bluuebunny/crossref_metadata_2025 huggingface dataset:
Optimizing the pre‐referral workup for gastroenterology and hepatology specialty care: consensus using the Delphi method
Authors: Ho Chanda K. , Boscardin Christy K. , Gleason Nathaniel , Collado Don , Terdiman Jonathan , Terrault Norah A. , Gonzales Ralph
Published: July 2015

Rationale, aims and objectives

Specialty care referrals have doubled in the last decade. Optimization of the pre‐referral workup by a primary care doctor can lead to a more efficient first specialty visit with the patient. Guidance regarding pre‐referral laboratory testing is a first step towards improving the specialty referral process. Our aim was to establish consensus regarding appropriate pre‐referral workup for common gastrointestinal and liver conditions.

Methods

The Delphi method was used to establish local consensus for recommending certain laboratory tests prior to specialty referral for 13 clinical conditions. Seven conditions from The University of Michigan outpatient referral guidelines were used as a baseline. An expert panel of three PCPs and nine gastroenterologists from three academic hospitals participated in three iterative rounds of electronic surveys. Each panellist ranked each test using a 5‐point Likert scale (strongly disagree to strongly agree). Local panellists could recommend additional tests for the initial diagnoses, and also recommended additional diagnoses needing guidelines: iron deficiency anaemia, abdominal pain, irritable bowel syndrome, fatty liver disease, liver mass and cirrhosis. Consensus was defined as ≥70% of experts scoring ≥4 (agree or strongly agree).

Results

Applying Delphi methodology to extrapolate externally developed referral guidelines for local implementation resulted in considerable modifications. For some conditions, many tests from the external group were eliminated by the local group (abdominal bloating; iron deficiency anaemia; irritable bowel syndrome). In contrast, for chronic diarrhoea, abnormal liver enzymes and viral hepatitis, all/most original tests were retained with additional tests added. For liver mass, fatty liver disease and cirrhosis, there was high concordance among the panel with few tests added or eliminated.

Conclusions

Consideration of externally developed referral guidelines using a consensus‐building process leads to significant local tailoring and adaption. Our next steps include implementation and dissemination of these guidelines and evaluating their impact on care efficiency in clinical practice.
Population genetics and evolution of the mangrove rivulus Kryptolebias marmoratus, the world's only self‐fertilizing hermaphroditic vertebrate
Authors: Avise J. C. , Tatarenkov A.
Published: July 2015

The mangrove rivulus, Kryptolebias marmoratus (Rivulidae, Cyprinodontiformes), is phylogenetically embedded within a large clade of oviparous (egg laying) and otherwise mostly gonochoristic (separate sex) killifish species in the circumtropical suborder Aplocheiloidei. It is unique in its reproductive mode: K. marmoratus is essentially the world's only vertebrate species known to engage routinely in self‐fertilization as part of a mixed‐mating strategy of selfing plus occasional outcrossing with gonochoristic males. This unique form of procreation has profound population‐genetic and evolutionary‐genetic consequences that are the subject of this review.
Generating precipitation ensembles for flood alert and risk management
Authors: Caseri A. , Javelle P. , Ramos M.H. , Leblois E.
Published: July 2015

Floods are major natural disasters that, in several occasions, can be responsible for life losses and severe economic damages. Flood forecasting and alert systems are needed to anticipate the arrival of these events and mitigate their impacts. They are particularly important for risk management and response in the nowcasting of flash floods. In this case, precipitation fields are crucial and is important to consider uncertainties coming from the observed precipitation fields used as input data to the system. One approach to take into account these uncertainties is to generate an ensemble of possible scenarios of observed precipitation. The aim of this study is to investigate the potential of a framework that applies a geostatistical conditional simulation method to generate an ensemble of precipitation fields that can be used as input to a distributed rainfall–runoff model to produce probabilistic flood alert maps. The Var region (southeastern France) and 17 events are used to validate the approach. Results show that the proposed method can be useful to generate realistic precipitation scenarios and, ultimately, to provide information on the probability of discharges exceeding critical flood thresholds. It can be a solution to combine information from radar fields and rain gauges to generate precipitation ensembles and quantify uncertainties in input data for hydrological modelling.
A real‐time pluvial flood forecasting system for Castries, St. Lucia
Authors: René J.‐R. , Djordjević S. , Butler D. , Mark O. , Henonin J. , Eisum N. , Madsen H.
Published: July 2015

In the last decade, real‐time flood forecasting has become a more feasible approach to reducing the impacts of flooding in urban areas. Two key tools in this context are high resolution hydrodynamic modelling in combination with accurate hydrological forcing. In some cases, when it is not possible to produce such accurate flood forecasts based on high resolution models and data, it may nevertheless be possible to use the resources currently available, accepting that there is a greater degree of uncertainty involved. This paper demonstrates the feasibility of a remotely controlled, real‐time, pluvial flood forecasting system for Castries, St. Lucia that utilises the limited data available locally. The results from the study suggest that although Global Forecast System (GFS) rainfall data may be considered coarse for urban applications, there is still a significant amount of skill and usability after it is postprocessed and used in combination with observed rainfall data. Evidence from the study also suggests that the use of images from different sources is invaluable for 2D overland model calibration and validation in urban areas. Conclusions from the study are potentially transferable to other sites in similar data‐scare and resource‐limited locations.
A large‐field polarisation‐resolved laser scanning microscope: applications to CARS imaging
Authors: DE VITO G. , CANTA A. , MARMIROLI P. , PIAZZA V.
Published: July 2015

Laser‐scanning imaging techniques are frequently used to probe the molecule spatial orientation in a sample of interest by exploiting selection rules depending on the polarisation of the excitation light. For the successful implementation of these techniques the precise control of the polarisation at the sample level is of fundamental importance. Polarisation distortions induced by the optical elements are often the main limitation factor for the maximum size of the field‐of‐view in polarisation‐resolved (PR) laser‐scanning microscopy, since for large scanning angles the polarisation distortions may mask the real sample structure. Here we shall demonstrate the implementation of large‐field‐of‐view PR microscopy and show PR CARS imaging of mouse spinal cord thanks to a careful design of the laser‐beam optical path. We shall show that this design leads to strongly suppressed distortions and quantify their effects on the final images. Although the focus of this work is on CARS imaging, we stress that the approaches described here can be successfully applied to a wide range of PR laser‐scanning techniques.
Genetic deletion of monoacylglycerol lipase leads to impaired cannabinoid receptor CB1R signaling and anxiety‐like behavior
Authors: Imperatore Roberta , Morello Giovanna , Luongo Livio , Taschler Ulrike , Romano Rosaria , De Gregorio Danilo , Belardo Carmela , Maione Sabatino , Di Marzo Vincenzo , Cristino Luigia
Published: July 2015

Endocannabinoids (eCB) are key regulators of excitatory/inhibitory neurotransmission at cannabinoid‐1‐receptor (CB1R)‐expressing axon terminals. The most abundant eCB in the brain, that is 2‐arachidonoylglycerol (2‐AG), is hydrolyzed by the enzyme monoacylglycerol lipase (MAGL), whose chronic inhibition in the brain was reported to cause CB1R desensitization. We employed the MAGL knock‐out mouse (MAGL−/−), a genetic model of congenital and sustained elevation of 2‐AG levels in the brain, to provide morphological and biochemical evidence for β‐arrestin2‐mediated CB1R desensitization in brain regions involved in the control of emotional states, that is, the prefrontal cortex (PFC), amygdala, hippocampus and cerebellar cortex. We found a widespread CB1R/β‐arrestin2 co‐expression in the mPFC, amygdala and hippocampus accompanied by impairment of extracellular signal‐regulated kinase signaling and elevation of vesicular glutamate transporter (VGluT1) at CB1R‐positive excitatory terminals in the mPFC, or vesicular GABA transporter (VGAT) at CB1R‐positive inhibitory terminals in the amygdala and hippocampus. The impairment of CB1R signaling in MAGL−/− mice was also accompanied by enhanced excitatory drive in the basolateral amygdala (BLA)‐mPFC circuit, with subsequent elevation of glutamate release to the mPFC and anxiety‐like and obsessive‐compulsive behaviors, as assessed by the light/dark box and marble burying tests, respectively. Collectively, these data provide evidence for a β‐arrestin2‐mediated desensitization of CB1R in MAGL−/− mice, with impact on the synaptic plasticity of brain circuits involved in emotional functions.

image

In this study, the authors provide evidence that congenitally enhanced endocannabinoid levels in the neuronal circuits underlying anxiety‐like behavioral states (mainly medial prefrontal cortex, amygdala and hippocampus) lead to CB1R desenistization and anxiety and depression. MAGL−/− mice, a model of congenital overactivity of the eCB system, exhibited a compensatory impairment of CB1R signaling in anxiety‐associated brain areas and a subsequent change in excitatory/inhibitory tone associated with altered score in the marble burying and light/dark box test, in concomitance with anxiety and depression behavior states. These findings may have potential relevance to the understanding of the neurochemical effects of chronic CB1R overstimulation in cannabis abusers.
Blue Revolution in a Commodity Frontier: Ecologies of Aquaculture and Agrarian Change in Laguna Lake,Philippines
Authors: Saguin Kristian
Published: July 2015

Aquaculture presents a radically different way of producing fish that aims to transcend the limitations of capture fisheries but that in turn creates new forms of agrarian and ecological transformations. Using the case of Laguna Lake, the paper probes how aquaculture production and corresponding agrarian transformations are inextricably tied to dynamics in capture fisheries in multiple ways. It emphasizes the fundamentally ecological nature of the relations between aquaculture and capture fisheries through a discussion of three interrelated features of agrarian change: commodity widening through the production of a commodity frontier, aquaculture producer strategies of working with materiality of biophysical nature, and the attendant consequences of these processes for agrarian configurations. By examining the appropriation of nature in commodity frontiers and situating relations between aquaculture and capture fisheries as historical‐geographical moments in commodity widening and deepening, the paper highlights the centrality of nature in agrarian change.
Quantitative comparison of topical aluminum salt solution efficacy for management of sweating: a randomized, controlled trial
Authors: Swary Jillian H , West Dennis P , Kakar Rohit , Ortiz Sara , Schaeffer Matthew R , Veledar Emir , Alam Murad
Published: July 2015

Background

There is a lack of studies objectively comparing the efficacy of topical antiperspirants in reducing sweat.

Objective

To objectively and quantitatively compare the efficacy of two aluminum salt solutions for the reduction of induced sweating.

Methods

A subject, rater, and statistician‐blinded, randomized, controlled trial. Nineteen subjects were exposed to a standardized heat challenge for 3 h. Topical agent A (20% aluminum chloride hexahydrate) was randomized to either axilla, and topical agent B (1% aluminum acetate) assigned to the contralateral side. A sauna suit induced sweating during three 30‐min heat intervals: (1) with no study agents (pre); (2) with both study agents, one on each side; and (3) after the agents were washed off (post). Sweat levels were measured by securing Whatman® filter paper to each axilla and measuring the paper weight after each heat interval. The difference in paper weight following each heat interval between Study Agent A and Study Agent B was measured by a gravimetric scale.

Results

Topical agent A had a significantly greater effect at reducing axillary sweating than B (P = 0.0002).

Conclusion

In a sweating simulation, 20% aluminum chloride hexahydrate quantitatively and objectively appeared to reduce sweat more effectively than 1% aluminum acetate.
Prognostic value of human epididymis protein 4 in endometrial cancer and its utility for surgical staging
Authors: Minar Lubos , Klabenesova Ivanka , Jandakova Eva , Zlamal Filip , Bienertova‐Vasku Julie
Published: July 2015

Aim

An optimal surgical staging in the group of patients with the high‐risk type of endometrial cancer is often limited by age and serious internal comorbidities. Therefore, in this study we focused on human epididymis protein 4 and its contribution to the preoperative differentiation of prognostically distinct groups of patients and to individualized surgical treatment as compared with cancer antigen (CA) 125 and imaging methods.

Material and Methods

The study included 115 patients with endometrioid adenocarcinoma diagnosed through endometrial biopsy. Before the final operation, blood sampling was performed for the determination of human epididymis protein 4 (HE4) and CA125 levels. Serum levels of both biomarkers were analyzed in relation to individual prognostic factors (stage of disease, depth of myometrial invasion, tumor grade, risk type of disease).

Results

In the case of HE4, we demonstrated a statistically significant difference (P < 0.001) between patients with low and high risk of the disease. In our model, achieving the maximum sum of sensitivity and specificity, HE4 shows a sensitivity of 72.4% and a specificity of 75.4% for the cut‐off 76.5 pmol/L and is a better predictor in distinguishing the high‐risk patients than CA125 (area under the curve 0.77 for HE vs 0.71 for CA125).

Conclusion

HE4 is a marker that could complement the findings of imaging techniques and that may be useful in decision‐making on how to individualize surgical staging. The possibility of its introduction as an independent marker in routine practice remains, at the moment however, limited. The optimal cut‐off for HE4 has not been established yet and further studies are needed.
Efficacy of multiple microwave endometrial ablation technique for menorrhagia resulting from adenomyosis
Authors: Nakamura Kohei , Nakayama Kentaro , Ishikawa Masako , Katagiri Hiroshi , Katagiri Atsuko , Ishibashi Tomoka , Sato Emi , Asakawa Yasuyuki , Kyo Satoru
Published: July 2015

Aim

Conventional microwave endometrial ablation (MEA) can be insufficient to control menorrhagia resulting from adenomyosis. We compared the standard single ablation technique with multiple MEA – repeating ablation three times in the same region – in patients with adenomyosis and menorrhagia.

Material and Methods

We performed single MEA in 18 patients and multiple MEA in seven patients between 2007 and 2013. We compared the efficacy of single and multiple MEA using a visual analog scale (VAS) for menorrhagia, dysmenorrhea, and patient satisfaction. We also evaluated the incidence of menorrhagia recurrence, amenorrhea, and procedure complications in relation to patients' clinical factors.

Results

VAS scores for improved menorrhagia and patient satisfaction were significantly higher in the multiple MEA group than in the single MEA group; however, the operative time was longer in the multiple‐treatment group. There were no statistical differences between groups in hemoglobin levels, VAS improvement for dysmenorrhea, menorrhagia recurrence, frequency of complications, or amenorrhea rate.

Conclusion

Multiple MEA successfully controls menorrhagia from adenomyosis and achieves a higher satisfaction rate than single MEA.
Continuous and long‐term treatment is more important than dosage for the protective effect of thiazide use on bone metabolism and fracture risk
Authors: Kruse C. , Eiken P. , Vestergaard P.
Published: July 2015

Background

Data from observational studies have suggested that thiazide diuretics protect against fractures. Few studies have investigated time frames from initiation of treatment to fracture occurrence.

Objective

To evaluate the time to spinal, hip, femur, wrist and upper extremity fracture occurrence before and after thiazide exposure.

Methods

A matched retrospective cohort study of patient information from national Danish patient databases was conducted. Patients with reimbursed prescriptions for noncompounded thiazide diuretics with potassium supplementation (Anatomical Therapeutic Chemical classification system code C03AB) between 1996 and 2011 were matched with nonexposed control subjects by date of birth and gender. Weekly odds ratios (ORs) of fracture occurrence and total incidence rates (IRs) and incidence rate ratios (IRRs) of fracture risk were calculated for the periods before treatment initiation, weeks 1–42 and weeks 43–780.

Results

A total of 1 602 141 ‘thiazide exposure periods’ (46 8271 individuals) and 1 530 233 ‘nonexposure periods’ (655 399 individuals) were included in the analysis. Thiazide use was associated with factors of increased de novo fracture risk. Weekly adjusted fracture risk between exposure and nonexposure was increased prior to commencing thiazide therapy, further increasing from weeks 1–42 weeks and then decreasing gradually from weeks 43–780. There was a decreasing trend in total age‐adjusted risk during these periods: IRR [95% confidence interval 1.44 [1.42; 1.47], 1.27 [1.24; 1.29] and 1.14 [1.11; 1.18], respectively. Prescription patterns showed several treatment breaks amongst thiazide users.

Conclusions

It appears that thiazides reduce the background risk of fracture that is increased prior to commencing therapy. Long duration and continuity of thiazide exposure seems to be important to obtain this protective effect on fracture risk, but we have found in this study that this approach is not always used in clinical practice.
A comparison of postrelease survival parameters between single and mass stranded delphinids from Cape Cod, Massachusetts, U.S.A.
Authors: Sharp Sarah M. , Harry Charles T. , Hoppe Jane M. , Moore Kathleen M. , Niemeyer Misty E. , Robinson Ian , Rose Kathryn S. , Sharp W. Brian , Landry Scott , Richardson Jessica , Moore Michael J.
Published: July 2015

The viability of healthy single stranded dolphins as immediate release candidates has received little attention. Responders have been reluctant to release lone delphinids due to their social needs, even when they pass the same health evaluations as mass stranded animals. This study tracked postrelease success of 34 relocated and released satellite tagged delphinids from single and mass strandings. Three postrelease survival parameters (transmission duration, swim speed, and daily distance) were examined to evaluate whether they differed among single stranded/single released (SS/SR), mass stranded/single released (MS/SR), or mass stranded/mass released (MS/MR) dolphin groups. Comparisons were also made between healthy and borderline release candidates. Satellite tags transmitted for a mean of 21.2 d (SD = 19.2, range = 1–79), daily distance traveled was 42.0 km/d (11.25, 20.96–70.72), and swim speed was 4.3 km/h (1.1, 2.15–8.54). Postrelease parameters did not differ between health status groups, however, SS/SR dolphins transmitted for a shorter mean duration than MS/MR and MS/SR groups. Postrelease vessel‐based surveys confirmed conspecific group location for two healthy, MS/SR dolphins. Overall, these results support the potential to release healthy stranded single delphinids; however, further refinement of health assessment protocols for these challenging cases is needed.
The use of colonic and anorectal high‐resolution manometry and its place in clinical work and in research
Authors: Dinning P. G. , Carrington E. V. , Scott S. M.
Published: July 2015

Background

In the esophagus, high‐resolution manometry (HRM) has become a standard diagnostic tool in the investigation of suspected motility disorders. However, at the opposite end of the digestive tract (i.e., the colon and anorectum), the use of HRM still remains in its infancy, with relatively few published studies in the scientific literature. Further, the clinical utility of those studies that have been performed is largely undetermined.

Purpose

This review assesses all of the HRM studies published to date from both the colon and anorectum, explores the catheter types used, and attempts to determine the worth of HRM over traditional ‘low‐resolution’ recordings from the same regions. Ultimately, this review addresses whether HRM currently provides information that will benefit patient diagnosis and treatment.
An overview of the efficacy of resveratrol in the management of ischemic heart disease
Authors: Raj Pema , Zieroth Shelley , Netticadan Thomas
Published: July 2015

Ischemic heart disease is a leading cause of cardiac dysfunction and subsequent morbidity and mortality around the world. New therapies are required to complement or enhance the existing treatment regimen for the management of ischemic heart disease–related clinical complications. In this regard, compounds derived from natural sources have recently gained attention for their cardioprotective properties. In particular, the potential of food‐derived compounds that exhibit medicinal properties (termed nutraceuticals) appears promising, an example being the plant polyphenol resveratrol. In the past two decades, many preclinical and a few pilot clinical studies have shown that resveratrol is beneficial in protecting against cardiovascular disease. In this short review, we will discuss current evidence on the efficacy of resveratrol in preventing or reversing deleterious effects on the heart in the setting of ischemic heart disease.
Effects of peripheral blood mononuclear cells on Haemonchus contortus larval motility in vitro
Authors: Holt R. M. , Shepherd E. A. , Ammer A. G. , Bowdridge S. A.
Published: July 2015

The objective of this experiment was to determine effects of peripheral blood mononuclear cells (PBMC), derived from parasite‐resistant St. Croix (STC) and parasite‐susceptible Suffolk (SF) sheep, on motility of Haemonchus contortus L3 stage larvae in vitro. Peripheral blood mononuclear cells were collected from 10 lambs of each breed, 5 naïve and 5 which had received a priming infection with H. contortus. Larval motility was quantified using a MIF Nikon Sweptfield microscope and NIS Elements AR software, and measurements included path length (PL) (μm), velocity (VEL) (μm/s) and acceleration (ACC) (μm/s2). After 18 h of incubation, PL and VEL were greatest in larvae cultured with SF‐derived PMBC and were significantly different from all other groups (P < 0·01). No difference was observed in PL or VEL between larvae exposed to naïve or primed STC‐derived PBMC and primed‐SF PBMC. Differences in ACC were detected between larvae cultured with primed STC‐derived PBMC (10·91 μm/s2) and naïve SF‐derived PBMC (45·7 μm/s2) (P = 0·035). These data indicate an innate ability of STC‐derived PBMC to severely inhibit larval motility.
Augmenting the Deliberative Method for Ranking Risks
Authors: Susel Irving , Lasley Trace , Montezemolo Mark , Piper Joel
Published: July 2015

The Department of Homeland Security (DHS) characterized and prioritized the physical cross‐border threats and hazards to the nation stemming from terrorism, market‐driven illicit flows of people and goods (illegal immigration, narcotics, funds, counterfeits, and weaponry), and other nonmarket concerns (movement of diseases, pests, and invasive species). These threats and hazards pose a wide diversity of consequences with very different combinations of magnitudes and likelihoods, making it very challenging to prioritize them. This article presents the approach that was used at DHS to arrive at a consensus regarding the threats and hazards that stand out from the rest based on the overall risk they pose. Due to time constraints for the decision analysis, it was not feasible to apply multiattribute methodologies like multiattribute utility theory or the analytic hierarchy process. Using a holistic approach was considered, such as the deliberative method for ranking risks first published in this journal. However, an ordinal ranking alone does not indicate relative or absolute magnitude differences among the risks. Therefore, the use of the deliberative method for ranking risks is not sufficient for deciding whether there is a material difference between the top‐ranked and bottom‐ranked risks, let alone deciding what the stand‐out risks are. To address this limitation of ordinal rankings, the deliberative method for ranking risks was augmented by adding an additional step to transform the ordinal ranking into a ratio scale ranking. This additional step enabled the selection of stand‐out risks to help prioritize further analysis.
Practical Case Application for Stress Model Validation and Enhancement by Means of Metrological Tools
Authors: Gutiérrez R. , Ramírez M. , Olmeda E. , Díaz V.
Published: July 2015

Models consider ideal and simplified situations that will never be met in the real case. The process of comparing model predictions and experimental observation is in the basis of scientific research. This comparison is however complicated because of the uncertainties of the model input data and the difficulty to control the accuracy of the tests and to obtain a significant statistical sampling. Moreover, there isn't yet a consensus on a validation parameter. This paper presents a three‐step validation procedure that allows quantifying the application limits of a two‐dimensional stress model in a three‐dimensional situation. A global uncertainty model is calculated comprising the uncertainty of the model and also the uncertainty coming from the experimental results. The EN number, a statistical magnitude for interlaboratory comparisons, is used to analyse the compatibility between the experimental and theoretical results. Finally, a bootstrapping method is proposed to calculate the coverage interval of the sampling and determine if new experiments should be carried out. Numerical results of this new validation procedure are provided for the case under study. It is also demonstrated that the computed uncertainty budget can be a useful tool to enhance the two‐dimensional model by enlarging its uncertainty limits.
The Second Zambian National Tuberculosis Drug Resistance survey – a comparison of conventional and molecular methods
Authors: Kapata Nathan , Mbulo Grace , Cobelens Frank , de Haas Petra , Schaap Ab , Mwamba Pike , Mwanza Winnie , Muvwimi Mweemba , Muyoyeta Monde , Moyo Maureen , Mulenga Lutinala , Grobusch Martin P. , Godfrey‐Faussett Peter , Ayles Helen
Published: July 2015

Objective

The prevalence of MDR‐TB in Zambia was estimated to be 1.8% in 2001. A second drug resistance survey was conducted in 2008 to determine trends; the use of the Genotype MTBDRplus assay was applied to compare results to the gold standard.

Method

A two‐stage cluster sampling, with health facilities as primary sampling units. Processed sputum specimens were inoculated on solid media for culture; heat‐inactivated bacterial suspensions from sputum samples were tested on a commercial line probe assay for the identification of rifampicin and isoniazid resistance.

Results

A total of 917 patients with TB were enrolled and 883 (96.3%) analysed. A total of 574 (65%) had LJ results and 824 (93.3%) had results from MTBDRplus assay. The median age was 32, and 63.3% were males. MDR‐TB according to LJ‐based DST was 1.1% (CI 0.1–2.4) whereas according to MDTBDRplus assay was 1.6% (CI 0.6–2.6). Isoniazid monoresistance in new cases was 2.4% (CI 0.613–4.26) based on LJ results and 5.0% (CI 3.2–6.7) based on the MTBDRplus; in retreatment cases, it was 4.4% (CI 0.3–8.6) and 2.40% (CI <0.1–5.1) on LJ and MTBDRplus, respectively. Rifampicin monoresistance in new cases was 0.1% (CI <0.1–0.4) based on LJ and 0.6% (CI 0.01–1.1) based on the MTBDRplus; in retreatment cases, it was 0% (CI 0–3.8) and 1.8% (CI <0.1–4.0) on LJ and MTBDRplus, respectively. There were no XDR‐TB cases found and no association between MDR‐TB and HIV.

Conclusion

There was no increase in MDR‐TB prevalence in Zambia from 2001 to 2008; results from the two methods were similar. Molecular methods were quicker and simpler to use.
Alginate Microspheroid Encapsulation and Delivery of MG-63 Cells Into Polycaprolactone Scaffolds: A New Biofabrication Approach for Tissue Engineering Constructs
Authors: Narayanan Lokesh K. , Kumar Arun , Tan Zhuo (George) , Bernacki Susan , Starly Binil , Shirwaiker Rohan A.
Published: July 2015

Scaffolds play an important role in tissue engineering by providing structural framework and a surface for cells to attach, proliferate, and secrete extracellular matrix (ECM). In order to enable efficient tissue formation, delivering sufficient cells into the scaffold three-dimensional (3D) matrix using traditional static and dynamic seeding methods continues to be a critical challenge. In this study, we investigate a new cell delivery approach utilizing deposition of hydrogel-cell encapsulated microspheroids into polycaprolactone (PCL) scaffolds to improve the seeding efficiency. Three-dimensional-bioplotted PCL constructs (0 deg/90 deg lay down, 284 ± 6 μm strand width, and 555 ± 8 μm strand separation) inoculated with MG-63 model bone cells encapsulated within electrostatically generated calcium-alginate microspheroids (Ø 405 ± 13 μm) were evaluated over seven days in static culture. The microspheroids were observed to be uniformly distributed throughout the PCL scaffold cross section. Encapsulated cells remained viable within the constructs over the test interval with the highest proliferation noted at day 4. This study demonstrates the feasibility of the new approach and highlights the role and critical challenges to be addressed to successfully utilize 3D-bioprinting for microencapsulated cell delivery.
Novel techniques and devices for in-situ film coatings of long, small diameter tubes or elliptical and other surface contours
Authors: Hershcovitch Ady , Blaskiewicz Michael , Brennan Joesph Michael , Fischer Wolfram , Liaw Chong-Jer , Meng Wuzhang , Todd Robert , Custer Art , Dingus Aaron , Erickson Mark , Jamshidi Nader , Poole Henry Joe
Published: July 2015

Devices and techniques that can, via physical vapor deposition, coat various surface contours or very long small aperture pipes, are described. Recently, a magnetron mole was developed in order to in-situ coat accelerator tube sections of the Brookhaven National Lab relativistic heavy ion collider that have 7.1 cm diameter with access points that are 500 m apart, for copper coat the accelerator vacuum tube in order to alleviate the problems of unacceptable ohmic heating and of electron clouds. A magnetron with a 50 cm long cathode was designed fabricated and successfully operated to copper coat a whole assembly containing a full-size, stainless steel, cold bore, of the accelerator magnet tubing connected to two types bellows, to which two additional pipes made of accelerator tubing were connected. The magnetron is mounted on a carriage with spring loaded wheels that successfully crossed bellows and adjusted for variations in vacuum tube diameter, while keeping the magnetron centered. Electrical power and cooling water were fed through a cable bundle. The umbilical cabling system, which is enclosed in a flexible braided metal sleeve, is driven by a motorized spool. To increase cathode lifetime, movable magnet package was developed, and thickest possible cathode was made, with a rather challenging target to substrate distance of less than 1.5 cm. Optimized process to ensure excellent adhesion was developed. Coating thickness of 10 μm Cu passed all industrial tests and even exceeded maximum capability of a 12 kg pull test fixture. Room temperature radio frequency (RF) resistivity measurement indicated that 10 μm Cu coated stainless steel accelerator tube has conductivity close to copper tubing. Work is in progress to repeat the RF resistivity measurement at cryogenic temperatures. Over 20 years ago, a device using multiaxis robotic manipulators controlling separate robotic assemblies resulted in nine-axes of motion combined with conformal shape of the cathodes that can adapt to various curved surface contours was developed and successfully used for depositing optical coating on aircraft canopies. The techniques can be utilized for in situ coating of elliptical and other surface contour RF cavities and long beam pipes with thick superconducting films. Plans are to incorporate ion assisted deposition in those techniques for attaining dense, adherent and defect free coatings.
Focused-ion beam patterning of organolead trihalide perovskite for subwavelength grating nanophotonic applications
Authors: Alias Mohd Sharizal , Dursun Ibrahim , Shi Dong , Saidaminov Makhsud Ismatboevich , Diallo Elhadj Marwane , Priante Davide , Ng Tien Khee , Bakr Osman Mohammed , Ooi Boon Siew
Published: July 2015

The coherent amplified spontaneous emission and high photoluminescence quantum efficiency of organolead trihalide perovskite have led to research interest in this material for use in photonic devices. In this paper, the authors present a focused-ion beam patterning strategy for methylammonium lead tribromide (MAPbBr3) perovskite crystal for subwavelength grating nanophotonic applications. The essential parameters for milling, such as the number of scan passes, dwell time, ion dose, ion current, ion incident angle, and gas-assisted etching, were experimentally evaluated to determine the sputtering yield of the perovskite. Based on our patterning conditions, the authors observed that the sputtering yield ranged from 0.0302 to 0.0719 μm3/pC for the MAPbBr3 perovskite crystal. Using XeF2 for the focused-ion beam gas-assisted etching, the authors determined that the etching rate was reduced to between 0.40 and 0.97, depending on the ion dose, compared with milling with ions only. Using the optimized patterning parameters, the authors patterned binary and circular subwavelength grating reflectors on the MAPbBr3 perovskite crystal using the focused-ion beam technique. Based on the computed grating structure with around 97% reflectivity, all of the grating dimensions (period, duty cycle, and grating thickness) were patterned with nanoscale precision (>±3 nm), high contrast, and excellent uniformity. Our results provide a platform for utilizing the focused-ion beam technique for fast prototyping of photonic nanostructures or nanodevices on organolead trihalide perovskite.
Tiny built-in lasers light up living cells
Authors: Cho Adrian
Published: July 2015

Technique could revolutionize tracking of individual cells.
Two leaps forward for robot locomotion
Authors: Vella Dominic
Published: July 2015

Biomimetic robots can jump on land and on water

[Also see Report by

Koh
et al.

]
A p53-regulated immune checkpoint relevant to cancer
Authors: Zitvogel Laurence , Kroemer Guido
Published: July 2015

The tumor suppressor p53 may influence the ability of cancer cells to escape immune detection

[Also see Research Article by

Yoon
et al.

]
Advances in the Diagnosis of Human Schistosomiasis
Authors: Weerakoon Kosala G. A. D. , Gobert Geoffrey N. , Cai Pengfei , McManus Donald P.
Published: July 2015

Schistosomiasis is a major neglected tropical disease that afflicts more than 240 million people, including many children and young adults, in the tropics and subtropics. The disease is characterized by chronic infections with significant residual morbidity and is of considerable public health importance, with substantial socioeconomic impacts on impoverished communities. Morbidity reduction and eventual elimination through integrated intervention measures are the focuses of current schistosomiasis control programs. Precise diagnosis of schistosome infections, in both mammalian and snail intermediate hosts, will play a pivotal role in achieving these goals. Nevertheless, despite extensive efforts over several decades, the search for sensitive and specific diagnostics for schistosomiasis is ongoing. Here we review the area, paying attention to earlier approaches but emphasizing recent developments in the search for new diagnostics for schistosomiasis with practical applications in the research laboratory, the clinic, and the field. Careful and rigorous validation of these assays and their cost-effectiveness will be needed, however, prior to their adoption in support of policy decisions for national public health programs aimed at the control and elimination of schistosomiasis.
Adenovirus E4-ORF3 Targets PIAS3 and Together with E1B-55K Remodels SUMO Interactions in the Nucleus and at Virus Genome Replication Domains
Authors: Higginbotham Jennifer M. , O'Shea Clodagh C.
Published: July 2015

Adenovirus E4-ORF3 and E1B-55K converge in subverting critical overlapping cellular pathways to facilitate virus replication. Here, we show that E1B-55K and E4-ORF3 induce sumoylation and the assembly of SUMO2/3 viral genome replication domains. Using a conjugation-deficient SUMO2 construct, we demonstrate that SUMO2/3 is recruited to E2A viral genome replication domains through noncovalent interactions. E1B-55K and E4-ORF3 have critical functions in inactivating MRN and ATM to facilitate viral genome replication. We show that ATM kinase inhibitors rescue ΔE1B-55K/ΔE4-ORF3 viral genome replication and that the assembly of E2A domains recruits SUMO2/3 independently of E1B-55K and E4-ORF3. However, the morphology and organization of SUMO2/3-associated E2A domains is strikingly different from that in wild-type Ad5-infected cells. These data reveal that E1B-55K and E4-ORF3 specify the nuclear compartmentalization and structure of SUMO2/3-associated E2A domains, which could have important functions in viral replication. We show that E4-ORF3 specifically targets and sequesters the cellular E3 SUMO ligase PIAS3 but not PIAS1, PIAS2, or PIAS4. The assembly of E4-ORF3 into a multivalent nuclear matrix is required to target PIAS3. In contrast to MRN, PIAS3 is targeted by E4-ORF3 proteins from disparate adenovirus subgroups. Our studies reveal that PIAS3 is a novel and evolutionarily conserved target of E4-ORF3 in human adenovirus infections. Furthermore, we reveal that viral proteins not only disrupt but also usurp SUMO2/3 to transform the nucleus and assemble novel genomic domains that could facilitate pathological viral replication.

IMPORTANCE
SUMO is a key posttranslational modification that modulates the function, localization, and assembly of protein complexes. In the ever-escalating host-pathogen arms race, viruses have evolved strategies to subvert sumoylation. Adenovirus is a small DNA tumor virus that is a global human pathogen and key biomedical agent in basic research and therapy. We show that adenovirus infection induces global changes in SUMO localization and conjugation. Using virus and SUMO mutants, we demonstrate that E1B-55K and E4-ORF3 disrupt and usurp SUMO2/3 interactions to transform the nucleus and assemble highly structured and compartmentalized viral genome domains. We reveal that the cellular E3 SUMO ligase PIAS3 is a novel and conserved target of E4-ORF3 proteins from disparate adenovirus subgroups. The induction of sumoylation and SUMO2/3 viral replication domains by early viral proteins could play an important role in determining the outcome of viral infection.
Vaccinia Virus B1 Kinase Is Required for Postreplicative Stages of the Viral Life Cycle in a BAF-Independent Manner in U2OS Cells
Authors: Jamin Augusta , Ibrahim Nouhou , Wicklund April , Weskamp Kaitlin , Wiebe Matthew S.
Published: July 2015

The vaccinia virus B1R gene encodes a highly conserved protein kinase that is essential for the poxviral life cycle. As demonstrated in many cell types, B1 plays a critical role during viral DNA replication when it inactivates the cellular host defense effector barrier to autointegration factor (BAF or BANF1). To better understand the role of B1 during infection, we have characterized the growth of a B1-deficient temperature-sensitive mutant virus (Cts2 virus) in U2OS osteosarcoma cells. In contrast to all other cell lines tested to date, we found that in U2OS cells, Cts2 viral DNA replication is unimpaired at the nonpermissive temperature. However, the Cts2 viral yield in these cells was reduced more than 10-fold, thus indicating that B1 is required at another stage of the vaccinia virus life cycle. Our results further suggest that the host defense function of endogenous BAF may be absent in U2OS cells but can be recovered through either overexpression of BAF or fusion of U2OS cells with mouse cells in which the antiviral function of BAF is active. Interestingly, examination of late viral proteins during Cts2 virus infection demonstrated that B1 is required for optimal processing of the L4 protein. Finally, execution point analyses as well as electron microscopy studies uncovered a role for B1 during maturation of poxviral virions. Overall, this work demonstrates that U2OS cells are a novel model system for studying the cell type-specific regulation of BAF and reveals a role for B1 beyond DNA replication during the late stages of the viral life cycle.

IMPORTANCE
The most well characterized role for the vaccinia virus B1 kinase is to facilitate viral DNA replication by phosphorylating and inactivating BAF, a cellular host defense responsive to foreign DNA. Additional roles for B1 later in the viral life cycle have been postulated for decades but are difficult to examine directly due to the importance of B1 during DNA replication. Here, we demonstrate that in U2OS cells, a B1 mutant virus escapes the block in DNA replication observed in other cell types and, instead, this mutant virus exhibits impaired late protein accumulation and incomplete maturation of new virions. These data provide the clearest evidence to date that B1 is needed for multiple critical junctures in the poxviral life cycle in a manner that is both dependent on and independent of BAF.
RIG-I Mediates an Antiviral Response to Crimean-Congo Hemorrhagic Fever Virus
Authors: Spengler Jessica R. , Patel Jenish R. , Chakrabarti Ayan K. , Zivcec Marko , García-Sastre Adolfo , Spiropoulou Christina F. , Bergeron Éric
Published: July 2015

In the cytoplasm, the retinoic acid-inducible gene I (RIG-I) senses the RNA genomes of several RNA viruses. RIG-I binds to viral RNA, eliciting an antiviral response via the cellular adaptor MAVS. Crimean-Congo hemorrhagic fever virus (CCHFV), a negative-sense RNA virus with a 5′-monophosphorylated genome, is a highly pathogenic zoonotic agent with significant public health implications. We found that, during CCHFV infection, RIG-I mediated a type I interferon (IFN) response via MAVS. Interfering with RIG-I signaling reduced IFN production and IFN-stimulated gene expression and increased viral replication. Immunostimulatory RNA was isolated from CCHFV-infected cells and from virion preparations, and RIG-I coimmunoprecipitation of infected cell lysates isolated immunostimulatory CCHFV RNA. This report serves as the first description of a pattern recognition receptor for CCHFV and highlights a critical signaling pathway in the antiviral response to CCHFV.

IMPORTANCE
CCHFV is a tick-borne virus with a significant public health impact. In order for cells to respond to virus infection, they must recognize the virus as foreign and initiate antiviral signaling. To date, the receptors involved in immune recognition of CCHFV are not known. Here, we investigate and identify RIG-I as a receptor involved in initiating an antiviral response to CCHFV. This receptor initially was not expected to play a role in CCHFV recognition because of characteristics of the viral genome. These findings are important in understanding the antiviral response to CCHFV and support continued investigation into the spectrum of potential viruses recognized by RIG-I.
Fitness-Balanced Escape Determines Resolution of Dynamic Founder Virus Escape Processes in HIV-1 Infection
Authors: Sunshine Justine E. , Larsen Brendan B. , Maust Brandon , Casey Ellie , Deng Wenje , Chen Lennie , Westfall Dylan H. , Kim Moon , Zhao Hong , Ghorai Suvankar , Lanxon-Cookson Erinn , Rolland Morgane , Collier Ann C. , Maenza Janine , Mullins James I. , Frahm Nicole
Published: July 2015

To understand the interplay between host cytotoxic T-lymphocyte (CTL) responses and the mechanisms by which HIV-1 evades them, we studied viral evolutionary patterns associated with host CTL responses in six linked transmission pairs. HIV-1 sequences corresponding to full-length p17 and p24
gag
were generated by 454 pyrosequencing for all pairs near the time of transmission, and seroconverting partners were followed for a median of 847 days postinfection. T-cell responses were screened by gamma interferon/interleukin-2 (IFN-γ/IL-2) FluoroSpot using autologous peptide sets reflecting any Gag variant present in at least 5% of sequence reads in the individual's viral population. While we found little evidence for the occurrence of CTL reversions, CTL escape processes were found to be highly dynamic, with multiple epitope variants emerging simultaneously. We found a correlation between epitope entropy and the number of epitope variants per response (
r
= 0.43;
P
= 0.05). In cases in which multiple escape mutations developed within a targeted epitope, a variant with no fitness cost became fixed in the viral population. When multiple mutations within an epitope achieved fitness-balanced escape, these escape mutants were each maintained in the viral population. Additional mutations found to confer escape but undetected in viral populations incurred high fitness costs, suggesting that functional constraints limit the available sites tolerable to escape mutations. These results further our understanding of the impact of CTL escape and reversion from the founder virus in HIV infection and contribute to the identification of immunogenic Gag regions most vulnerable to a targeted T-cell attack.

IMPORTANCE
Rapid diversification of the viral population is a hallmark of HIV-1 infection, and understanding the selective forces driving the emergence of viral variants can provide critical insight into the interplay between host immune responses and viral evolution. We used deep sequencing to comprehensively follow viral evolution over time in six linked HIV transmission pairs. We then mapped T-cell responses to explore if mutations arose due to adaption to the host and found that escape processes were often highly dynamic, with multiple mutations arising within targeted epitopes. When we explored the impact of these mutations on replicative capacity, we found that dynamic escape processes only resolve with the selection of mutations that conferred escape with no fitness cost to the virus. These results provide further understanding of the complicated viral-host interactions that occur during early HIV-1 infection and may help inform the design of future vaccine immunogens.
Cross section for (nearly) everyone
Authors: Cavinato Michela , Giliberti Marco , Perotti Luca
Published: July 2015

Both total and differential cross sections for the reflection of light beams off finite solid surfaces obtained by the rotation of a generic derivable convex function are presented. The calculations are developed using elementary notions of mathematics and are therefore suitable for introductory undergraduate university physics courses. Three particular cases are discussed as examples of the general procedure and of the possible considerations about cross sections in general and the physical properties of specific systems.