and correlated with prognostic factors. Quality- adjusted survival (QaS) was evaluated according to the Murray model (Neurological Sign and Symptoms–NSS) From 2010 to 2015, 31 pts were enrolled (CHT: 17 pts; RT: 14pts). Four pts were excluded from the analysis. RPA VI (p = 0.048) and absence of MGMT methylation (p = 0.001) worsened OS significantly. Biopsy (p = 0.048), RPA class VI (p = 0.04) and chemotherapy (p = 0.007) worsened PFS. In the two arms the initial NSS scores were overlapping (CHT: 12.23 and RT: 12.30) and progressively decreased in both group and became significantly worse after 5 months in CHT arm (p = 0.05). Median QaS was 104 days and was significantly better in RT arm (p = 0.01). The data obtained are limited by the poor accrual. Both treatments were well tolerated. Patients in RT arm have a better PFS and QaS, without significant differences in OS. The deterioration of the NSS score would seem an important parameter and coincide with disease progression rather than with the toxicity of the treatment.

Experimental evidence shows that soils subjected to dynamic excitations have both the ability to store and to dissipate strain energy. The phenomenon of energy dissipation takes place even at very small strain levels, below the linear cyclic threshold shear strain. From a phenomenological point of view this type of material behaviour can conveniently be described by the linear theory of viscoelasticity. An important result predicted by this theory is the functional dependence between the velocity of the propagation of body waves and the material damping ratio. Hence, in contrast to usual practice, these parameters should be measured simultaneously and at the same frequency of excitation. In this article the authors present a new experimental procedure to be conducted in laboratory with resonant column apparatus where the shear wave velocity and the shear damping ratio are measured simultaneously. Since these parameters are determined at specific frequencies of excitation, the method is also well suited to investigate the frequency dependence laws of these important soil parameters.

Non-invasive tests to identify age and early disease-associated pathology within the kidney are needed. Specific populations of urinary extracellular vesicles (EVs) could potentially be used for such a diagnostic test. Random urine samples were obtained from age- and sex-stratified living kidney donors before kidney donation. A biopsy of the donor kidney was obtained at the time of transplantation to identify nephron hypertrophy (larger glomerular volume, cortex per glomerulus and mean profile tubular area) and nephrosclerosis (% fibrosis, % glomerulosclerosis and arteriosclerosis). Renal parenchymal-derived EVs in cell-free urine were quantified by digital flow cytometry. The relationship between these EV populations and structural pathology on the kidney biopsy was assessed. Clinical characteristics of the kidney donors (n=138, age range: 20–70 years, 50% women) were within the normative range. Overall, urine from women contained more EVs than that from men. The number of exosomes, juxtaglomerular cells and podocyte marker–positive EVs decreased (p

New magnetobiostratigraphic data for the late Oligocene through early Miocene at Deep Sea Drilling Project (DSDP) Hole 516F provide a significantly revised age model, which permits reevaluation of developments that led to the Mi-1 glacial event at the Oligocene-Miocene boundary. Our new high-resolution paleomagnetic study, which is supported by quantitative calcareous nannofossil and planktonic foraminiferal analyses, significantly refines previous age models for Oligocene-Miocene sediments from DSDP Hole 516F, with ages that are systematically younger than those previously determined. In some parts of the Oligocene, the discrepancy with previous studies exceeds 450 kyr. Based on this new age model, we infer a progressive increase in sedimentation rate and paleoproductivity between circa 23.9 Ma and circa 22.9 Ma, with the highest rate coinciding with the Mi-1 glacial event at the Oligocene-Miocene boundary. This productivity increase would have resulted in higher rates of carbon burial and in turn a drawdown of atmospheric CO2. Immediately afterward, an abrupt decrease in sedimentation rate and paleoproductivity suggests that the Mi-1 deglaciation was associated with decreased carbon input into the ocean. Elevated sedimentation rates are also documented at ~24.5 Ma, coincident with the Oi2D glacioeustatic event. The presence of volcanic material within the sediments during these glacial events is interpreted to have resulted from redeposition of sediment scoured from nearby sites on the Rio Grande Rise due to transient variations in bottom water flow patterns.

After the formalization of the base of the Miocene in the Lemme-Carrosio section (Italy) at the base of Subchron C6Cn.2n, the calcareous plankton biostratigraphy was refined in several open ocean Deep Sea Drilling Project/Ocean Drilling Program sites. However, high-resolution quantitative biostratigraphic studies, integrating planktonic foraminifera and calcareous nannofossils, are still lacking for the time interval spanning the Oligocene–Miocene transition. Here, we present a reinvestigation of Deep Sea Drilling Project Hole 516F (Rio Grande Rise) and 4 oil wells drilled by Petrobras Brasileiro SA in the Campos Basin (SW Atlantic Ocean). We identified 12 planktonic foraminiferal and 18 calcareous nannofossil bioevents that have been integrated with an updated magnetostratigraphy of Hole 516F allowing the correlation with the GPTS and the identification of the Oligocene/Miocene boundary (base of Subchron C6Cn.2n) between the Top of Sphenolithus delphix and the Base of common Paragloborotalia kugleri. Furthermore, our results give new insights on the reliability of major calcareous plankton events across the Oligocene–Miocene transition: (a) the Sphenolithus ciperoensis Top, the S. delphix Base and Top, and the Sphenolithus cometa Base are reliable events at a global scale; (b) the Bases of Globoquadrina dehiscens and Sphenolithus disbelemnos > 4 μm are correlatable events only within the study sector of the SW Atlantic Ocean; and (c) the Globoturborotalita ciperoensis Top, Globoturborotalita angulisuturalis Top, and Sphenolithus procerus Base are diachronous. Finally, previously unreported biostratigraphic data, such as the distribution range of S. disbelemnos

On the wake of the results obtained so far at the SPARC_LAB test-facility at the Laboratori Nazionali di Frascati (Italy), we are currently investigating the possibility to design and build a new multi-disciplinary user-facility, equipped with a soft X-ray Free Electron Laser (FEL) driven by a ∼ 1 GeV high brightness linac based on plasma accelerator modules. This design study is performed in synergy with the EuPRAXIA design study. In this paper we report about the recent progresses in the on going design study of the new facility. On the wake of the results obtained so far at the SPARC\_LAB test-facility at the Laboratori Nazionali di Frascati (Italy), we are currently investigating the possibility to design and build a new multi-disciplinary user-facility, equipped with a soft X-ray Free Electron Laser (FEL) driven by a $\sim$1 GeV high brightness linac based on plasma accelerator modules. This design study is performed in synergy with the EuPRAXIA design study. In this paper we report about the recent progresses in the on going design study of the new facility.

Though many methods can be used to identify cell types contained in complex tissues, most require cell disaggregation and destroy information about where cells reside in relation to their microenvironment. Here, we describe a polytomous key for cell type identification in intact sections of adult mouse prostate and prostatic urethra. The key is organized as a decision tree and initiates with one round of immunostaining for nerve, epithelial, fibromuscular/hematolymphoid, or vascular associated cells. Cell identities are recursively eliminated by subsequent staining events until the remaining pool of potential cell types can be distinguished by direct comparison to other cells. We validated our identification key using wild type adult mouse prostate and urethra tissue sections and it currently resolves sixteen distinct cell populations which include three nerve fiber types as well as four epithelial, five fibromuscular/hematolymphoid, one nerve-associated, and three vascular-associated cell types. We demonstrate two uses of this novel identification methodology. We first used the identification key to characterize prostate stromal cell type changes in response to constitutive phosphatidylinositide-3-kinase activation in prostate epithelium. We then used the key to map cell lineages in a new reporter mouse strain driven by Wnt10aem1(cre/ERT2)Amc. The identification key facilitates rigorous and reproducible cell identification in prostate tissue sections and can be expanded to resolve additional cell types as new antibodies and other resources become available.

The Vicchio outcrop in the Tuscan Apennines contains excellent exposures of a Miocene methane-derived carbonate system, made up of more than 80 carbonate bodies enclosed in marly sediments. Facies analysis, composition and a detailed biostratigraphic study of the carbonates and enclosing Vicchio Marls allowed us to document the role of climatic changes and eustasy on seepage in these ancient deposits. Results of our study indicate that the stratigraphic horizon bearing seep-carbonates is constrained by two planktonic foraminiferal events, the Acme End (AE) of Turborotalita cf. T. quinqueloba (13.75 Ma) and the Acme 1 Beginning (A 1 B) of Paragloborotalia siakensis (13.32 Ma), encompassing about 400,000 years. The AE of T. cf. T. quinqueloba approximates the mid-Miocene global cooling event (Mi3b), as defined by the δ 18 O maxima (13.78 Ma). The glacio-eustatic sea level drop associated with this cooling event is estimated to be approximately 60 m. Assuming that the highest rates of glacio-eustatic sea level fall coincide with δ 18 O maxima, the timing of the seepage onset (at about 13.75 Ma) is concomitant with the phase of sea level-lowering. The ascent and emission of methane-rich fluids may have been triggered by pressure drop due to the eustatic fall associated with the Mi3b event. The moderate intensity of fluid expulsion is suggested by the lack of brecciation, by the scarcity of detrital particles in the carbonates and by the pervasive occurrence of carbonate in the enclosing marls. This type of seepage differs from focused fault-confined fluid flows, typical of other tectonically controlled Apenninic seep-carbonates.

Excellent exposures of Miocene seep-carbonates enclosed in marine marly sediments (Vicchio outcrops in the northern Apennines, Italy) offer the opportunity to highlight the evolution of a fossil seep ecosystem and the response of benthic communities to high-stressed environmental conditions. For this purpose, seep-related facies and molluscs within carbonate bodies have been studied, coupled with benthic foraminiferal assemblages and carbon and oxygen isotopes in the enclosing marls. The integrated planktonic foraminiferal and calcareous nannofossil biostratigraphy has allowed us to constrain the seepage within well-calibrated bioevents and to stress the relationships with paleoclimatic and paleoceanographic variations during the middle Miocene in the Mediterranean area. Our biostratigraphic data indicate that the onset of the seepage approximates the Mi3b cooling event (13.82 Ma) and the seepage system lasts for 400 kyr. The evolution of the Vicchio cold-seep system passes through four phases: (1) The onset of the seepage, characterized by a pervasive flow of methane-rich fluids, is inferred by δ13C depletion of marly sediments and by prevailing benthic foraminifera indicative of suboxic conditions at the sea-floor. (2) The methane flow becomes focused causing the precipitation of wide pinnacle-like carbonate bodies which contain giants lucinids. Enclosing marls indicate well-oxygenated conditions, possibly enhanced by paleoceanographic variations connected to the Mi3b cooling event. (3) The appearance of the vesicomyid Christineconcha cf. C. regab and the absence of lucinids in seep-carbonates suggest stable methane-rich fluid emissions; higher flow rates locally favoured the flourishment of bacterial mats. Benthic foraminifera show abundance peaks of organic matter depending taxa. (4) The reduced intensity of methane-rich fluid flows favours the precipitation of stratiform carbonate bodies along strike; the macrofauna is characterized by the presence of both Vesicomyidae and giant lucinids. Foraminiferal assemblages in the enclosing marls indicate the restoration of well-oxygenated conditions.

Juglans nigra and Juglans regia are two highly economically important species for wood and fruit production that are susceptible to anthracnose caused by Gnomonia leptostyla. The identification of genotypes resistant to anthracnose could represent a valid alternative to agronomic and chemical management. In this study, we analyzed 72 walnut genotypes that showed a variety of resistance phenotypes in response to natural infection. According to the disease severity rating and microsatellite fingerprinting analysis, these genotypes were divided into three main groups: (40) J. nigra resistant, (1) J. nigra susceptible, and (31) J. regia susceptible. Data on leaf emergence rates and analysis of in vivo pathogenicity indicated that the incidence of anthracnose disease in the field might be partially conditioned by two key factors: the age and/or availability of susceptible leaves during the primary infection of fungus (avoidance by late flushing) and partial host resistance. NBS profiling approach, based on PCR amplification with an adapter primer for an adapter matching a restriction enzyme site and a degenerate primer targeting the conserved motifs present in the NBS domain of NBS-LRR genes, was applied. The results revealed the presence of a candidate marker that correlated to a reduction in anthracnose incidence in 72 walnut genotypes.

article i nfo Continuous marine successions covering the Middle Miocene Climate Transition (MMCT; ∼15-13.7 Ma) are scarce and the lack of a high-resolution magnetobiostratigraphic framework hampers the construction of astronomically tuned age models for this time interval. The La Vedova High Cliff section, exposed along the coast of the Conero Riviera near Ancona (Italy), is one of the few Mediterranean sections covering the critical time interval of the MMCT. Starting from an initial magnetobiostratigraphic age model, a robust astronomical tuning was constructed for the interval between 14.2 and 13.5 Ma, using geochemical element data and time series analysis. A shift in δ 18 O of bulk sediment towards heavier values occurs between ∼13.92 and 13.78 Ma and could be related to the Mi3b oxygen isotope event, which reflects the rapid expansion of the East Antarctic Ice Sheet in the middle Miocene. The onset of the CM6 carbon excursion is reflected in the bulk record by a rapid increase in δ 13 C at 13.86 Ma. Our results confirm the proposition that these events coincide with a 405-kyr minimum in eccentricity and a node in obliquity related to the ∼1.2 Myr cycle. From 13.8 Ma onwards, distinct quadruplet cycles containing sapropelitic sediments were deposited. This may suggest a causal connection between the main middle Miocene cooling step and the onset of sapropel formation in the

The Global Stratotype Section and Point (GSSP) for the Base of the Serravallian Stage (Middle Miocene) is defined in the Ras il Pellegrin section located in the coastal cliffs along the Fomm Ir-Rih Bay on the west coast of Malta (35°54'50"N, 14°20'10"E). The GSSP is at the base of the Blue Clay Formation (i.e., top of the transitional bed of the uppermost Globigerina Limestone). This boundary between the Langhian and Serravallian stages coincides with the end of the major Mi-3b global cooling step in the oxygen isotopes and reflects a major increase in Antarctic ice volume, marking the end of the Middle Miocene climate transition and the Earth's transformation into an "Icehouse" climate state. The associated major glacio-eustatic sea-level drop corresponds with sequence boundary Ser1 of Hardenbol et al. (1998) and supposedly with the TB2.5 sequence boundary of Haq et al (1987). This event is slightly older than the last common and/or continuous occurrence of the calcareous nannofossil Sphenolithus heteromorphus, previously considered as guiding criterion for the boundary, and is projected to fall within the younger half of Chron C5ACn. The GSSP level is in full agreement with the definitions of the Langhian and Serravallian in their respective historical stratotype sections in northern Italy and has an astronomical age of 13.82 Ma.

An integrated magneto-biostratigraphic framework is presented for Middle Miocene sediments of DSDP Site 372 located in the Western Mediterranean. Detailed biostratigraphic analysis shows a nearly complete sequence of early Middle Miocene calcareous plankton bioevents in the Mediterranean, including the LCO (Last Common Occurrence) of the nannofossil Sphenolithus heteromorphus which has been astronomically dated in the Ras il Pellegrin (RIP) section on Malta Island [Abels, H.A., Hilgen, F.J., Krijgsman, W., Kruk, R.W., Raffi, I., Turco, E., Zachariasse, W.J., 2005. Long-period orbital control on middle Miocene global cooling: integrated stratigraphy and astronomical tuning of the Blue Clay Formation on Malta, Paleoceanography, 20, PA4012. doi: 10.1029/2004PA001129. 11 pp]. Thermal demagnetization of discrete samples revealed a characteristic low-temperature component with dual polarities despite a weak paleomagnetic signal. The resultant magnetostratigraphic record, combined with the calcareous plankton biostratigraphy, is straightforwardly correlated to the geomagnetic polarity time scale (CK95) of Cande and Kent [Cande, S.C., Kent, D.V., 1995. Revised calibration of the Geomagnetic Polarity Time Scale for the Late Cretaceous and Cenozoic, J. Geophys. Res., 100, 6093-6095] and the Astronomical Tuned Neogene Time Scale (ATNTS04) of Lourens et al. [Lourens, L.J., Hilgen, F.J., Laskar, J., Shackleton, N.J., Wilson, D., 2004. The Neogene Period. In: Gradstein F.M., Ogg J.G., Smith A.G. (Eds.), A Geologic Time Scale, Cambridge Univ. Press, pp. 409-440]. The subchrons recorded in Site 372 succession range from C5Br up to C5AAr. To confirm the magnetostratigraphic calibration, 40Ar/39Ar dating was performed on feldspar of two volcanic ash layers. The radio-isotopic dating indicates a younger age for these two ash layers compared to the magnetostratigraphic calibrated ages according to the CK95 and ATNTS04 age models. However, if the astronomically calibrated age of 28.21 ± 0.04 Ma is used for the Fish Canyon standard (FCs), the age for the older ash layer exactly matches its ATNTS04 age. Ages for bioevents were calculated assuming constant sedimentation rates between magnetostratigraphic age-tie points. The S. heteromorphus LCO has an age of 13.54 Ma and 13.63 Ma according to CK95 and ATNTS04, respectively, which is consistent with the astronomical tuned age of 13.65 Ma determined at RIP section [Abels, H.A., Hilgen, F.J., Krijgsman, W., Kruk, R.W., Raffi, I., Turco, E., Zachariasse, W.J., 2005. Long-period orbital control on middle Miocene global cooling: integrated stratigraphy and astronomical tuning of the Blue Clay Formation on Malta, Paleoceanography, 20, PA4012. doi:10.1029/2004PA001129. 11 pp]. We therefore conclude that the magnetostratigraphic calibration of DSDP Site 372 is correct and that, for the time being, this site can be considered as a reference section for the early Middle Miocene of the Mediterranean region.

A revised stratigraphic framework for the Messinian succession of Cyprus is proposed demonstrating that the three-stage model for the Messinian salinity crisis recently established for the Western Mediterranean also applies to the Eastern Mediterranean, at least for its marginal basins. This analysis is based on a multidisciplinary study of the Messinian evaporites and associated deposits exposed in the Polemi, Pissouri, Maroni/Psematismenos and Mesaoria basins. Here, we document for the first time that the base of the unit usually referred to the ‘Lower Evaporites’ in Cyprus does not actually correspond to the onset of the Messinian salinity crisis. The basal surface of this unit rather corresponds to a regional-scale unconformity, locally associated with an angular discordance, and is related to the erosion and resedimentation of primary evaporites deposited during the first stage of the Messinian salinity crisis. This evidence suggests that the ‘Lower Evaporites’ of the southern basins of Cyprus actually belong to the second stage of the Messinian salinity crisis; they can be thus ascribed to the Resedimented Lower Gypsum unit that was deposited between 5.6 and 5.5 Ma and is possibly coeval to the halite deposited in the northern Mesaoria basin. Primary, in situ evaporites of the first stage of the Messinian salinity crisis were not preserved in Cyprus basins. Conversely, shallow-water primary evaporites deposited during the third stage of the Messinian salinity crisis are well preserved; these deposits can be regarded as the equivalent of the Upper Gypsum of Sicily. Our study documents that the Messinian stratigraphy shows many similarities between the Western and Eastern Mediterranean marginal basins, implying a common and likely coeval development of the Messinian salinity crisis. This could be reflected also in intermediate and deep-water basins; we infer that the Lower Evaporites seismic unit in the deep Eastern Mediterranean basins could well be mainly composed of clastic evaporites and that its base could correspond to the Messinian erosional surface.

Aims: To assess the effect of different concentrations of gossypol (0, 2, 4, 10 and 20 mg l–1) in combination with NaCl and Na2SO4 (20 mS cm–1) on the conidial germination and viability of Fusarium oxysporum f.sp. vasinfectum (Fov). Methods and Results: A multinomial logistic model was developed to estimate the germination probability of Fov. The inhibitory effect was markedly evident at the two highest concentrations of gossypol; it varied among the isolates tested and with time, and it was attenuated by the presence of sodium salts. The inhibition was temporary as the germination probability increased after 8 h. Fluorescent staining revealed that gossypol either killed the conidia or retarded the elongation of the germ tubes. Conclusion: Fov showed the ability to overcome gossypol inhibition over time, and the inhibitory effect is reduced under saline conditions. Differential responses among Fov isolates to the presence of gossypol suggest that gossypol tolerance is genetically determined in the pathogen. Significance and Impact of the Study: This study suggests that selecting for high plant gossypol cultivars would have minimal effect on the overall Fov resistance of cotton. A new statistical model was developed to explore the statistical significance of plant–pathogen interactions.

An integrated stratigraphy (calcareous plankton biostratigraphy, magnetostratigraphy and cyclostratigraphy) is presented for the Serravallian and lower Tortonian part (Middle^Upper Miocene) of the Monte dei Corvi section located in northern Italy.The detailed biostratigraphic analysis showed that both the Discoaster kugleri acme and the first influx of Neogloboquadrina acostaensis are recorded at Monte dei Corvi; these events, which passed unobserved in previous studies, play an important role in delineating the Serravallian^Tortonian boundary.Thermal and alternating field demagnetization revealed a characteristic low-temperature component marked by dual polarities.The resultant magnetostratigraphy for the upper part of the section can be unambiguously calibrated to the GPTS ranging from C5n.2n up to C4r.2r. Unfortunately, the lower part of the section, including the Serravallian^Tortonian boundary interval, did not produce a reliable magnetostratigraphy despite the fact that some short reversed intervals and a single normal interval are recorded.Using sedimentary cycle patterns in combination with the calcareous plankton biostratigraphy the section can be correlated cyclostratigraphically in detail to the partially overlapping and previously tuned section of Monte Gibliscemi on Sicily.The Monte dei Corvi section is dated astronomically by calibrating the basic small-scale sedimentary cycles to the precession and 65‡N lat.summer insolation time series of the La93 solution following an initial tuning of larger-scale cycles to eccentricity.An almost perfect fit is found between the cycle patterns and intricate details, especially precession^obliquity interference, in the insolation target between 8.5 and 10 Ma. The tuning to precession remains robust for most intervals back to the base of the section dated at 13.4 Ma and shows that the section is continuous apart from a possible short hiatus in the Tortonian. It provides accurate astronomical ages for all sedimentary cycles, calcareous plankton events, polarity reversals and ash layers and marks a significant improvement of the recently proposed astronomical calibrations of the Monte dei Corvi section and of parallel sections in the Mediterranean.Astronomical ages for the Ancona and Respighi ashbeds are significantly older than previously reported 40 Ar/ 39 Ar biotite ages, even if the revised older age for the FCT-san dating standard of 28.02 Ma is applied. The astronomical dating of the magnetic reversals in the Monte dei Corvi section results in the completion of the astronomical polarity time scale for the last 13 Myr.The Monte dei Corvi section has recently been proposed as the stratotype section for the Serravallian^Tortonian boundary despite the moderate to