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Glucagon-like peptide-1 (GLP-1) and leptin signal recent feeding and long-term energy stores, respectively, and play complementary roles in the modulation of energy balance. Previous work using single-cell techniques in mice revealed the existence of a population of leptin receptor ( Lepr )-containing dorsomedial hypothalamus (DMH) neurons marked by the expression of GLP-1 receptor ( Glp1r ; LepR Glp1r neurons) that play important roles in the control of feeding and body weight by leptin. Here, we demonstrate the existence of a population of LepR Glp1r neurons in the DMHs of nonhuman primates (NHPs), suggesting the potential translational relevance of these neurons. Consequently, we developed a GLP-1R/LepR dual agonist and demonstrated the physiological activity of both components in vivo using leptin-deficient and Lepr- deficient murine models. We further found roles for LepR Glp1r neurons in mediating the dual agonist's efficacy on food intake and body weight loss. Ablating Lepr in Glp1r -expressing neurons (Lepr Glp1r KO mice) abrogated the suppression of food intake by the dual agonist. Furthermore, reactivation of Glp1r expression in Lepr neurons on an otherwise Glp1r -null background (Glp1r Lepr Re mice) was sufficient to permit the suppression of food intake and body weight by the dual agonist. Hence, LepR Glp1r neurons represent targets for a GLP-1R/LepR dual agonist that potently reduces food intake and body weight.

Despite extensive research, targeted delivery of substances to the brain still poses a great challenge due to the selectivity of the blood-brain barrier (BBB). Most molecules require either carrier- or receptor-mediated transport systems to reach the central nervous system (CNS). These transport systems form attractive routes for the delivery of therapeutics into the CNS, yet the number of known brain endothelium-enriched receptors allowing the transport of large molecules into the brain is scarce. Therefore, to identify novel BBB targets, we combined transcriptomic analysis of human and murine brain endothelium and performed a complex screening of BBB-enriched genes according to established selection criteria. As a result, we propose the high-affinity cationic amino acid transporter 1 (SLC7A1) as a novel candidate for transport of large molecules across the BBB. Using RNA sequencing and in situ hybridization assays, we demonstrated elevated SLC7A1 gene expression in both human and mouse brain endothelium. Moreover, we confirmed SLC7A1 protein expression in brain vasculature of both young and aged mice. To assess the potential of SLC7A1 as a transporter for larger proteins, we performed internalization and transcytosis studies using a radiolabelled or fluorophore-labelled anti-SLC7A1 antibody. Our results showed that SLC7A1 internalised a SLC7A1-specific antibody in human colorectal carcinoma (HCT116) cells. Moreover, transcytosis studies in both immortalised human brain endothelial (hCMEC/D3) cells and primary mouse brain endothelial cells clearly demonstrated that SLC7A1 effectively transported the SLC7A1-specific antibody from luminal to abluminal side. Therefore, here in this study, we present for the first time the SLC7A1 as a novel candidate for transport of larger molecules across the BBB., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Sarah Bau and Stephen T. Buckley are employees at Novo Nordisk. Catarina Chaves and Dominique Lesuisse are employees at Sanofi. Marijke De Bock is an employee at Janssen Pharmaceutica. Axel H. Mayer is an employee at AbbVie. German Leparc and Martin Lenter are employees at Boehringer Ingelheim Pharma. Tamas Letoha is the CEO of Pharmacoidea and Robert D. Bell is the Vice President and the Head of Research at Ascidian Therapeutics. All others declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier GmbH.. All rights reserved.)

This study reports size-resolved dithiothreitol (DTT)-based oxidative potential (OP: total and water-soluble) in rural kitchens using liquefied petroleum gas (LPG), firewood (FW), and mixed biomass (MB) fuels in northeastern (NE) India. In comparison to LPG, volume-normalized total OP (OP total(v) DTT ) was enhanced by a factor of ∼5 in biomass-using kitchens (74 ± 35 to 78 ± 42 nmol min -1 m -3 ); however, mass-normalized total OP (OP total(m) DTT ) was similar between LPG and FW users and higher by a factor of 2 in MB-using kitchens. The water-insoluble OP (OP wi(v, m) DTT ) fraction in OP total(v, m) DTT was greater than 50% across kitchens. Size distributions across kitchens and OP DTT categories ranged from unimodal to trimodal. OP ws(v) DTT was driven by metals as well as organics across size fractions while OP wi(v) DTT was majorly constrained by metals with an increasing importance of organics in fine particles of biomass-using kitchens. Multiple linear regression analysis revealed that Cu and Ba explained 71% of the OP total(v) DTT variability in LPG-using kitchens, while water-soluble organic carbon (WSOC) and Ba were responsible for 44% variability in FW-using kitchens. Finally, the high internal dose of OP total(v) DTT (28-31 nmol min -1 m -3 ) in biomass-using kitchens established the severity of oxidative stress on the exposed population., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

In many industrial sectors, workers are exposed to manufactured or unintentionally emitted airborne nanoparticles (NPs). To develop prevention and enhance knowledge surrounding exposure, it has become crucial to achieve a consensus on how to assess exposure to airborne NPs by inhalation in the workplace. Here, we review the literature presenting recommendations on assessing occupational exposure to NPs. The 23 distinct strategies retained were analyzed in terms of the following points: target NPs, objectives, steps, "measurement strategy" (instruments, physicochemical analysis, and data processing), "contextual information" presented, and "work activity" analysis. The robustness (consistency of information) and practical aspects (detailed methodology) of each strategy were estimated. The objectives and methodological steps varied, as did the measurement techniques. Strategies were essentially based on NPs measurement, but improvements could be made to better account for "contextual information" and "work activity". Based on this review, recommendations for an operational strategy were formulated, integrating the work activity with the measurement to provide a more complete assessment of situations leading to airborne NP exposure. These recommendations can be used with the objective of producing homogeneous exposure data for epidemiological purposes and to help improve prevention strategies., Competing Interests: The authors declare no conflict of interest., (© 2023 Occupational Safety and Health Research Institute.)

Mass-size distribution of respirable aerosol and 13 associated trace elements (TEs) were investigated in rural kitchens using liquefied petroleum gas (LPG), firewood and mixed biomass fuels across three northeastern Indian states. The averaged PM 10 (particulate matter with aerodynamic diameter ≤ 10 μm) and ΣTE concentrations were 403 and 30 μg m -3 for LPG, 2429 and 55 μg m -3 for firewood, and 1024 and 44 μg m -3 for mixed biomass-using kitchens. Mass-size distributions were tri-modal with peaks in the ultrafine (0.05-0.08 μm), accumulation (0.20-1.05 μm), and coarse (3.20-4.57 μm) modes. Respiratory deposition, estimated using the multiple path particle dosimetry model, ranged from 21 % to 58 % of the total concentration across fuel types and population age categories. Head, followed by pulmonary and tracheobronchial, was the most vulnerable deposition region, and children were the most susceptible age group. Inhalation risk assessment of TEs revealed significant non-carcinogenic as well as carcinogenic risk, especially for biomass fuel users. The potential years of life lost (PYLL) was the highest for chronic obstructive pulmonary disease (COPD: 15.9 ± 3.8 years) followed by lung cancer (10.3 ± 0.3 years) and pneumonia (10.1 ± 0.1 years), while the PYLL rate was also highest for COPD, with Cr(VI) being the major contributor. Overall, these findings reveal the significant health burden faced by the northeastern Indian population from indoor cooking using solid biomass fuels., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The relevance of dustiness methods is increasingly recognized in the preliminary exposure evaluation of workers handling nanomaterials in powder form, and should also be transposed to the assessment of environmental risk in the future . The methods currently recommended in the European standards are mainly based on determining a mass-based dustiness index [mg kg -1 ], whereas surface area is regularly put forward as a more appropriate determinant to assess the pulmonary toxicity of nanoparticles. In this study, we describe an operational methodology leading us to propose a surface metric to determine the dustiness index [m 2 kg -1 ] of nanoparticulate matter. To this end, we demonstrate the equivalence between the external specific surface area of a nanopowder and that of its aerosol with five nanomaterials produced and used on an industrial scale, and covering a range of external specific surface areas from 35 to 230 m 2 g -1 . Compared to the conventional mass-based dustiness index, the surface-based dustiness index (1) is more discriminating, covering an additional order of magnitude, and (2) has an impact on the powder ranking with potential consequences on the preventive measures to be implemented. Finally, our proposal has the potential to be included in future revisions of European standards for workplace exposure and dustiness measurement, provided that further experimental results on surface-based dustiness indices support these preliminary data.

Background: An important aspect of nanomaterial (NM) risk assessment is establishing relationships between physicochemical properties and key events governing the toxicological pathway leading to adverse outcomes. The difficulty of NM grouping can be simplified if the most toxicologically relevant dose metric is used to assess the toxicological dose-response. Here, we thoroughly investigated the relationship between acute and chronic inflammation (based on polymorphonuclear neutrophil influx (% PMN) in lung bronchoalveolar lavage) and the retained surface area in the lung. Inhalation studies were performed in rats with three classes of NMs: titanium dioxides (TiO 2 ) and carbon blacks (CB) as poorly soluble particles of low toxicity (PSLT), and multiwall carbon nanotubes (MWCNTs). We compared our results to published data from nearly 30 rigorously selected articles., Results: This analysis combined data specially generated for this work on three benchmark materials - TiO 2 P25, the CB Printex-90 and the MWCNT MWNT-7 - following subacute (4-week) inhalation with published data relating to acute (1-week) to subchronic (13-week) inhalation exposure to the classes of NMs considered. Short and long post-exposure recovery times (immediately after exposure up to more than 6 months) allowed us to examine both acute and chronic inflammation. A dose-response relationship across short-term and long-term studies was revealed linking pulmonary retained surface area dose (measured or estimated) and % PMN. This relationship takes the form of sigmoid curves, and is independent of the post-exposure time. Curve fitting equations depended on the class of NM considered, and sometimes on the duration of exposure. Based on retained surface area, long and thick MWCNTs (few hundred nm long with an aspect ratio greater than 25) had a higher inflammatory potency with 5 cm 2 /g lung sufficient to trigger an inflammatory response (at 6% PMN), whereas retained surfaces greater than 150 cm 2 /g lung were required for PSLT., Conclusions: Retained surface area is a useful metric for hazard grouping purposes. This metric would apply to both micrometric and nanometric materials, and could obviate the need for direct measurement in the lung. Indeed, it could alternatively be estimated from dosimetry models using the aerosol parameters (rigorously determined following a well-defined aerosol characterization strategy)., (© 2021. The Author(s).)

Despite recent concerns for workers' health, exposure situations to nanoparticles can occur in numerous workplaces. Understanding how exposures occur considering human work in these transformations remains a crucial issue of nanotechnologies. The objective of this article is to understand exposure situations to nanoparticles, their determinants and the resources to act on them. This understanding was achieved by specific measurement of nanoparticles aerosols, combined with an analysis of work activity (actions performed and physical strain) in a rubber industry. The presentation of real time measurements, associated with the video of work situations, during confrontation interviews becomes a means of making exposing work activities visible, to analyze and transform them from the points of view formulated by the company's stakeholders. In this way, characterized "typical exposure situations" serve to trigger discussions and open up new spaces for debate highlighting how innovation affects work and gives rise to enhanced prevention projects., Competing Interests: Declaration of competing interest The authors have no competing interests to declare., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Size-fractionated particulate mercury (PHg) measurements were performed from November 2017 to January 2018 at Terra Nova Bay (Antarctica) for the first time. Samples were collected every 10 days by a six-stage high-volume cascade impactor with size classes between 10 μm and 0.49 μm. Total PHg concentrations were maxima (87 ± 8 pg m -3 ) in November, then decreased to values ~40% lower and remained almost constant until the end of the sampling period (~30 pg m -3 ). The trimodal aerosol mass distribution reveals that from 30% to 90% of the total PHg came in the size > 1.0 μm. Hg in the two coarse fractions was probably produced by the adsorption of oxidized Hg species transported by air masses from the Antarctic plateau or produced locally by sea ice edges. PHg in accumulation mode seemed to be related to gas-particle partitioning with sea salt aerosol. Finally, average dry deposition fluxes of PHg were calculated to be 0.36 ± 0.21 ng m -2 d -1 in the accumulation mode, 47 ± 44 ng m -2 d -1 in the first coarse mode, and 37 ± 31 ng m -2 d -1 in the second coarse mode. The present work contributed to the comprehension of the Hg biogeochemical cycle, but further research studies are needed.

This study aims to characterize airborne particles emitted from a metal additive manufacturing machine and related levels of occupational exposure. To achieve this, a complete measurement methodology was deployed around a direct energy deposition machine. Different operating conditions were investigated, based on configurations of two materials and two injection nozzles. Two replicates were performed for each condition. Airborne particles emitted during repeated manufacturing cycles were measured simultaneously at the source, in the near field, in the far field and on the operator. Real-time instruments were used to characterize the machine emissions (10 nm-10 µm) associated with respirable and inhalable samplers and cascade impactors. Measurements were made during both the manufacturing process and transient operating phases. In parallel, personal exposure to hexavalent chromium was assessed. The number of particles measured for the different machining phases show that high levels of particles (> 5 × 10 5 # cm -3 , 0.3-1.3 mg m -3 inhalable particles, 0.2-6 µg m -3 Cr VI ) were emitted in the machine enclosure. The size distributions indicate that more than 90% of the particles are smaller than 250 nm. Occupational exposure to Cr VI was found to be below the LOQ of 0.098 µg m -3 for the two alloys investigated. During the machining process, near-field number and mass concentrations were ∼ 10 4 # cm -3 , and below 0.04 mg m -3 , respectively. Far-field number concentrations were also on the order of 10 4 # cm -3 throughout the whole monitoring period. The transient phase of door opening was found to result in high levels of exposure (> 10 5 # cm -3 ), which were also detected in the near-field, confirming the need to implement preventative actions. To address this issue, a collective protective measure, consisting of setting a time delay of about 8 min between the end of the manufacturing process and opening of the door, could be employed. This collective measure should also be accompanied by the wearing of personal protective equipment by the operator when an intervention in the machine enclosure is necessary.

Multi-walled carbon nanotubes (MWCNTs), which vary in length, diameter, functionalization and specific surface area, are used in diverse industrial processes. Since these nanomaterials have a high aspect ratio and are biopersistant in the lung, there is a need for a rapid identification of their potential health hazard. We assessed in Sprague-Dawley rats the pulmonary toxicity of two pristine MWCNTs (the "long and thick" NM-401 and the "short and thin" NM-403) following either intratracheal instillation or 4-week inhalation in order to gain insights into the predictability and intercomparability of the two methods. The deposited doses following inhalation were lower than the instilled doses. Both types of carbon nanotube induced pulmonary neutrophil influx using both exposure methods. This influx correlated with deposited surface area across MWCNT types and means of exposure at two different time points, 1-3 days and 28-30 days post-exposure. Increased levels of DNA damage were observed across doses and time points for both exposure methods, but no dose-response relationship was observed. Intratracheal instillation of NM-401 induced fibrosis at the highest dose while lower lung deposited doses obtained by inhalation did not induce such lung pathology. No fibrosis was observed following NM-403 exposure. When the deposited dose was taken into account, sub-acute inhalation and a single instillation of NM-401 and NM-403 produced very similar inflammation and DNA damage responses. Our data suggest that the dose-dependent inflammatory responses observed after intratracheal instillation and inhalation of MWCNTs are similar and were predicted by the deposited surface area., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Nanoparticulate powders are increasingly found in the workplace. Inhalation exposure to airborne nanoparticles (NPs) is possible throughout the life-cycle of the powders. As the toxicity of NPs has never been demonstrated, it remains essential to evaluate the risks associated with NPs in order to propose preventative measures. The first step of a risk assessment strategy consists in the identification of the 'nano' nature of a material, which suffers from a lack of an operational methodology. Here, we present a simplified and operational strategy relying on the volume specific surface area (VSSA) for nanomaterial identification, based on the recommendation stemming from the European Commission and previous work on this topic from the European Project Nanodefine. The proposed strategy was tested on a set of 15 representative industrial powders (TiO 2 , SiO 2 , CuO, and ZnO), covering a wide range of properties, and previous published data. The VSSA classification was validated via a comparison with the particle size obtained by transmission electron microscopy (TEM). It was evidenced that the VSSA is in accordance with particle size for nanomaterial powder classification. The proposed methodology involves relatively accessible methods such as thermogravimetric analysis, nitrogen adsorption and helium pycnometry and limits the detection of false negatives. Moreover, it does not imply systematic confirmation of the results with the reference particle size criterion. Our results suggest that the VSSA is a promising parameter to be used for risk assessment and should be further investigated on powder mixings to confirm its relevancy to define nanomaterial powders., Competing Interests: The authors confirm that there are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Species' movements affect their response to environmental change but movement knowledge is often highly uncertain. We now have well-established methods to integrate movement knowledge into conservation practice but still lack a framework to deal with uncertainty in movement knowledge for environmental decisions. We provide a framework that distinguishes two dimensions of species' movement that are heavily influenced by uncertainty: knowledge about movement and relevance of movement to environmental decisions. Management decisions can be informed by their position in this knowledge-relevance space. We then outline a framework to support decisions around (1) increasing understanding of the relevance of movement knowledge, (2) increasing robustness of decisions to uncertainties and (3) improving knowledge on species' movement. Our decision-support framework provides guidance for managing movement-related uncertainty in systematic conservation planning, agri-environment schemes, habitat restoration and international biodiversity policy. It caters to different resource levels (time and funding) so that species' movement knowledge can be more effectively integrated into environmental decisions.

Tuberculosis is still one of the most common respiratory diseases in Africa and worldwide and miliary tuberculosis is a regular manifestation of it. Idiopathic pulmonary hemosiderosis is a rare disease entity, presenting in children as well as adults. It is characterized by the triad of recurrent episodes of alveolar hemorrhage, presenting as hemoptysis, iron deficiency anemia and bilateral pulmonary infiltrates seen on chest X-ray. These symptoms and signs can easily be confused for other diseases i.e. miliary tuberculosis, delaying appropriate management. The etiology of idiopathic pulmonary hemosiderosis remains unclear. Diagnosis is established by lung biopsy, revealing hemosiderin laden macrophages in the alveoli. Treatment during an acute episode includes corticosteroids and/or immunosuppressive therapy, as well as supportive measures. Long-term follow-up is essential.

Because nanomaterials have been increasingly developed and used in many technology and industry sectors over the last 20 years, an increasing number of workers is likely to be exposed to airborne nanoparticles. In addition, the question of the nanomaterial characteristics that should be assessed in epidemiological studies remains open. Thus, assessing occupational exposure to airborne nanoparticles will not only rely on mass concentration and chemical composition. Rather, key parameters, such as particle size, have to be included in measurement strategies. We previously proposed a methodology to estimate the Count Median Diameter (CMD) of an aerosol based on the simultaneous size-integrated measurement of two particle concentrations, lung-deposited surface area, and number, thanks to field-portable, commercially available aerosol instruments (Nanoparticle Surface Area Monitor/Condensation Particle Counter combination). In addition to previous work, this study investigates the case of various polydisperse metal oxides, organic oil, and salt particles with CMDs ranging from 16-410 nm. Once corrected, the CMDs derived from the NSAM/CPC agree within ±20% with regard to the reference electrical mobility equivalent diameter, regardless of aerosol composition, morphology, or geometric standard deviation (GSD). Furthermore, the field-applicability of the method was tested through 6 sets of experimental data stemming from workplace measurement campaigns where different materials were produced and handled (TiO 2 , SiO 2 , Ag, Multi-Walled Carbon Nanotubes-MWCNT), covering a range of CMDs between 40 and 190 nm. All situations considered, the approach based on the combination of a NSAM and a CPC leads to a satisfying estimation of particle CMD, within ±20% compared to reference CMD.

Personal monitors based on unipolar diffusion charging (miniDiSC/DiSCmini, NanoTracer, Partector) can be used to assess the individual exposure to nanoparticles in different environments. The charge acquired by the aerosol particles is nearly proportional to the particle diameter and, by coincidence, also nearly proportional to the alveolar lung-deposited surface area (LDSA), the metric reported by all three instruments. In addition, the miniDiSC/DiSCmini and the NanoTracer report particle number concentration and mean particle size. In view of their use for personal exposure studies, the comparability of these personal monitors was assessed in two measurement campaigns. Altogether 29 different polydisperse test aerosols were generated during the two campaigns, covering a large range of particle sizes, morphologies and concentrations. The data provided by the personal monitors were compared with those obtained from reference instruments: a scanning mobility particle sizer (SMPS) for LDSA and mean particle size and a ultrafine particle counter (UCPC) for number concentration. The results indicated that the LDSA concentrations and the mean particle sizes provided by all investigated instruments in this study were in the order of ±30% of the reference value obtained from the SMPS when the particle sizes of the test aerosols generated were within 20-400nm and the instruments were properly calibrated. Particle size, morphology and concentration did not have a major effect within the aforementioned limits. The comparability of the number concentrations was found to be slightly worse and in the range of ±50% of the reference value obtained from the UCPC. In addition, a minor effect of the particle morphology on the number concentration measurements was observed. The presence of particles >400nm can drastically bias the measurement results of all instruments and all metrics determined., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Measurement of worker exposure to a thoracic health-related aerosol fraction is necessary in a number of occupational situations. This is the case of workplaces with atmospheres polluted by fibrous particles, such as cotton dust or asbestos, and by particles inducing irritation or bronchoconstriction such as acid mists or flour dust. Three personal and two static thoracic aerosol samplers were tested under laboratory conditions. Sampling efficiency with respect to particle aerodynamic diameter was measured in a horizontal low wind tunnel and in a vertical calm air chamber. Sampling performance was evaluated against conventional thoracic penetration. Three of the tested samplers performed well, when sampling the thoracic aerosol at nominal flow rate and two others performed well at optimized flow rate. The limit of flow rate optimization was found when using cyclone samplers., (© The Author 2017. Published by Oxford University Press on behalf of the British Occupational Hygiene Society.)

Background : Hepatitis C (HCV) direct acting antiviral agents (DAAs) are safe, effective, and tolerable. Most contraindications to interferon-based treatment are no long applicable. The aims of this study were to understand the predictors of approval to drug accessibility. Methods : We studied all consecutive patients with HCV prescribed DAAs between October 2014 and July 2015. Data on demographic, socio-economic status, comorbidities, baseline laboratory values, and assessment of liver disease severity, insurance, and specialty pharmacy type were collected. Multivariate analyses were performed to identify predictors of prescription approval. Results : In total, 410 patients were prescribed DAAs between October 2014 and July 2015. Of those, 332 (81%) patients were insurance approved for therapy. Of the 332 patients accepted, 251 were accepted after the first prescription attempt, and 38 were accepted after the second and third attempts. The number of attempts for the other 43 approved patients was unknown. Older age (p = 0.001), employment (p = 0.001), lack of comorbidities (p = 0.02), liver transplantation (p = 0.018), and advanced liver disease (p = 0.001) were more likely associated with obtaining approval. Household income was not associated with insurance approval. In the multivariate analysis, Medicare insurance (odds ratio [OR]) 2.67, 95% confidence interval [CI] 0.96-7.20), lack of nonliver comorbidities (OR 2.72, 95% CI 1.35-5.43), and the presence of advanced liver disease (OR 1.82, 95% CI 1.04-3.24) independently predicted drug approval. Conclusion : Despite the availability of DAAs for HCV, barriers from insurance carriers continue to impair widespread use. Patients with advanced liver disease, Medicare, and without comorbidities are most likely to be insurance approved for DAAs.

Background and Aims: All-oral interferon-free antivirals are highly effective in treating recurrent hepatitis C (HCV) infection in liver transplant (LT) recipients. The aim of the study was to assess immunosuppression needs after achieving a sustained viral response (SVR)., Methods: We compared immunosuppression needs before and after achieving a SVR in adult LT recipients treated for recurrent HCV infection with all-oral direct acting agents., Results: We identified 52 liver LT treated recipients who achieved a SVR. The median (25th and 75th percentile interquartile range [IQR]) age was 62 years (57.75, 65). Most recipients received tacrolimus (TAC) for their immunosuppressant regimen. After achieving SVR, there was no statistically significant difference in daily dose of TAC unadjusted per weight (p > 0.05). However, there was a statistically significant decrease in daily dose of TAC adjusted per weight, serum levels of TAC, and the product of glomerular filtration rate and TAC. No statistically significant differences in cyclosporine unadjusted/adjusted per weight daily dose or serum levels were noted., Conclusions: Immunosuppression needs were increased for those patients treated with TAC but not cyclosporine. LT recipients prescribed TAC require close monitoring after treatment completion to avoid potential risk of acute rejection.

Substantial advances have been made in our understanding of the movement of species, including processes such as dispersal and migration. This knowledge has the potential to improve decisions about biodiversity policy and management, but it can be difficult for decision makers to readily access and integrate the growing body of movement science. This is, in part, due to a lack of synthesis of information that is sufficiently contextualized for a policy audience. Here, we identify key species movement concepts, including mechanisms, types, and moderators of movement, and review their relevance to (1) national biodiversity policies and strategies, (2) reserve planning and management, (3) threatened species protection and recovery, (4) impact and risk assessments, and (5) the prioritization of restoration actions. Based on the review, and considering recent developments in movement ecology, we provide a new framework that draws links between aspects of movement knowledge that are likely the most relevant to each biodiversity policy category. Our framework also shows that there is substantial opportunity for collaboration between researchers and government decision makers in the use of movement science to promote positive biodiversity outcomes.

Background: Fibrosing cholestatic hepatitis (FCH) is an uncommon but potentially fatal complication of recurrent hepatitis C (HCV) in liver transplant recipients., Methods: We matched the treatment outcomes of 10 liver transplant recipients who developed FCH with those of 10 recipients with recurrent HCV without FCH treated with sofosbuvir and ribavirin., Results: Baseline mean alanine transaminase, aspartate transaminase, alkaline phosphatase, and total bilirubin were 186 U/L, 197 U/L, 243 U/L, and 6.7 mg/dL, respectively, in the FCH recipients and 82 U/L, 60 U/L, 110 U/L, and 0.99 mg/dL, respectively, in non-FCH recipients. The sustained viral response in FCH and non-FCH recipients was 40% and 80%, respectively. One-yr patient and graft survival rates were 90% and 80%, respectively, in FCH recipients, and 100% in non-FCH recipients. Seven FCH and six non-FCH recipients were treated for anemia with blood transfusion and/or erythropoietin growth factors., Conclusion: Our results suggest that the use of sofosbuvir and ribavirin is effective and tolerable in liver transplant recipients treated for recurrent FCH. There is a trend of lower sustained viral response, patient survival, and graft survival in the FCH recipients., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

In addition to chemical composition, particle concentration and size are among the main parameters used to characterize exposure to airborne ultrafine or nanoparticles. To assess occupational inhalation exposure, real-time instruments are recommended in recent strategies published. Among portable devices for personal exposure assessment in the workplace, DiSCmini (Matter Aerosol AG, Switzerland) has been identified as a potential candidate with its capacity to measure the airborne nanoparticle concentration and average particle size with good time-resolution. Monodisperse and polydisperse test nanoaerosols of varying compositions and morphologies were produced in the laboratory using the CAIMAN facility. These aerosols covered a range of particle sizes between 15 and 400 nm and number concentrations from 700 to 840,000 cm(-3). The aerosols were used to investigate the behavior of DiSCmini, comparing experimental data to reference data. In spite of a slight tendency to underestimate particle size, all particle diameters, number concentrations and surface area concentrations measured were in the same order of magnitude as reference data. Furthermore, no significant effect due to particle composition or morphology was noted.

Background: There has been increasing interest in using protease inhibitors with pegylated interferon and ribavirin to treat recurrent hepatitis C (HCV) disease in liver transplant recipients., Methods: We retrospectively evaluated the safety and efficacy in liver transplant recipients treated for recurrent hepatitis C genotype 1 with the combination of peginterferon, ribavirin and boceprevir., Results: Twenty liver transplant recipients were treated for recurrent hepatitis C. Baseline alanine aminotransferase, total bilirubin and HCV RNA values (± SD) were 67.5 (±50.9) mg/dl, 1.78 (±1.99) U/L, and 16 955 510 (±21 620 675) IU/ml. Anaemia was a common adverse event requiring epoetin in 16 of 20 recipients and ribavirin dose reductions in 17 of 20 recipients. One-third of recipients required a blood transfusion. Filgrastim was used in 11 of 20 patients (55%) and eltrombopag in two of 20 recipients (10%) over the course of treatment. Serum creatinine level increased significantly from a baseline value of 1.33 mg/dl to 1.59 mg/dl at week 20 of boceprevir (P < 0.005). The overall sustained viral response (SVR) was 50%. Of the 14 patients who had a viral load less than 1000 IU/ml at week 4 of boceprevir, the SVR was 71%. The SVR was 83% of the 11 patients who had undetectable viral levels at week 4 of boceprevir., Conclusions: Antiviral therapy utilizing boceprevir in liver transplant recipients requires close monitoring. Anaemia and neutropenia were common requiring growth factors in most recipients. On-treatment viral responses appear promising but long-term data are needed., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Despite improved virologic response with the addition of direct acting agents to peginterferon and ribavirin treatment in chronic hepatitis C virus (HCV) genotype 1 infection, a subset of patients experience viral breakthrough while on therapy. Defining viral breakthrough and patient characteristics is important for ongoing and future HCV treatment., Methods: Thirty-four patients treated with telaprevir between June 2011 and July 2012 were retrospectively evaluated for presence of viral breakthrough. Baseline patient characteristics, time to viral breakthrough, and HCV resistance patterns were determined., Results: Viral breakthrough was seen in 26.5% of patients treated. Eight of 9 patients experienced breakthrough in the peginterferon and ribavirin-only phase of treatment with mean (±SD) time to breakthrough of 21.3 (±6.4) weeks. Viral breakthrough was more frequently seen in patients with genotype 1a, advanced liver fibrosis, and prior null treatment response. A majority of patients had presence of resistant mutations upon testing., Conclusions: A significant proportion of patients experience viral breakthrough after completion of the direct acting agent portion of triple therapy. More frequent virologic assessments during the peginterferon and ribavirin treatment phase may be necessary to reduce cost and adverse effects of treatment.

In the frame of assessing exposure to nanostructured particles, the aim of this work is to study the performance of a new device devoted to the real-time measurement of nanostructured aerosol: the meDiSC (Diffusion Size Classifier, Matter Engineering, Switzerland). This instrument is based on unipolar diffusion charging of particles which are then collected successively in diffusion and filtration stages. From currents measured in these stages, the instrument is capable of determining aerosol mean size and number concentration. These data were compared to reference measurements regarding monodisperse aerosols in a range from 20 to 700 nm; the relative biases were found unsatisfying. This led us to investigate the principle of the instrument. Consequently, the charging law of the diffusion charger was experimentally established, as well as the calibration curve allowing the determination of the mean size of the particles. The latter analysis was completed by a model based on diffusion theory. Our results indicate the possibility to improve the range of size measurement up to 350 nm. Measured particle size and number concentration were also used to calculate geometric surface-area concentration; the experimental data were compared to a reference calculated surface-area concentration. The results demonstrate the possibility to evaluate this parameter within acceptable uncertainty. In a second step, the meDiSC was challenged with polydisperse aerosols. It was observed that meDiSC overestimates particle size by a factor 1.7, while particle number concentrations are found within ±40% of the reference. The model applied to polydisperse aerosols indicates that polydispersity little influences particle size up to 300 nm while geometric standard deviation remains below 1.7., (This journal is © The Royal Society of Chemistry 2012)

Sequence capture methods for targeted next generation sequencing promise to massively reduce cost of genomics projects compared to untargeted sequencing. However, evaluated capture methods specifically dedicated to biologically relevant genomic regions are rare. Whole exome capture has been shown to be a powerful tool to discover the genetic origin of disease and provides a reduction in target size and thus calculative sequencing capacity of >90-fold compared to untargeted whole genome sequencing. For further cost reduction, a valuable complementing approach is the analysis of smaller, relevant gene subsets but involving large cohorts of samples. However, effective adjustment of target sizes and sample numbers is hampered by the limited scalability of enrichment systems. We report a highly scalable and automated method to capture a 480 Kb exome subset of 115 cancer-related genes using microfluidic DNA arrays. The arrays are adaptable from 125 Kb to 1 Mb target size and/or one to eight samples without barcoding strategies, representing a further 26 - 270-fold reduction of calculative sequencing capacity compared to whole exome sequencing. Illumina GAII analysis of a HapMap genome enriched for this exome subset revealed a completeness of >96%. Uniformity was such that >68% of exons had at least half the median depth of coverage. An analysis of reference SNPs revealed a sensitivity of up to 93% and a specificity of 98.2% or higher., (Copyright 2010 Elsevier Inc. All rights reserved.)

We report a flexible method for selective capture of sequence fragments from complex, eukaryotic genome libraries for next-generation sequencing based on hybridization to DNA microarrays. Using microfluidic array architecture and integrated hardware, the process is amenable to complete automation and does not introduce amplification steps into the standard library preparation workflow, thereby avoiding bias of sequence distribution and fragment lengths. We captured a discontiguous human genomic target region of 185 kb using a tiling design with 50mer probes. Analysis by high-throughput sequencing using an Illumina/Solexa 1G Genome Analyzer revealed 2150-fold enrichment with mean per base coverage between 4.6 and 107.5-fold for the individual target regions. This method represents a flexible and cost-effective approach for large-scale resequencing of complex genomes.

Objective: The purpose of this study was to evaluate the accuracy of 3-dimensional (3D) sonography in assessing fetal anatomy and to determine the intraobserver reproducibility and the effect of examiner experience., Methods: Three-dimensional volumes of the head, face, thorax, and abdomen were obtained for 40 fetuses. The volume data sets obtained were explored offline with multiplanar navigation and tomographic ultrasound imaging on a personal computer. Each case was examined twice by the same observer at least 3 months apart. The percentage for identification of fetal anatomic structures, 2-dimensional (2D) and 3D measurements, and the time spent on 2D and 3D examinations were calculated and compared., Results: Ninety-two percent of fetal anatomic structures were identified with multiplanar navigation and tomographic ultrasound imaging. The genitals, the entry of the vena cava, and the ears were visualized in less than 70% of cases. Tomographic ultrasound imaging allowed viewing of 14 structures not seen in the multiplanar study. Intraobserver agreement for anatomic examinations was good (kappa = 0.78). Intraobserver agreement for fetal measurements showed differences between both examinations that approached 0 and intraclass correlation indices close to 0.9. The mean 2D scanning time +/- SD was 10.11 +/- 2.5 minutes, and the acquisition time for the 3D volumes was 1.54 +/- 0.35 minutes. The total time for the second 3D study was 7.23 +/- 1.17 minutes, significantly shorter than the 9.96 +/- 1.53 minutes spent on the first study (P < .001)., Conclusions: Three-dimensional volumes are highly effective for complete fetal anatomic surveys. They show excellent intraobserver reproducibility and take less time to study as the examiner's experience increases.

Cell-autonomous immunity is widespread in plant-fungus interactions and terminates fungal pathogenesis either at the cell surface or after pathogen entry. Although post-invasive resistance responses typically coincide with a self-contained cell death of plant cells undergoing attack by parasites, these cells survive pre-invasive defence. Mutational analysis in Arabidopsis identified PEN1 syntaxin as one component of two pre-invasive resistance pathways against ascomycete powdery mildew fungi. Here we show that plasma-membrane-resident PEN1 promiscuously forms SDS-resistant soluble N-ethylmaleimide sensitive factor attachment protein receptor (SNARE) complexes together with the SNAP33 adaptor and a subset of vesicle-associated membrane proteins (VAMPs). PEN1-dependent disease resistance acts in vivo mainly through two functionally redundant VAMP72 subfamily members, VAMP721 and VAMP722. Unexpectedly, the same two VAMP proteins also operate redundantly in a default secretory pathway, suggesting dual functions in separate biological processes owing to evolutionary co-option of the default pathway for plant immunity. The disease resistance function of the secretory PEN1-SNAP33-VAMP721/722 complex and the pathogen-induced subcellular dynamics of its components are mechanistically reminiscent of immunological synapse formation in vertebrates, enabling execution of immune responses through focal secretion.

Objective: To evaluate whether endometrial parameters by three-dimensional ultrasonography and power Doppler angiography (3D US-PDA) can predict in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) outcome., Design: Prospective clinical study., Setting: Assisted reproduction unit in a referral hospital., Patient(s): Eighty women who underwent IVF cycles., Intervention(s): Endometrial 3D US-PDA evaluated by VOCAL software (plane C and 9 degrees of rotational steps)., Main Outcome Measure(s): Endometrial pattern, endometrial thickness (ET), endometrial volume (EV), and PDA indexes of vascularization index (VI), flow index (FI), and vascularization flow index (VFI) were measured on the day of human chorionic gonadotropin (hCG) administration. These measurements were related to IVF/ICSI and embryo transfer outcome., Result(s): In the pregnant group, EV, VI, FI, and FVI but not triple-line pattern and ET were statistically significantly higher. The area under receiver operating characteristic (ROC) curve was statistically significant for EV (0.746), VI (0.724), FI (0.828), and VFI (0.800) when no grade 1 embryos or only one were transferred (43 cycles, 14 pregnancies) but not when two or three grade 1 embryos were transferred. Moreover, these parameters were statistically significant in predicting a normal pregnancy outcome (no early pregnancy loss) but were not related to multiple pregnancies., Conclusion(s): In IVF/ICSI cycles, 3D US-PDA is useful for evaluating endometrial receptivity. Endometrial volume and 3D power Doppler indexes are statistically significant in predicting the cycle outcome when one grade 1 or no grade 1 embryos are transferred, which could be helpful data in a single-embryo transfer policy.

Objective: The purpose of this study was to assess whether endometrial volume (EV) and 3-dimensional (3D) power Doppler indices can discriminate between hyperplasia and endometrial carcinoma and can predict extension of the endometrial carcinoma., Methods: Eighty-four women with uterine bleeding and a histopathologic diagnosis of endometrial hyperplasia (n = 29) or carcinoma (n = 55) were preoperatively examined by transvaginal 3D sonography and power Doppler angiography. Endometrial thickness (ET), EV, the vascularization index (VI), the flow index (FI), the vascularization-flow index (VFI), and the intratumoral resistive index (RI) were measured. A histopathologic diagnosis was made after endometrial biopsy was performed by hysteroscopy or curettage., Results: The EV and 3D power Doppler indices (VI, FI, and VFI) were significantly higher in endometrial carcinoma than endometrial hyperplasia, whereas the intratumoral RI was significantly lower (P < .05). A VFI of 2.07 was the best cutoff for predicting endometrial carcinoma, with sensitivity of 76.5% and specificity of 80.8%. No significant differences were noticed for ET. The endometrial VI was significantly higher when the tumor stage was greater than I. All the 3D power Doppler indices were significantly higher when the carcinoma infiltrated more than 50% of the myometrium. The intratumoral RI was significantly lower in cases with a high histologic grade, myometrial infiltration of more than 50%, and lymph node metastases., Conclusions: The VI, 3D power Doppler indices, and the intratumoral RI are more useful than ET for differentiating between hyperplasia and endometrial carcinoma. Intratumoral blood flow evaluated by pulsed Doppler sonography and 3D power Doppler angiography can predict the spread of endometrial carcinoma.

Objective: The objective was to evaluate whether three-dimensional ultrasonography (3D-US) and power Doppler angiography (PDA) measurements can predict ovarian response and/or are associated with IVF/ICSI outcome., Study Design: A prospective clinical study in 65 women undergoing IVF cycles. Ovarian volume (OV), number of antral follicles > or =2mm (NAF) and PDA indices: vascularisation index (VI), flow index (FI), and vascularisation flow index (VFI) were evaluated by 3D-US and PDA on the day of pituitary suppression control. These measurements, age and BMI were correlated with the number of follicles>10mm on the hCG day and the number of oocytes retrieved., Results: Ovarian volume, NAF, VI, FI and FVI correlate significantly (P<0.01) with the number of follicles and oocytes recovered. Ovarian volume and the number of antral follicles predicted significantly the number of follicles (R=0.67; adjusted R(2)=0.43) and oocytes retrieved (R=0.63; adjusted R(2)=0.37). The oestradiol peak and the number of follicles, oocytes and Grade 1 embryos transferred were higher in the pregnant group., Conclusions: Three-dimensional ultrasound and PDA make it easier to evaluate all the sonographic parameters implied in ovarian response. Ovarian volume and the number of antral follicles are the only independent predictors of the number of follicles developed and oocytes retrieved.

Objective: The aim of this prospective study was to investigate whether ovarian blood flow is related to embryological parameters and whether it could be a predictor of outcomes of IVF/ICSI., Methods: Eighty infertile women underwent ovarian stimulation with gonadotrophins after a long protocol with GnRH agonists. The ovarian volume (OV), number of follicles (NF) and follicular volume (FV) of all follicles >10 mm and vascularization index (VI), flow index (FI) and vascularization-flow index (VFI) were obtained by three-dimensional (3D) ultrasonography and power Doppler angiography (PDA) on the day of HCG administration. These parameters were tested for their relation with IVF laboratory parameters., Results: The OV, FV, VI, FI and VFI were significantly greater in the pregnant group. The NF and FV were the only independent predictors of the number of oocytes retrieved, mature and fertilized, and the number of embryos developed and their cumulative embryo score. Nevertheless, the number of grade 1 embryos depends on the NF and the VI. The ovarian FI and the number of transferred grade 1 embryos can predict gestation in 76% of IVF patients. A low FI and non-grade 1 embryo transferred are also associated with an increased pregnancy loss., Conclusion: 3D ultrasonography and PDA allow for an easier ovarian assessment in IVF cycles. The predictive value of IVF outcome suggests a high clinical usefulness of this new technique.

Objectives: To assess intraobserver reproducibility of the endometrial volume (EV) and 3D power Doppler indices (vascularization index, VI; flow index, FI; and vascularization flow index, VFI) of the endometrium and subendometrial area using three-dimensional power Doppler angiography (3D-PDA)., Methods: Twenty-five women on the hCG day after controlled ovarian stimulation and 15 patients presenting with uterine bleeding and suspicious endometrial thickening (10 endometrial cancers and 5 endometrial hyperplasias) were scanned. Eighty volume data sets were analyzed using the VOCAL imaging program. EV and VI, FI and VFI of the endometrium and subendometrium (5 mm shell) were manually calculated in the longitudinal and coronal planes with 15 degrees and 9 degrees rotation steps. Intraclass correlation coefficient (ICC) and 95% confidence intervals were used to assess reliability. RESULTS.: EV measurements were highly reproducible (ICC > or = 0.97) without significant differences between planes and rotation steps. Endometrial and subendometrial VI, FI, and VFI presented ICCs above 0.90 with the exception of the subendometrial FI (ICC > or = 0.80). There were no significant differences according to measurement plane and rotation step except for subendometrial VFI. Nevertheless, 3D power Doppler indices calculated in the coronal plane and 9 degrees rotation step obtained the highest ICC. ICCs for 3D-PDA indices from the tumoral endometria were significantly higher than those calculated from the stimulated endometria., Conclusions: Endometrial volume and endometrial and subendometrial 3D power Doppler indices have an acceptable reproducibility, significantly higher in tumoral endometria. The reliability of measurements does not seem to be significantly influenced by the rotation plane and degrees of rotation. These results support that 3D-PDA and VOCAL are reliable methods to evaluate the physiological and pathological changes of the endometrium.

Aim: To describe the evolution of placental vascularization during a normal course of gestation and the blood flow 3D power Doppler indices obtained by "placental vascular biopsy.", Methods: A prospective study was carried out on 99 normal singleton pregnancies from 14 to 40 weeks. Placental vascularization was evaluated by 3D power Doppler ("placental biopsy"). The spherical volume acquired was analyzed using the VOCAL imaging program (Virtual Organ Computer-aided AnaLysis). Three vascular indices, vascularization index (VI), flow index (FI), and vascularization-flow index (VFI), were calculated. Equations and regression coefficients for placental volume and vascular indices (VI, FI, VFI) of the placental biopsy were calculated according to gestational age. Relationships between 3D placental flow indices and fetal growth parameters: biparietal diameter, head circumference, abdominal circumference, femur length, estimated fetal body weight, maximum systolic velocity (US), and resistance index in the umbilical artery (URI) were evaluated by calculating their correlation coefficients., Results: All 3D Doppler indices had a significant relationship with gestational age. The most significant relationship was observed for FI, and the least significant for VI (r=0.58, r=0.29, respectively; P<0.01 for both). The FI increased linearly with gestation, whereas the VI showed a dispersion of values with a plateau from the 30th week onwards and a decrease from 37th week to the end of pregnancy. The VFI behaved as a combination of both VI and FI indices from which it was derived. All 3D Doppler indices were significantly related to fetal biometric parameters, except VI and fetal weight. A significant correlation was observed between 3D Doppler indices and maximum systolic velocity and URI., Conclusions: 3D power Doppler technique of placental vascular "biopsy" is an appropriate tool for routine evaluation of the human placental vascular tree during gestation. 3D Doppler indices change as pregnancy progresses and are significantly related with fetal biometry and umbilical artery Doppler velocimetry.

Objectives: The purpose of this study was to assess intraobserver and interobserver reproducibility of the parameters of ovarian response and oocyte ability, studied by 3-dimensional ultrasonography and power Doppler angiography (PDA), and the possible influence of the ovarian functional stage., Methods: Twenty-nine women were included in an in vitro fertilization program. Fourteen women were evaluated after pituitary suppression (basal group), and 15 were scanned on the human chorionic gonadotropin administration day, after gonadotropin ovarian stimulation (stimulated group). A first observer acquired 2 volumes for each ovary. Another observer performed a second analysis of the volumes acquired by the first observer. We analyzed ovarian volume, follicle number in the basal group, vascularization index, flow index, and vascularization-flow index. The volumes were processed by the Virtual Organ Computer-Aided Analysis imaging program using plane A and 15 degrees rotational steps., Results: Ovarian volume showed excellent intraobserver and interobserver agreement, with an intraclass correlation coefficient (Intra-CC) and an interclass correlation coefficient (Inter-CC) close to the unit. The Intra-CC and Inter-CC about the number of follicles were 0.964 and 0.978, respectively. Vascularity indices showed an Intra-CC greater than 0.90. The vascularization index and the vascularization-flow index showed higher interobserver than intraobserver reproducibility (Inter-CC > 0.96 for both). The flow index Inter-CC was 0.898. The reproducibility differences between the basal and stimulated ovary measurements were not significant., Conclusions: There is an excellent intraobserver and interobserver reproducibility of the ovarian volume, follicle counts, and 3-dimensional PDA indices. The ovarian functional stage has no influence on the reliability. Three-dimensional ultrasonography and PDA improve the study of ovarian parameters, and their reliability impels a change in the current clinical routine of performing and interpreting ultrasonography.

Objective: The purpose of this study was to evaluate interobserver reproducibility of endometrial volume and vascular indices of the endometrium and subendometrial area estimated by 3-dimensional power Doppler angiography (3D-PDA) using the Virtual Organ Computer-Aided Analysis program, determining the influence of the endometrial growth etiology on measurements., Methods: Forty women underwent 3D-PDA ultrasonography. Group A comprised 25 women scanned on the day after controlled ovarian stimulation with human chorionic gonadotropin. Group B comprised 15 patients who had uterine bleeding and questionable endometrial thickening. (Histologic evaluation revealed 10 endometrial cancers and 5 endometrial hyperplasias.) A single observer examined all patients and acquired all volume data sets. Forty volume data sets were then analyzed with the Virtual Organ Computer-Aided Analysis program by 2 different observers. Endometrial volume and vascularity indices (vascularization index [VI], flow index [FI], and vascularization flow index [VFI]) of the endometrium and subendometrium were manually calculated in the coronal plane with a 9 degrees rotation step. An intraclass correlation coefficient (ICC) was used to assess interobserver reliability., Results: Endometrial volume was more reproducible in group A (ICC = 0.98) than in group B (ICC = 0.58) (P < .05). Endometrial and subendometrial VI, FI, and VFI also presented good reproducibility with ICC greater than 0.84. The ICC was not statistically different for endometrial and subendometrial VI, FI, and VFI according to patient group, although subendometrial VFI was less reproducible in group B (ICC = 0.53) than in group A (ICC = 0.88)., Conclusions: Endometrial volume and endometrial and subendometrial 3D power Doppler indices have acceptable reproducibility. The interobserver reproducibility in tumoral endometrium was more similar than in stimulated endometrium. Our results indicate that 3D-PDA is a reliable method to evaluate physiologic and pathologic endometrial changes.

Aims: To assess the reproducibility of 3D power Doppler study of placental vascularization in order to establish its methodological bases for its further application in normal and pathological pregnancies., Methods: A prospective study was carried on 30 normal singleton pregnancies from 14 to 40 weeks. To evaluate placental vascularization 3D power Doppler was applied to obtain a "placental biopsy". The spherical volume acquired was analyzed using the VOCAL imaging program. Two consecutive measurements were taken from each patient by a single observer, obtaining a total of 60 datasets. Placental volume (PV), Mean Gray (MG), Vascularization Index (VI), Flow Index (FI) and Vascularization Flow Index (VFI) were calculated. Intra-class correlation coefficient (ICC) and intra-observer agreement was evaluated., Results: PV and MG presented an ICC of 0.98 and 0.94 respectively, with differences approaching zero. All 3D power Doppler vascular indices (VI, FI and VFI) showed a correlation greater than 0.85, with a better intra-observer agreement for the flow indices (FI and VFI)., Conclusions: Placental vascular biopsy through 3D power Doppler is a new and simple tool to routinely study placental vascularization in human pregnancy. Our results provide the validation of the technique demonstrating a good reproducibility of the 3D power Doppler parameters when applied to the study of the placental vascular tree in normal pregnancies.

Failure of pathogenic fungi to breach the plant cell wall constitutes a major component of immunity of non-host plant species--species outside the pathogen host range--and accounts for a proportion of aborted infection attempts on 'susceptible' host plants (basal resistance). Neither form of penetration resistance is understood at the molecular level. We developed a screen for penetration (pen) mutants of Arabidopsis, which are disabled in non-host penetration resistance against barley powdery mildew, Blumeria graminis f. sp. hordei, and we isolated the PEN1 gene. We also isolated barley ROR2 (ref. 2), which is required for basal penetration resistance against B. g. hordei. The genes encode functionally homologous syntaxins, demonstrating a mechanistic link between non-host resistance and basal penetration resistance in monocotyledons and dicotyledons. We show that resistance in barley requires a SNAP-25 (synaptosome-associated protein, molecular mass 25 kDa) homologue capable of forming a binary SNAP receptor (SNARE) complex with ROR2. Genetic control of vesicle behaviour at penetration sites, and plasma membrane location of PEN1/ROR2, is consistent with a proposed involvement of SNARE-complex-mediated exocytosis and/or homotypic vesicle fusion events in resistance. Functions associated with SNARE-dependent penetration resistance are dispensable for immunity mediated by race-specific resistance (R) genes, highlighting fundamental differences between these two resistance forms.

The aims of this study were to determine, firstly, the relationship between FIGO stage and various tumor parameters determined by magnetic resonance imaging (MRI), and, secondly, whether any of these parameters were predictors of lymph node metastases as determined by fluorine-18 fluorodeoxyglucose positron emission tomography (FDG PET) in cervical cancer patients referred for radiotherapy. In 70 consecutive patients, both PET and MRI visualized all primary tumors except for one previously removed by cone biopsy. While clinical diameter and MRI-derived diameter showed a significant relationship between these two measurements (r = 0.70; P < 0.001) there was a large variability in MRI diameter for each FIGO stage and wide overlap. The average volume of primary cervical tumor on MRI was 60 cc (5-256). In FIGO stages, I, II, III and IV, uterine body involvement was present in 58%, 73%, 88%, and 100% of 19, 30, 16, and 5 patients, respectively (Ptrend= 0.015). Node positivity on FDG PET was present in 11% of patients without uterine body extension, but increased to 75% in those with uterine involvement. Average tumor volume in node-negative patients was 49 cc (5-186). Average tumor volume in node-positive patients was 69 cc (8-256). There was a significant association between nodal involvement and both FIGO stage (P = 0.018) and uterine body involvement (P < 0.001), but tumor volume and longitudinal MRI diameter were not statistically significant in unifactor predictors of nodal involvement. In multivariate analysis only uterine body extension, however, was independently related to the risk of nodal involvement. In conclusion, MRI provides noninvasive tumor size evaluation and can also demonstrate invasion of the uterine body that appears to be associated with an increased risk of nodal metastasis. This may provide clinically important prognostic information not available from current FIGO staging.

Background: The purpose of this paper is to review and report our experience with colorectal stenting in the management of malignant large bowel obstruction., Methods: Twelve consecutive patients with malignant left-sided large bowel obstruction between June 1998 and January 2001 underwent insertion of self-expanding metallic stents. One patient required two stents. Eight stents were inserted under fluoroscopic guidance, and five were inserted with combined fluoroscopic and endoscopic guidance. Patients were followed up until death, stent removal or the time of publication., Results: Thirteen stents were inserted. Eleven patients with acute large bowel obstruction had relief of obstruction with stenting, and one of these patients required a second stent because relief had been incomplete. One patient was stented in order to subsequently close a problematic stoma. Technical success was 92.9% and clinical success was 100%. Three patients proceeded to surgical resection. In nine patients, the stent was left as the definitive procedure. Of these, six patients have died within 4 months. Complications included one case of migration, one case of reobstruction, one intestinal haematoma and one case of cheesewiring., Conclusions: Colorectal stenting is an important treatment modality for malignant large bowel obstruction. It may be definitive treatment alone, or a bridge to elective surgical resection.

Objective: To evaluate arterial and venous intraovarian blood flow in follicle stimulating hormone-stimulated cycles., Subjects and Methods: This was a prospective study of 76 follicle stimulating hormone-stimulated cycles carried out in 39 infertile patients who were included in a timed intercourse or intrauterine insemination program in a referral center for assisted reproduction. Transvaginal color and pulsed Doppler measurements of the follicular and luteal phase resistance index, pulsatility index, peak systolic velocity and maximum venous velocity were made and serum progesterone levels during the mid-luteal phase were recorded. Velocimetric parameters were established and then used to classify ovarian function as having a normal ovulatory cycle, or a cycle in which there was either luteal phase deficiency or a luteinized unruptured follicle., Results: In 52 normal ovulatory cycles, the luteal phase peak systolic and maximum venous velocities were significantly higher and resistance and pulsatility indices were significantly lower than those found in the follicular phase. In 15 women with luteal phase deficiency we did not find any differences in arterial velocimetric parameters when compared with normal ovulatory cycles. However, luteal phase maximum venous velocities were lower in the luteal phase deficiency cycles and there was a significant correlation between luteal phase maximum venous velocity and serum progesterone levels (r = 0.36). Luteinized unruptured follicle cycles (n = 9) did not show significant changes during the ovarian cycle and no 'luteal conversion' of the Doppler signal was identified., Conclusions: Follicle stimulating hormone-stimulated cycles in infertile patients can have a high percentage of abnormal functional responses that can be diagnosed only by sonographic assessment, Doppler and the appropriate hormonal follow-up. Arterial and venous intraovarian blood flow remain unaltered during luteinized unruptured follicle cycles and serum progesterone levels correlated with luteal phase maximum venous velocity, which makes Doppler a potentially useful non-invasive test to assess ovulation and luteal function.

Objectives: The aim of this study was to evaluate and compare the use of B-mode transvaginal, utero-ovarian Doppler and intratumoural colour Doppler imaging in the diagnosis of ovarian tumours in order to establish whether they are organic or functional, and malignant or benign in nature prior to surgery., Study Design: A series of 213 women between the ages of 15 and 87 presenting ovarian tumours were examined with pulsed and colour Doppler and the functional state of the ovary at the time of the exploration was also established. Size and morphology were evaluated through a new scoring system called sonographic index. This score is calculated as the sum of the points corresponding to tumoural volume (< or = 8 ml = 1 point; > 8 ml and < or = 100 ml = 2 points; > 100 ml = 3 points) and the points corresponding to morphological characteristics (negative echogenicity = 1 point; mixed echogenicity = 2 points; multilocular morphology = 3 points; complex morphology = 4 points; positive echogenicity = 5 points) varying between 2 and 8 points. The flow velocity waveform (FVW) of the utero-ovarian and intratumoural arteries were obtained and analyzed after assessing their resistance indices. Other parameters studied were the presence and location of the colour signal in the intratumoural arteries., Results: Of the total, 84 of the tumours were found to be functional and had disappeared by the time later examinations took place. Surgery was performed in 129 of the patients, and posterior histopathological studies permitted classifying the tumours as benign in 107 cases and malignant in 22. The sonographic index was significantly higher in the organic (4.9 +/- 1.5) and malignant (6.9 +/- 1.0) tumours than in the functional (3.6 +/- 1.5) and the benign (4.1 +/- 1.2) tumours. The utero-ovarian and intratumoural blood flow RIs in the malignant tumours (0.48 +/- 0.12 and 0.43 +/- 0.08, respectively) was significantly lower than in the benign tumours (0.83 +/- 0.12 and 0.58 +/- 0.15, respectively). There were no significant differences in the utero-ovarian and intratumoural blood flow RIs when organic and functional tumours were compared. In all of the malignant tumours a colour signal was identified; it was centrally located in 90% of the cases. A colour signal was observed in 83% of the functional tumours, and in 52% of the organic benign tumours. In the benign tumours the colour signal was peripherally located in 98% of the cases. The variables for sensitivity, specificity, positive predictive value, negative predictive value and precision when organic tumours were diagnosed were 82.9, 65.5, 78.7, 71.4 and 76.1 for a sonographic index where the score was > or = 4; the values for the utero-ovarian RI were 48.4, 90.4, 88.4, 53.6 and 65.1 (cut-off 0.55 > or = RI > or = 0.90); the values for the intratumoural RI were 56.5, 68.6, 54.2, 70.6 and 63.8 (cut-off < or = 0.55). When malignancy was diagnosed these variables were 95.5, 82.2, 38.2, 99.4 and 83.6 for the sonographic index (score > or = 6); 90.9, 98.9, 90.0, 98.9 and 98.1 for the utero-ovarian RI (cut-off < or = 0.55) and 80.0, 66.7, 33.3, 94.1 and 69.0 for the intratumoural RI (cut-off < or = 0.50). The number of false positives diagnosed on the basis of the intratumoural RI decreased when cases that had been evaluated during the luteal phase were excluded from the study. Notwithstanding, the phase of the cycle does not seem to interfere with the diagnosis when utero-ovarian Doppler imaging is used., Conclusions: B-mode transvaginal ultrasonography and utero-ovarian Doppler velocimetry seem to complement each other to aid in differentiating between organic and functional ovarian tumours. Transvaginal ultrasonography and Doppler imaging provide good results for the diagnosis of malignancy, although the utero-ovarian RI reduces the number of false positives that occur with ultrasonography. Our group, therefore, believes that the combined use of these techniques is beneficial in the clinical d

Aim: To establish the proportion of patients with unstable angina in whom adequate anticoagulation is achieved using a standard regime of intravenous heparin., Method: A prospective series of 108 Emergency Department attendees over a six-month period with a clinical diagnosis of unstable angina for whom anticoagulation with heparin was prescribed were included in the study. The standard regime was a 5000 unit bolus followed by an intravenous infusion of 1000 units per hour (1200 units if the patient's weight was greater than 80 kg), with subsequent adjustments being made by reference to a nomogram. The activated partial thromboplastin time (APTT) was measured at six and 12 hours after treatment began. Two commonly used criteria for adequate heparinisation were compared: 1. APTT greater than 1.5 times control and 2. APTT in the range of 60-85 seconds., Results: There were valid data for 90 patients at six hours and 79 at 12 hours. Compared to the criterion for adequate anticoagulation of APTT greater than 1.5 times the control, 25% of patients were subtherapeutic at six hours and 12% at 12 hours. Compared to the criterion APTT greater than 60 seconds, 53% of patients were subtherapeutic at 6 hours and 47% at 12 hours. At 6 hours, 26% of patients were over-anticoagulated as defined as APTT greater than 85 seconds. This had reduced to 13% by 12 hours., Conclusions: In the context of recent research suggesting that an APTT of greater than 1.5 times the control is sufficient to reduce complications in unstable angina, our results demonstrate that a standard regime of heparinisation will achieve this goal in the majority of patients within 6 hours of starting heparin therapy. However, if an APTT of 60-85 seconds is the goal, this standard regime is inadequate.

Background: Acute pulmonary oedema (APO) is a frequent cause of respiratory failure and a common reason for presentation to emergency departments (EDs). To date, no paper has been published on the application of continuous positive airway pressure (CPAP) therapy for a large broad-based patient group., Aim: To report our experience with the use of CPAP in severe APO oedema, with particular reference to safety, intubation rates and impact on EDs' resources., Method: A retrospective chart review was undertaken of 75 patients with acute severe pulmonary oedema who were treated with adjuvant CPAP in an urban teaching hospital ED., Results: Three patients (4%) required subsequent endotracheal intubation and mechanical ventilation. The average duration of CPAP was 1.9 hours. Eighty nine per cent of patients experienced no adverse events while being treated with GPAP. Five patients failed to tolerate the tight fitting mask necessitating removal of CPAP, three patients experienced arrhythmias related to underlying cardiac disease and two patients experienced mild transient hypotension. Seventy one per cent of patients were discharged from the ED to general medical wards. The in-hospital mortality for patients treated with CPAP was 15%., Conclusion: This series has demonstrated that CPAP therapy delivered via a face mask for the treatment of acute severe APO is safe and effective when applied to a broad range of patients. We recommend the use of CPAP therapy for all suitable patients presenting in severe APO irrespective of age or underlying pulmonary disease.

Our aim was to study placental circulation during the first trimester of normal pregnancy. For this purpose, 108 single pregnancies from 4 to 15 gestational weeks were evaluated through conventional Doppler ultrasonography. The flow velocity waveforms from the retrochorionic arteries (spiral-radial arteries) and the umbilical artery were assessed using the peak systolic velocity, resistive index, and pulsatility index). Intervillous flow velocity waveform was evaluated from the maximum velocity. The earliest color signal from the retrochorionic circulation was registered at 4.5 weeks along with gestational sac visualization. The venous Doppler signal from the intervillous space and the Doppler signal from the umbilical artery were recorded with an embryo visible from the end of week 5 onward. The retrochorionic, intervillous, and umbilical peak systolic velocities increase, whereas the resistive and pulsatility indices decrease progressively during early pregnancy with a significant correlation with gestational age. Similarly, intervillous maximum velocity gradually increases throughout the first trimester of pregnancy. Despite some methodologic problems related to Doppler technology and the vessels studied color Doppler sonography appears to be an adequate tool to assess the physiologic changes in the placental circulation during early pregnancy.