Egyptian Canopic Jars at the Crossroad of Medicine and Archaeology: Overview of 100 Years of Research and Future Scientific Expectations.
Ancient Egyptian human remains have been of interest in the fields of both medical and Egyptological research for decades. However, canopic jar holders for internal organs (liver, lungs, stomach, intestines) of Egyptian mummies appear to be but a very occasional source of data for such investigations. The few medical approaches focusing on the content of these jars are summarized and listed according to pathogens and diseases to give a structured overview of this field of study. An extensive search of the literature has been conducted from different bibliographic databases with a total of n = 26 studies found. The majority of diseases found consisted of infectious diseases and internal medicine conditions such as schistosomiasis or emphysema. These are just 2 examples of many that, instead of primarily affecting bone, muscle or skin, specifically target internal organs. Hence, a better understanding of the evolution of diseases that still affect mankind could be gained. In conclusion, this reassessment shows that canopic jars represent a highly underestimated source for histological, radiological and ancient DNA examination of Ancient Egyptian remains and should, thus, be more and more brought back into the focus of retrospective medical research.


The utilization of standardized order sets using AASLD guidelines for patients with suspected cirrhosis and acute gastrointestinal bleeding.
To improve the adherence to AASLD (American Association for the Study of Liver Diseases) guidelines for variceal bleeding, we developed and implemented standardized order sets for gastrointestinal bleeding in our hospital on October 1, 2009. We performed medical record reviews of hospitalized patients with gastrointestinal bleeding with suspected cirrhosis from October 2009 to October 2010 to determine the use of octreotide, prophylactic antibiotics, and endoscopy. We reviewed 300 Medical records and identified 26 patients with suspected cirrhosis and gastrointestinal bleeding who had adequate information to determine whether or not the order set was used. Antibiotic was used in 76% of patients, octreotide was used in 76% of patients, and upper endoscopy was completed in 94% of patients within 24 hours. The use of antibiotics was higher than that used in historical controls in our hospital. Implementation of standardized order sets appears to have improved adherence to standard recommendations. However, larger studies with longer follow-ups are needed to evaluate this effect on clinical outcomes and cost of care.


Compliance to Prenatal Iron and Folic Acid Supplement Use in Relation to Low Birth Weight in Lilongwe, Malawi.
Prenatal iron and folic acid (IFA) supplements are offered free to all pregnant women in Malawi to reduce maternal anemia and improve birth outcomes. We investigated the association between self-reported compliance to IFA intake and risk of low birth weight (LBW). Pregnant women who attended Bwaila Maternity Wing of Lilongwe District Hospital for delivery were recruited (n = 220). We used a questionnaire to collect self-reported information on IFA use and maternal sociodemographic data. Before delivery, blood samples for maternal hemoglobin (Hb) and folate status, and upon delivery, birth weight, and other newborn anthropometrics were measured. We used multivariable logistic regression to determine risk of LBW by prenatal IFA intake. The self-reported number of IFA pills taken during pregnancy was positively associated with Hb, but not serum and RBC folate concentration: <45, 45⁻89 and ≥90 pills taken corresponded with mean (SD) Hb 10.7 (1.6), 11.3 (1.8), and 11.7 (1.6) g/dL, respectively (p = 0.006). The prevalence of LBW was 20.1%, 13.5% and 5.6% for those who reported taking IFA pills <45, 45⁻89, and ≥90 pills, respectively (p = 0.027). Taking >60 IFA pills reduced risk of LBW delivery (OR (95% CI) = 0.15 (0.03⁻0.70), p = 0.033) than taking ≤30 pills. Self-reported compliance to IFA use is valid for assessing prenatal supplement program in Malawi, especially Hb status, and can reduce the rate of LBW.


"Mind the trap": mindfulness practice reduces cognitive rigidity.
Two experiments examined the relation between mindfulness practice and cognitive rigidity by using a variation of the Einstellung water jar task. Participants were required to use three hypothetical jars to obtain a specific amount of water. Initial problems were solvable by the same complex formula, but in later problems ("critical" or "trap" problems) solving was possible by an additional much simpler formula. A rigidity score was compiled through perseverance of the complex formula. In Experiment 1, experienced mindfulness meditators received significantly lower rigidity scores than non-meditators who had registered for their first meditation retreat. Similar results were obtained in randomized controlled Experiment 2 comparing non-meditators who underwent an eight meeting mindfulness program with a waiting list group. The authors conclude that mindfulness meditation reduces cognitive rigidity via the tendency to be "blinded" by experience. Results are discussed in light of the benefits of mindfulness practice regarding a reduced tendency to overlook novel and adaptive ways of responding due to past experience, both in and out of the clinical setting.


Evidence-based value of prophylactic drainage in gastrointestinal surgery: a systematic review and meta-analyses.
To determine the evidence-based value of prophylactic drainage in gastrointestinal (GI) surgery. An electronic search of the Medline database from 1966 to 2004 was performed to identify articles comparing prophylactic drainage with no drainage in GI surgery. The studies were reviewed and classified according to their quality of evidence using the grading system proposed by the Oxford Centre for Evidence-based Medicine. Seventeen randomized controlled trials (RCTs) were found for hepato-pancreatico-biliary surgery, none for upper GI tract, and 13 for lower GI tract surgery. If sufficient RCTs were identified, we performed a meta-analysis to characterize the drain effect using the random-effects model. There is evidence of level 1a that drains do not reduce complications after hepatic, colonic, or rectal resection with primary anastomosis and appendectomy for any stage of appendicitis. Drains were even harmful after hepatic resection in chronic liver disease and appendectomy. In the absence of RCTs, there is a consensus (evidence level 5) about the necessity of prophylactic drainage after esophageal resection and total gastrectomy due to the potential fatal outcome in case of anastomotic and gastric leakage. Many GI operations can be performed safely without prophylactic drainage. Drains should be omitted after hepatic, colonic, or rectal resection with primary anastomosis and appendectomy for any stage of appendicitis (recommendation grade A), whereas prophylactic drainage remains indicated after esophageal resection and total gastrectomy (recommendation grade D). For many other GI procedures, especially involving the upper GI tract, there is a further demand for well-designed RCTs to clarify the value of prophylactic drainage.


Recent advances in the use of neurotropic viruses for circuit analysis.
In the past decade, the literature describing how viruses can be used to define the synaptic architecture of the brain has increased dramatically. Early studies focused on determining the specificity of viral transport through synaptic connections. Analysis of the assembly and intracellular transport of viruses as well as of the role of the brain response to infection were central to this literature. With the growing acceptance that the transport of viruses is circuit-related, attention has shifted to application of the method to define the functional architecture of neural systems. The development of attenuated recombinant viruses that maintain neuroinvasiveness has been instrumental to the generation of increasingly powerful experimental approaches for the functional dissection of neural circuits. These approaches include the use of recombinant viruses that express unique reporters to address issues of axon collateralization in complex circuits, the use of green-fluorescent-protein-expressing recombinants to characterize the electrophysiological properties of projection-specific neurons in live slices of brain, and the exploitation of the Cre recombinase system for conditional replication of virus in phenotypically defined populations of neurons.


Evaluation of the pharmacokinetic equivalence and 54-week efficacy and safety of CT-P13 and innovator infliximab in Japanese patients with rheumatoid arthritis.
To demonstrate the pharmacokinetic equivalence of CT-P13 and its innovator infliximab (IFX) in Japanese patients with rheumatoid arthritis (RA), and to compare the efficacy and safety of these drugs, administered for 54 weeks. In a randomized, double-blind, parallel-group, multicenter study, 3 mg/kg of CT-P13 or IFX, in combination with methotrexate (MTX) (6-16 mg/week), was administered for 54 weeks to Japanese active RA patients with an inadequate response to MTX, to demonstrate the pharmacokinetic equivalence, based on the area under the curve (AUC(τ)) (weeks 6-14) and C(max) (week 6) of these drugs, and to compare their efficacy and safety. The CT-P13-to-IFX ratios (90% confidence intervals) of the geometric mean AUC(τ) and C(max) values in patients negative for antibodies to infliximab at week 14 were 111.62% (100.24-124.29%) and 104.09% (92.12-117.61%), respectively, demonstrating the pharmacokinetic equivalence of these drugs. In the full analysis set, CT-P13 and IFX showed comparable therapeutic effectiveness, as measured by the American College of Rheumatology, Disease Activity Score in 28 joints, the European League Against Rheumatism, and other efficacy criteria, at weeks 14 and 30. The incidence of adverse events was similar for these drugs. CT-P13 and IFX, administered at a dose of 3 mg/kg in combination with MTX to active RA patients, were pharmacokinetically equivalent and comparable in efficacy and safety.


Building models of animals from video.
This paper argues that tracking, object detection, and model building are all similar activities. We describe a fully automatic system that builds 2D articulated models known as pictorial structures from videos of animals. The learned model can be used to detect the animal in the original video--in this sense, the system can be viewed as a generalized tracker (one that is capable of modeling objects while tracking them). The learned model can be matched to a visual library; here, the system can be viewed as a video recognition algorithm. The learned model can also be used to detect the animal in novel images--in this case, the system can be seen as a method for learning models for object recognition. We find that we can significantly improve the pictorial structures by augmenting them with a discriminative texture model learned from a texture library. We develop a novel texture descriptor that outperforms the state-of-the-art for animal textures. We demonstrate the entire system on real video sequences of three different animals. We show that we can automatically track and identify the given animal. We use the learned models to recognize animals from two data sets; images taken by professional photographers from the Corel collection, and assorted images from the Web returned by Google. We demonstrate quite good performance on both data sets. Comparing our results with simple baselines, we show that, for the Google set, we can detect, localize, and recover part articulations from a collection demonstrably hard for object recognition.


An Integrated TCGA Pan-Cancer Clinical Data Resource to Drive High-Quality Survival Outcome Analytics.
For a decade, The Cancer Genome Atlas (TCGA) program collected clinicopathologic annotation data along with multi-platform molecular profiles of more than 11,000 human tumors across 33 different cancer types. TCGA clinical data contain key features representing the democratized nature of the data collection process. To ensure proper use of this large clinical dataset associated with genomic features, we developed a standardized dataset named the TCGA Pan-Cancer Clinical Data Resource (TCGA-CDR), which includes four major clinical outcome endpoints. In addition to detailing major challenges and statistical limitations encountered during the effort of integrating the acquired clinical data, we present a summary that includes endpoint usage recommendations for each cancer type. These TCGA-CDR findings appear to be consistent with cancer genomics studies independent of the TCGA effort and provide opportunities for investigating cancer biology using clinical correlates at an unprecedented scale.


The interaction of trh and dopaminergic mechanisms in the regulation of stimulated prolactin release in man.
The manner by which dopaminergic and TRH mechanisms interact to control PRL release is not known. Whilst dopamine receptor antagonists and TRH both release PRL, it is not known if the PRL released by these two mechanisms reflects similar aspects of physiological control, or if PRL responses to these mechanisms of release can be dissociated. We addressed this question by studying the PRL responses to maximal stimulatory dose of TRH and domperidone (a DA receptor antagonist), which were administered sequentially, simultaneously or separately on different occasions. Six normal volunteers undertook three sets of studies: (1) standard PRL stimulation tests to 400 micrograms TRH, 5 mg domperidone or simultaneous TRH/domperidone administration, (2) domperidone bolus-infusion study in which either 5 mg domperidone or 400 micrograms TRH was administered i.v. at 120 min during a 240 min infusion of domperidone (50 micrograms/min) which was preceded by a 5 mg i.v. bolus of the drug, and (3) TRH bolus-infusion study in which domperidone or TRH was administered i.v. at 120 min during a 240 min infusion of TRH (0.4 micrograms/min) which was preceded by a 400 micrograms i.v. bolus of the drug. In Study 1, simultaneous TRH/domperidone administration induced an incremental rise in PRL (5195 +/- 940 mIU/l) which was significantly greater (P less than 0.0005) than with either domperidone (3730 +/- 825 mIU/l) or TRH (1335 +/- 300 mIU/l) alone. In study 2, TRH administration at 120 min resulted in a significant rise (P less than 0.01) in PRL (delta PRL 960 +/- 232 mIU/l) whilst the second dose of domperidone did not, thus suggesting that the initial bolus and subsequent infusion had resulted in complete DA receptor blockade. In Study 3, domperidone administered at 120 min induced a marked rise in PRL (delta PRL 3609 +/- 963 mIU/l). In contrast, the corresponding TRH stimulus resulted in a small rise (delta PRL 142 +/- 32 mIU/l) suggesting that the PRL release induced by the initial bolus and subsequent infusion had been near maximal. Thus, TRH is able to induce significant PRL release in the presence of maximal DA receptor blockade, and domperidone, in the presence of maximal TRH stimulation, is also capable of inducing significant PRL release. These observations together with the ability of TRH/domperidone to induce a greater PRL response than either agent alone, suggest that each stimulus has a specific releasing action on a fraction of intracellular PRL which is not accessible to the other.(ABSTRACT TRUNCATED AT 400 WORDS)


Dietary sialic acid supplementation improves learning and memory in piglets.
Sialic acid, a key component of both human milk oligosaccharides and neural tissues, may be a conditional nutrient during periods of rapid brain growth. We tested the hypothesis that variations in the sialic acid content of a formula milk would influence early learning behavior and gene expression of enzymes involved in sialic acid metabolism in piglets. Piglets (n = 54) were allocated to 1 of 4 groups fed sow milk replacer supplemented with increasing amounts of sialic acid as casein glycomacropeptide for 35 d. Learning performance and memory were assessed with the use of easy and difficult visual cues in an 8-arm radial maze. Brain ganglioside and sialoprotein concentrations and mRNA expression of 2 learning-associated genes (ST8SIA4 and GNE) were measured. In both tests, the supplemented groups learned in significantly fewer trials than did the control group, with a dose-response relation for the difficult task (P = 0.018) but not the easy task. In the hippocampus, significant dose-response relations were observed between amount of sialic acid supplementation and mRNA levels of ST8SIA4 (P = 0.002) and GNE (P = 0.004), corresponding with proportionate increases in protein-bound sialic acid concentrations in the frontal cortex. Feeding a protein-bound source of sialic acid during early development enhanced learning and increased expression of 2 genes associated with learning in developing piglets. Sialic acid in mammalian milks could play a role in cognitive development.


A PCR-free fluorescence strategy for detecting telomerase activity via double amplification strategy.
As a universal tumor biomarker, research on the activity and inhibition of telomerase is of great importance for cancer diagnosis and therapy. Although the telomeric repeat amplification protocol (TRAP) has served as a powerful assay for detecting telomerase activity, its application has been significantly limited by amplification related errors and time-consuming procedure. To address the limitations of PCR-based protocol, a dual amplification fluorescence assay was developed for PCR-free detecting telomerase activity. Briefly, we designed an arch-structure DNA probe to specifically control strand displacement reaction and subsequent enzyme-aided amplification. Telomerase substrate (TS) primer was extended by telomerase to form long elongation products which contain several TTAGGG repeat units. So, one elongation product can release more than one trigger DNA (t-DNA) via strand displacement reaction to realize first amplification. Subsequently, t-DNA specifically opened molecular beacon (MB) to restore the fluorescence of MB. Meanwhile, t-DNA was recycled by the aid of nicking endonuclease to continuously open more and more MBs, leading to a second amplification. Owing to the double amplification strategy, the proposed method allowed the measurement of telomerase activity in crude cell extracts equivalent to 5 HeLa cells and 10 CCRF-CEM cells without PCR amplification. Besides, the influence of telomere-binding ligands on the telomerase activity demonstrated that the proposed method holds the potential to evaluate the inhibition efficiency of telomerase inhibitors.


Evolutionary impact assessment: accounting for evolutionary consequences of fishing in an ecosystem approach to fisheries management.
Managing fisheries resources to maintain healthy ecosystems is one of the main goals of the ecosystem approach to fisheries (EAF). While a number of international treaties call for the implementation of EAF, there are still gaps in the underlying methodology. One aspect that has received substantial scientific attention recently is fisheries-induced evolution (FIE). Increasing evidence indicates that intensive fishing has the potential to exert strong directional selection on life-history traits, behaviour, physiology, and morphology of exploited fish. Of particular concern is that reversing evolutionary responses to fishing can be much more difficult than reversing demographic or phenotypically plastic responses. Furthermore, like climate change, multiple agents cause FIE, with effects accumulating over time. Consequently, FIE may alter the utility derived from fish stocks, which in turn can modify the monetary value living aquatic resources provide to society. Quantifying and predicting the evolutionary effects of fishing is therefore important for both ecological and economic reasons. An important reason this is not happening is the lack of an appropriate assessment framework. We therefore describe the evolutionary impact assessment (EvoIA) as a structured approach for assessing the evolutionary consequences of fishing and evaluating the predicted evolutionary outcomes of alternative management options. EvoIA can contribute to EAF by clarifying how evolution may alter stock properties and ecological relations, support the precautionary approach to fisheries management by addressing a previously overlooked source of uncertainty and risk, and thus contribute to sustainable fisheries.


Measurement duration impacts variability but not impedance measured by the forced oscillation technique in healthy, asthma and COPD subjects.
The forced oscillation technique (FOT) is gaining clinical acceptance, facilitated by more commercial devices and clinical data. However, the effects of variations in testing protocols used in FOT data acquisition are unknown. We describe the effect of duration of data acquisition on FOT results in subjects with asthma, chronic obstructive pulmonary disease (COPD) and healthy controls. FOT data were acquired from 20 healthy, 22 asthmatic and 18 COPD subjects for 60 s in triplicate. The first 16, 30 and 60 s of each measurement were analysed to obtain total, inspiratory and expiratory resistance of respiratory system (Rrs) and respiratory system reactance (Xrs) at 5 and 19 Hz. With increasing duration, there was a decrease in total and expiratory Rrs for healthy controls, total and inspiratory Rrs for asthmatic subjects and magnitude of total and inspiratory Xrs for COPD subjects at 5 Hz. These decreases were small compared to the differences between clinical groups. Measuring for 16, 30 and 60 s provided ≥3 acceptable breaths in at least 90, 95 and 100% of subjects, respectively. The coefficient of variation for total Rrs and Xrs also decreased with duration. Similar results were found for Rrs and Xrs at 19 Hz. FOT results are statistically, but likely minimally, impacted by acquisition duration in healthy, asthmatic or COPD subjects.


Interpreting pathways to discover cancer driver genes with Moonlight.
Cancer driver gene alterations influence cancer development, occurring in oncogenes, tumor suppressors, and dual role genes. Discovering dual role cancer genes is difficult because of their elusive context-dependent behavior. We define oncogenic mediators as genes controlling biological processes. With them, we classify cancer driver genes, unveiling their roles in cancer mechanisms. To this end, we present Moonlight, a tool that incorporates multiple -omics data to identify critical cancer driver genes. With Moonlight, we analyze 8000+ tumor samples from 18 cancer types, discovering 3310 oncogenic mediators, 151 having dual roles. By incorporating additional data (amplification, mutation, DNA methylation, chromatin accessibility), we reveal 1000+ cancer driver genes, corroborating known molecular mechanisms. Additionally, we confirm critical cancer driver genes by analysing cell-line datasets. We discover inactivation of tumor suppressors in intron regions and that tissue type and subtype indicate dual role status. These findings help explain tumor heterogeneity and could guide therapeutic decisions.


Activity of preparations from Spilanthes oleracea, propolis, Nigella sativa, and black garlic on different microorganisms involved in oral diseases and on total human salivary bacteria: A pilot study.
Due to continuous rise in antibiotic resistance, there is a need for alternative treatment options to reduce the levels of oral pathogens for the maintenance of oral as well as overall health. The aim of this study was to evaluate the in vitro antibacterial potential of tinctures of Spilanthes oleracea and propolis, Nigella seed oil, and an ethanolic extract of black garlic on microorganisms involved in oral diseases. Both the minimum inhibitory concentration assay and the minimum bactericidal/fungicidal concentration assay were used in this study. Inhibition effects against total human salivary bacteria were also determined. Our results show that all of the preparations tested had potent antimicrobial activities. When measured 10 min after exposure, even low concentrations of the propolis tincture were found to have killed more than 99% of salivary bacteria, whereas Spilanthes tincture and black garlic extract killed more than 90% and Nigella seed oil more than 60% of the pathogens. This suggests that all preparations are promising candidates for the use in oral health care products and that all have the potential to control biofilm associated infections.


Defining the inherent stability of degenerative spondylolisthesis: a systematic review.
OBJECT A range of surgical options exists for the treatment of degenerative lumbar spondylolisthesis (DLS). The chosen technique inherently depends on the stability of the DLS. Despite a substantial body of literature dedicated to the outcome analysis of numerous DLS procedures, no consensus has been reached on defining or classifying the disorder with respect to stability or the role that instability should play in a treatment algorithm. The purpose of this study was to define grades of stability and to develop a guide for deciding on the optimal approach in surgically managing patients with DLS. METHODS The authors conducted a qualitative systematic review of clinical or biomechanical analyses evaluating the stability of and surgical outcomes for DLS for the period from 1990 to 2013. Research focused on nondegenerative forms of spondylolisthesis or spinal stenosis without associated DLS was excluded. The primary extracted results were clinical and radiographic parameters indicative of DLS instability. RESULTS The following preoperative parameters are predictors of stability in DLS: restabilization signs (disc height loss, osteophyte formation, vertebral endplate sclerosis, and ligament ossification), no disc angle change or less than 3 mm of translation on dynamic radiographs, and the absence of low-back pain. The validity and magnitude of each parameter's contribution can only be determined through appropriately powered prospective evaluation in the future. Identifying these parameters has allowed for the creation of a preliminary DLS instability classification (DSIC) scheme based on the preoperative assessment of DLS stability. CONCLUSIONS Spinal stability is an important factor to consider in the evaluation and treatment of patients with DLS. Qualitative assessment of the best available evidence revealed clinical and radiographic parameters for the creation of the DSIC, a decision aid to help surgeons develop a method of preoperative evaluation to better stratify DLS treatment options.


Alternative Lengthening of Telomeres in Primary Pancreatic Neuroendocrine Tumors Is Associated with Aggressive Clinical Behavior and Poor Survival.
Purpose: Alternative lengthening of telomeres (ALT), a telomerase-independent telomere maintenance mechanism, is strongly associated with ATRX and DAXX alterations and occurs frequently in pancreatic neuroendocrine tumors (PanNET).Experimental Design: In a Korean cohort of 269 surgically resected primary PanNETs and 19 sporadic microadenomas, ALT status and nuclear ATRX and DAXX protein expression were assessed and compared with clinicopathologic factors.Results: In PanNETs, ALT or loss of ATRX/DAXX nuclear expression was observed in 20.8% and 19.3%, respectively, whereas microadenomas were not altered. ALT-positive PanNETs displayed a significantly higher grade, size, and pT classification (all, P < 0.001). ALT also strongly correlated with lymphovascular (P < 0.001) and perineural invasion (P = 0.001) and the presence of lymph node (P < 0.001) and distant metastases (P = 0.002). Furthermore, patients with ALT-positive primary PanNETs had a shorter recurrence-free survival [HR = 3.38; 95% confidence interval (CI), 1.83-6.27; P < 0.001]. Interestingly, when limiting to patients with distant metastases, those with ALT-positive primary tumors had significantly better overall survival (HR = 0.23; 95% CI, 0.08-0.68; P = 0.008). Similarly, tumors with loss of ATRX/DAXX expression were significantly associated with ALT (P < 0.001), aggressive clinical behavior, and reduced recurrence-free survival (P < 0.001). However, similar to ALT, when limiting to patients with distant metastases, loss of ATRX/DAXX expression was associated with better overall survival (P = 0.003).Conclusions: Both primary ALT-positive and ATRX/DAXX-negative PanNETs are independently associated with aggressive clinicopathologic behavior and displayed reduced recurrence-free survival. In contrast, ALT activation and loss of ATRX/DAXX are both associated with better overall survival in patients with metastases. Therefore, these biomarkers may be used as prognostic markers depending on the context of the disease. Clin Cancer Res; 23(6); 1598-606. ©2016 AACR.


Patterns of Vasculature in Mouse Models of Lung Cancer Are Dependent on Location.
Preclinical studies of hypoxia are generally done using ectopic xenograft tumors, which behave differently from human tumors. Our previous findings have shown that subcutaneously implanted lung tumors exhibit more hypoxia than their orthotopic implanted or spontaneous K-ras-induced counterparts. We hypothesize that differences in hypoxia are due to site-specific differences in vascularity and perfusion. To compare the presence and functionality of vessels in these tumor models, we studied vascular perfusion in vivo in real time. Orthotopically implanted and spontaneous K-ras-induced lung tumors showed elevated perfusion, demonstrating vasculature functionality. Little contrast agent uptake was observed within the subcutaneously implanted tumors, indicating vascular dysfunction. These findings were corroborated at the microscopic level with Hoechst 33342 and Meca-32 staining. From these observations, we concluded that differences in hypoxia in experimental models is related to vessel perfusion. Thus, appropriate selection of preclinical lung tumor models is essential for the study of hypoxia, angiogenesis and therapies targeting these phenomena.


The limitations of using simple definitions of glucocorticoid exposure to predict fracture risk: A cohort study.
To evaluate the effects of different definitions of glucocorticoid (GC) exposure on the magnitude and pattern of fracture risk using the same dataset. Data from patients with rheumatoid arthritis (RA) were extracted from the Clinical Practice Research Datalink, a primary care database with electronic health records in the United Kingdom. Patients exposed to oral GCs were matched to up to two unexposed patients by age, gender and location. The first osteoporotic fracture was identified and adjusted and unadjusted cox proportional hazard ratios (HR) and 95% confidence intervals (CI) produced for fracture risk following GC therapy using different models of risk attribution. These include models demonstrating the effect of dose, duration and recency of GC exposure. There were 16,507 patients included. Exposed patients were older and had more comorbidities. GC therapy was associated with an increased risk of fracture, with the effect size influenced by risk attribution model. The risk of fracture decreased with less recent exposure from HR (95% CI) 1.66 (1.27, 2.16) during the first month of stopping GCs to 1.11 (0.79, 1.57) for between 1 and 3 months. The risk of fracture increased with current daily dose, HR 1.44 (1.17, 1.77) for 5-9.9 mg prednisolone equivalent dose (PEQ) to 3.02 (1.77, 5.15) for 15-19.9 mg PEQ. Risk of fracture increased with cumulative dose, a function of dose and duration, from HR 1.22 (1.03, 1.44) for <1 g to 1.83 (1.35, 2.48) for 7.5-10 g. GC exposure was associated with excess fracture risk, with effect size differing according to definition of exposure. This highlights the need to incorporate all exposure dimensions (dose, duration and recency) in these patient's fracture risk assessments.


Reading, laterality, and the brain: early contributions on reading disabilities by Sara S. Sparrow.
Although best known for work with children and adults with intellectual disabilities and autism spectrum disorders, training in speech pathology and a doctorate in clinical psychology and neuropsychology was the foundation for Sara Sparrow's long-term interest in reading disabilities. Her first papers were on dyslexia and laterality, and the maturational lag theory of developmental dyslexia proposed with Paul Satz, her mentor. The research program that emerged from this work had a wide impact on early neuropsychological models of reading disabilities. Although Sara went on to research focused on children with other developmental disabilities after she moved to Yale University, this initial research influenced her career- long interests in assessment, developmental models of disabilities, and early screening methods.


Antibody response of definitive hosts against antigens of two life stages of the neuropathogenic schistosome Trichobilharzia regenti.
The nasal avian schistosome Trichobilharzia regenti spends part of its intravertebrate period of life within the central nervous system. Migration of the parasites can be accompanied by neuromotor disorders or paralysis in natural definitive hosts (ducks) and even in laboratory mammals. Cercariae are also able to penetrate human skin and induce cercarial dermatitis. While the cellular and antibody responses against cercariae and migrating schistosomula have been investigated in mice, little is known about immune reactions in birds. This study first describes the dynamics of antibody response in infected ducks and identifies frequently recognized antigens that may serve as diagnostic markers of infection by T. regenti. Groups of 35 domestic ducks and 10 mallards were exposed to different doses of T. regenti cercariae. Sera were collected at predefined time intervals and tested by ELISA for the presence of specific anti-cercarial IgY and IgM. Antigens recognized by the antibodies were identified on Western blots of cercariae and schistosomula. The applicability in immunodiagnostics was statistically evaluated by expression of specificity and sensitivity values for individual antigens. In ELISA, the levels of anti-cercarial IgM peaked on day 15 pi. Increased production of IgY associated with the later phases of infection was observed in most individuals around 20 dpi and culminated 30 dpi. The time course of antibody response did not differ among experimental groups, variations were only observed in the levels of specific IgY which depended rather on the age of ducks at the time of infection than on the infectious dose. On Western blots, 40 cercarial and 7 schistosomular antigens were recognized by IgY from infected ducks. Among them, 4 cercarial antigens of 50, 47, 32 and 19 kDa provided the most sensitive and specific reactions. Antigens of cercariae and schistosomula elicited distinct antibody response in ducks, which correlated positively with the age of animals at the time of infection. Several antigens originating in cercariae and fewer in schistosomula were recognized by IgY with diverse sensitivity and specificity; only a few seemed to be common to both stages. Four of them were considered as the most promising candidates for immunodiagnostics.


On the suppression of vaccination dissent.
Dissenters from the dominant views about vaccination sometimes are subject to adverse actions, including abusive comment, threats, formal complaints,censorship, and de registration, a phenomenon that can be called suppression of dissent. Three types of cases are examined: scientists and physicians; a high-profile researcher; and a citizen campaigner. Comparing the methods used in these different types of cases provides a preliminary framework for understanding the dynamics of suppression in terms of vulnerabilities.


Pyoderma gangrenosum: proposed pathogenesis and current use of biologics with an emphasis on complement C5a inhibitor IFX-1.
Pyoderma gangrenosum (PG) is a rare neutrophilic dermatosis with no FDA-approved treatment. The complement pathway has received renewed attention because it is elevated in inflammatory cutaneous conditions such as hidradenitis suppurativa (HS) and psoriasis. IFX-1 is a complement C5a inhibitor which inhibits neutrophil activation, chemotaxis, and reduces inflammatory signaling and complement driven tissue damage in various diseases. The article discusses a proposed pathogenesis of PG, early clinical investigations of IFX-1 for the treatment of HS and PG, its potential as a treatment for PG, and those other biologics currently under investigation. Further studies should explore how patients with PG and other neutrophilic conditions may respond to complement inhibitors such as IFX-1. C5a blockade led to a reduction in inflammatory tunnels in HS, and alteration in neutrophil migration and activation supports the role of this pathway in the development of PG. The main challenges to the approval of IFX-1 are the identification of the optimal dose, duration, and stage-dependent factors in cutaneous inflammatory disorders. Further studies are required; however, complement inhibitors such as IFX-1 could find a place in clinical practice in years to come for severe, resistant PG that does not respond to conventional therapies.


Anatomy of the extensor mechanism in reference to quadriceps-sparing TKA.
The introduction of minimally invasive surgical techniques in total joint arthroplasty has ushered in a range of new terminology that often is unclear. One such term is quadriceps-sparing total knee arthroplasty (TKA). We examined 100 knees intraoperatively in 100 patients at the time of TKA, dissected 45 entire cadaveric leg specimens, and did high-resolution 3-Tesla magnetic resonance imaging scans on five normal knees to specifically determine: (1) the distal most insertion point of the vastus medialis obliquus; (2) the angle of insertion of the vastus medialis obliquus; and (3) the length of the vastus medialis obliquus tendon. The medial anatomy of the extensor mechanism was consistent. The inferior edge of the VMO inserted at or near the midpole of the patella in each case. The tendon inserted at 50 degrees (range, 46 degrees - 52 degrees). One hundred of the 150 patients (66%) had a tendon that measured 1.2 +/- 0.1 cm, whereas 50 of 150 patients (33%) had a substantially longer tendon (2.2 +/- 0.2 cm). Any medial arthrotomy that extends more proximal than the midpole of the patella detaches a portion of the quadriceps tendon. The term "quadriceps sparing" should not be applied to any surgical approach with a capsular incision that extends more proximal than the midpole of the patella.


Postoperative left ventricular apical ballooning: Transient Takotsubo cardiomyopathy following orthotopic liver transplantation.
Female, 51 FINAL DIAGNOSIS: Takotsubo cardiomyopathy Symptoms: - - Clinical Procedure: - Specialty: Cardiology • Transplantology. Rare disease. Left ventricular apical ballooning syndrome (LVAB), also known as Takotsubo cardiomyopathy, is a cardiac syndrome characterized by transient left ventricular dysfunction in the absence of obstructive atherosclerotic coronary artery disease. An episode of emotional stress, typically in female patients, is believed to precede and trigger the development of this syndrome. We report a case of Takotsubo cardiomyopathy that developed after orthotopic liver transplantation in a 51-year-old woman. On D2 (day 2) the patient had severe hemodynamic compromise. Echocardiography showed systolic dysfunction of the left ventricle (LV), with ejection fraction (EF) of 20% and anteroapical akinesis and ballooning of the apical 2/3 of the LV. Troponin T was elevated but other markers of myocardial necrosis were negative, as was coronary angiography. From D7 onward, there was an improvement in the hemodynamics in conjunction with a gradual increase of LV EF. The patient was dismissed from the hospital on D30 with signs of normal cardiac function and LV motion and EF of 50%. Liver function was also excellent. Every major operation, including liver transplantation, is associated with emotional stress for the patient. Therefore, it is necessary to consider Takotsubo cardiomyopathy in the differential diagnosis of heart failure developing early after LT, and clinicians should subsequently use adequate diagnostic and therapeutic measures.


The ethics of placebo-controlled trials in developing countries to prevent mother-to-child transmission of HIV.
Placebo-trials on HIV-infected pregnant women in developing countries like Thailand and Uganda have provoked recent controversy. Such experiments aim to find a treatment that will cut the rate of vertical transmission more efficiently than existing treatments like zidovudine. This scenario is first stated as generally as possible, before three ethical principles found in the Belmont Report, itself a sharpening of the Helsinki Declaration, are stated. These three principles are the Principle of Utility, the Principle of Autonomy and the Principle of Justice. These are taken as voices of moral imperative. But although each has intuitive appeal, it can be shown that there are possible scenarios in which they give conflicting prescriptions. To achieve consistency, one must be subordinate to the others. The voice of utility is taken as subordinate to those of justice and autonomy and it is shown that given plausible assumptions about the level of poverty and education in the developing country targeted, the experiment is ruled morally wrong in the name of both justice and autonomy. Moreover, it is argued that no justification can be found for the inclusion of a placebo group, when strictly defined. By contrast, a 'no-treatment' control arm might be justified, but only when the demands of autonomy are satisfied, demands that are more stringent than they might appear. A utilitarian defence of the experiment is examined, namely that the would-be participants are in a no-loss situation, and it is shown that this defence is seriously flawed. Finally, it is concluded that there is no justification for amending the Declaration of Helsinki.


EGR2 mutations define a new clinically aggressive subgroup of chronic lymphocytic leukemia.
Recurrent mutations within EGR2 were recently reported in advanced-stage chronic lymphocytic leukemia (CLL) patients and associated with a worse outcome. To study their prognostic impact, 2403 CLL patients were examined for mutations in the EGR2 hotspot region including a screening (n=1283) and two validation cohorts (UK CLL4 trial patients, n=366; CLL Research Consortium (CRC) patients, n=490). Targeted deep-sequencing of 27 known/postulated CLL driver genes was also performed in 38 EGR2-mutated patients to assess concurrent mutations. EGR2 mutations were detected in 91/2403 (3.8%) investigated cases, and associated with younger age at diagnosis, advanced clinical stage, high CD38 expression and unmutated IGHV genes. EGR2-mutated patients frequently carried ATM lesions (42%), TP53 aberrations (18%) and NOTCH1/FBXW7 mutations (16%). EGR2 mutations independently predicted shorter time-to-first-treatment (TTFT) and overall survival (OS) in the screening cohort; they were confirmed associated with reduced TTFT and OS in the CRC cohort and independently predicted short OS from randomization in the UK CLL4 cohort. A particularly dismal outcome was observed among EGR2-mutated patients who also carried TP53 aberrations. In summary, EGR2 mutations were independently associated with an unfavorable prognosis, comparable to CLL patients carrying TP53 aberrations, suggesting that EGR2-mutated patients represent a new patient subgroup with very poor outcome.


Positive selection of a duplicated UV-sensitive visual pigment coincides with wing pigment evolution in Heliconius butterflies.
The butterfly Heliconius erato can see from the UV to the red part of the light spectrum with color vision proven from 440 to 640 nm. Its eye is known to contain three visual pigments, rhodopsins, produced by an 11-cis-3-hydroxyretinal chromophore together with long wavelength (LWRh), blue (BRh) and UV (UVRh1) opsins. We now find that H. erato has a second UV opsin mRNA (UVRh2)-a previously undescribed duplication of this gene among Lepidoptera. To investigate its evolutionary origin, we screened eye cDNAs from 14 butterfly species in the subfamily Heliconiinae and found both copies only among Heliconius. Phylogeny-based tests of selection indicate positive selection of UVRh2 following duplication, and some of the positively selected sites correspond to vertebrate visual pigment spectral tuning residues. Epi-microspectrophotometry reveals two UV-absorbing rhodopsins in the H. erato eye with lambda(max) = 355 nm and 398 nm. Along with the additional UV opsin, Heliconius have also evolved 3-hydroxy-DL-kynurenine (3-OHK)-based yellow wing pigments not found in close relatives. Visual models of how butterflies perceive wing color variation indicate this has resulted in an expansion of the number of distinguishable yellow colors on Heliconius wings. Functional diversification of the UV-sensitive visual pigments may help explain why the yellow wing pigments of Heliconius are so colorful in the UV range compared to the yellow pigments of close relatives lacking the UV opsin duplicate.


CMS changes in reimbursement for HAIs: setting a research agenda.
The Centers for Medicare and Medicaid Services (CMS) promulgated regulations commencing October 1, 2008, which deny payment for selected conditions occurring during the hospital stay and are not present on admission. Three of the 10 hospital-acquired conditions covered by the new CMS policy involve healthcare-associated infections, which are a common, expensive, and often preventable cause of inpatient morbidity and mortality. To outline a research agenda on the impact of CMS's payment policy on the healthcare system and the prevention of healthcare-associated infections. An invitational day-long conference was convened in April 2009. Including the planning committee and speakers there were 41 conference participants who were national experts and senior researchers. Building upon a behavioral model and organizational theory and management research a conceptual framework was applied to organize the wide range of issues that arose. A broad array of research topics was identified. Thirty-two research agenda items were organized in the areas of incentives, environmental factors, organizational factors, clinical outcomes, staff outcomes, and financial outcomes. Methodological challenges are also discussed. This policy is a first significant step to move output-based inpatient funding to outcome-based funding, and this agenda is applicable to all hospital-acquired conditions. Studies beginning soon will have the best hope of capturing data for the years preceding the policy change, a key element in non-experimental research. The CMS payment policy offers an excellent opportunity to understand and influence the use of financial incentives for improving patient safety.


Copper complexes with dissymmetrically substituted bis(thiosemicarbazone) ligands as a basis for PET radiopharmaceuticals: control of redox potential and lipophilicity.
Copper(ii) bis(thiosemicarbazone) derivatives have been used extensively in positron emission tomography (PET) to image hypoxia and blood flow and to radiolabel cells for cell tracking. These applications depend on control of redox potentials and lipophilicity of the bis(thiosemicarbazone) complexes, which can be adjusted by altering peripheral ligand substituents. This paper reports the synthesis of a library of new dissymmetrically substituted bis(thiosemicarbazone) ligands by controlling the condensation reactions between dicarbonyl compounds and 4-substituted-3-thiosemicarbazides or using acetal protection. Copper complexes of the new ligands have been prepared by reaction with copper acetate or via transmetallation of the corresponding zinc complexes, which are convenient precursors for the rapid synthesis of radio-copper complexes. Well-defined structure-activity relationships linking ligand alkylation patterns with redox potential and lipophilicity of the complexes are reported.


Enhancing undergraduate community placements: a critical review of current literature.
In the face of the UK-wide policy shift to increased home care, inspiring and enabling the next generation of community nurses is more urgent than ever. The quality of the pre-registration practice learning experience is highly influential on career choices at the point of qualification. Given that 50% of learning by pre-registration students takes place in practice, mentors have a crucial role to play in preparing the next generation of nurses to work in the community. This article discusses the findings of a systematic and critical literature review of pre-registration placements that was funded by the Queen's Nursing Institute Scotland. The review found that students' experiences of learning in community settings are variable, and perceptions of students and mentors are misaligned in terms of what a quality placement should look like. Although there is no clear definition of what constitutes a community placement and there is some underuse of learning environments in areas such as general practice nursing, there are also a number of examples of new and imaginative placements. While these innovations provide 'whole experience' placements, they are currently lacking robust evaluation, despite their potential usefulness on a larger scale. Mentors have the opportunity to provide students with a range of learning opportunities that increase preparedness for working in the community, allowing final year students in particular greater influence over their learning experience. Students undertaking community practice learning, where they have a managed level of autonomy, are more likely to feel confident to take on community nursing roles.


Modulation of gut mucosal biofilms.
Non-digestible inulin-type fructans, such as oligofructose and high-molecular-weight inulin, have been shown to have the ability to alter the intestinal microbiota composition in such a way that members of the microbial community, generally considered as health-promoting, are stimulated. Bifidobacteria and lactobacilli are the most frequently targeted organisms. Less information exists on effects of inulin-type fructans on the composition, metabolism and health-related significance of bacteria at or near the mucosa surface or in the mucus layer forming mucosa-associated biofilms. Using rats inoculated with a human faecal flora as an experimental model we have found that inulin-type fructans in the diet modulated the gut microbiota by stimulation of mucosa-associated bifidobacteria as well as by partial reduction of pathogenic Salmonella enterica subsp. enterica serovar Typhimurium and thereby benefit health. In addition to changes in mucosal biofilms, inulin-type fructans also induced changes in the colonic mucosa stimulating proliferation in the crypts, increasing the release of mucins, and altering the profile of mucin components in the goblet cells and epithelial mucus layer. These results indicate that inulin-type fructans may stabilise the gut mucosal barrier. Dietary supplementation with these prebiotics could offer a new approach to supporting the barrier function of the mucosa.


Pyruvate administration reduces recurrent/moderate hypoglycemia-induced cortical neuron death in diabetic rats.
Recurrent/moderate (R/M) hypoglycemia is common in type 1 diabetes patients. Moderate hypoglycemia is not life-threatening, but if experienced recurrently it may present several clinical complications. Activated PARP-1 consumes cytosolic NAD, and because NAD is required for glycolysis, hypoglycemia-induced PARP-1 activation may render cells unable to use glucose even when glucose availability is restored. Pyruvate, however, can be metabolized in the absence of cytosolic NAD. We therefore hypothesized that pyruvate may be able to improve the outcome in diabetic rats subjected to insulin-induced R/M hypoglycemia by terminating hypoglycemia with glucose plus pyruvate, as compared with delivering just glucose alone. In an effort to mimic juvenile type 1 diabetes the experiments were conducted in one-month-old young rats that were rendered diabetic by streptozotocin (STZ, 50mg/kg, i.p.) injection. One week after STZ injection, rats were subjected to moderate hypoglycemia by insulin injection (10 U/kg, i.p.) without anesthesia for five consecutive days. Pyruvate (500 mg/kg) was given by intraperitoneal injection after each R/M hypoglycemia. Three hours after last R/M hypoglycemia, zinc accumulation was evaluated. Three days after R/M hypoglycemia, neuronal death, oxidative stress, microglial activation and GSH concentrations in the cerebral cortex were analyzed. Sparse neuronal death was observed in the cortex. Zinc accumulation, oxidative injury, microglial activation and GSH loss in the cortex after R/M hypoglycemia were all reduced by pyruvate injection. These findings suggest that when delivered alongside glucose, pyruvate may significantly improve the outcome after R/M hypoglycemia by circumventing a sustained impairment in neuronal glucose utilization resulting from PARP-1 activation.


Biological characterization of wild-bird-origin avian avulavirus 1 and efficacy of currently applied vaccines against potential infection in commercial poultry.
Newcastle disease virus (NDV), the type member of the species Avian avulavirus 1 (formerly known as avian paramyxovirus serotype 1), causes a highly contagious and economically important disease in a myriad of avian species around the globe. While extensive vaccination programs have been implemented in ND-endemic countries, the disease is continuously spreading in commercial, backyard, and wild captive poultry. In order to investigate the evolution of the virus and assess the efficiency of the vaccine regimens that are currently being applied in commercial poultry, four wild-bird-origin NDV strains were characterized biologically, based on mean death time and intracerebral pathogenicity index, and genetically, based on the cleavage motif (112RRQKRF117) in the fusion (F) protein. Based on these features, all of the isolates were characterized as velogenic strains of NDV. Phylogenetic analysis based on the complete genome sequence revealed clustering of these isolates within class II, genotype VII. This class of NDV remains the predominant genotype in the Egyptian poultry industry, as well as in those of many Asian and African countries. To investigate the potential of these wild-bird-origin NDV isolates to cause infection in domesticated poultry and to assess the efficacy of currently available vaccines for protection of commercial poultry, an extensive animal challenge experiment was performed. Cumulative clinicopathological and immunological investigations of virus-challenged chickens indicate that these isolates can potentially be transmitted between chicken and cause systemic infections, and the currently applied vaccines are unable to prevent clinical disease and virus shedding. Taken together, the data represent a comprehensive evaluation of the ability of Egyptian wild-bird-origin NDV strains to cause infection in commercial poultry and highlights the need for a continuous and large-scale surveillance as well as revised vaccine approaches. These integrated and multifaceted strategies would be crucial in any efforts to control and eradicate the disease globally.


Sphingosine-1-phosphate receptor-independent lung endothelial cell barrier disruption induced by FTY720 regioisomers.
Vascular permeability is a hallmark of acute respiratory distress syndrome (ARDS) and ventilator-induced lung injury pathobiology; however, the mechanisms underlying this vascular dysregulation remain unclear, thereby impairing the development of desperately needed effective therapeutics. We have shown that sphingosine-1-phosphate (S1P) and 2-amino-2-(2-[4-octylphenyl]ethyl)-1,3-propanediol (FTY720) analogues are useful tools for exploring vascular barrier regulation mechanisms. To experimentally define the effects of FTY720 regioisomers on lung endothelial cell barrier regulation. Specific barrier-regulatory receptor and kinase inhibitors were utilized to probe signaling mechanisms involved in FTY720 regioisomer-mediated human lung endothelial cell barrier responses (trans-endothelial electrical resistance, TER). Docking simulations with the S1P1 receptor were performed to further evaluate FTY720 regioisomer signaling. FTY720 regioisomers produced potent endothelial cell barrier disruption reflected by declines in TER alterations. Pharmacologic inhibition of Gi-coupled S1P receptors (S1P1, S1P2, S1P3) failed to alter FTY720 regioisomer-mediated barrier disruption; findings that were corroborated by docking simulations demonstrating FTY720 regiosomers were repelled from S1P1 docking, in contrast to strong S1P1 binding elicited by S1P. Inhibition of either the barrier-disrupting PAR-1 receptor, the VEGF receptor, Rho-kinase, MAPK, NFkB, or PI3K failed to alter FTY720 regioisomer-induced endothelial cell barrier disruption. While FTY720 regioisomers significantly increased protein phosphatase 2 (PP2A) activity, PP2A inhibitors failed to alter FTY720 regioisomer-induced endothelial cell barrier disruption. Together, these results imply a vexing model of pulmonary vascular barrier dysregulation in response to FTY720-related compounds and highlight the need for further insights into mechanisms of vascular integrity required to promote the development of novel therapeutic tools to prevent or reverse the pulmonary vascular leak central to ARDS outcomes.


Preliminary Investigation of the Perspectives of Parents of Children With Cerebral Palsy on the Supports, Challenges, and Realities of Integrating Augmentative and Alternative Communication Into Everyday Life.
Purpose To ensure long-term adoption and use of augmentative and alternative communication (AAC) technologies, they must be designed to support children to participate within everyday activities and routines that are prioritized by families. The aim of this study was to gain parent perspectives on how AAC technologies were integrated into everyday life. Method Nine parents of children with cerebral palsy who used AAC technologies participated in semistructured interviews to provide their perspectives on how AAC technologies were integrated into the functional contexts of everyday life. Results Five major themes emerged from the discussions: (a) integrating AAC into life, (b) AAC technologies, (c) child needs and skills, (d) parent responsibilities and priorities, and (e) AAC process and decision making. Children were able to use AAC technologies within a variety of everyday contexts with various partners; however, challenges included access to technologies within care routines and outdoor activities as well as partners who lacked knowledge regarding operational competencies and effective interaction strategies. To integrate AAC technologies into life, parents prioritized technology features including ease of programming, improved physical design, features to enhance efficiency and ease of access, and availability of multiple functions and features (e.g., games and leisure activities, environmental controls). Conclusion AAC manufacturers and mainstream technology developers should work to ensure that technologies are responsive to the supports, limitations, and ideal features identified by parents. Future research should seek input from a larger group of stakeholders and use longitudinal methods to examine perceptions of AAC technologies over time. Supplemental Material https://doi.org/10.23641/asha.11625543.


Digital speckle-displacement measurement using a complex spectrum method.
An alternative approach to fully automatic speckle-displacement measurement is described. Two speckle patterns of a specimen, one before and one after deformation, are captured by a CCD camera and registered by a frame grabber. Two series of small subimages are obtained by segmenting the two speckle patterns. The corresponding subimage pairs extracted from both series are analyzed pointwise. The interrogation of each subimage pair involves a two-step fast-Fourier transform. While the first-step fast-Fourier transform achieves a complex spectrum characterized by the local displacement information, the second-step one generates a signal peak in the second spectral domain that resolves the local displacement vector. A rough estimate of the displacement vector is achieved by detecting the maximum pixel of the discrete spectrum. A more accurate determination is attained by a subpixel-maximum determination through a biparabolic fitting near the signal peak. The u- and v-displacement fields are deduced by analyzing all subimage pairs. A large rigid-body displacement can be overcome by introducing an artificial rigid shift of the two speckle patterns toward each other before the numerical process. The technique retains all the advantages of optical speckle photography and provides an extended range of measurement. Dynamic incremental deformations can be inspected by registering more speckle patterns at many consecutive deformation stages by using a high-speed CCD camera. The system was applied successfully to the study of crack-tip deformation fields.


Onlay bone graft maintenance using guided bone regeneration, platelet rich plasma, and their combination.
Onlay bone grafts have a bad reputation of resorption with loss of contour and volume. Rigid fixation reduces the incidence of resorption but does not prevent it. Literature shows reduction of resorption by applying guided bone regeneration (GBR) barriers and platelet-rich plasma (PRP). Investigating the effect of combining them together to reduce resorption was the aim of this study. This study included 4 groups: control group, GBR group, PRP group, and GBR + PRP group. Twenty rabbits were used (40 mandibular halves). Onlay bone grafts were fixed by titanium miniscrews in all groups. Computed tomography scans of harvested mandibles after euthanasia allowed calculations of bone graft volume and density. Onlay bone graft volumes in all experimental groups were significantly higher than in the control group. Volume maintenance in the GBR group was significantly higher than in the PRP group. There was no significant difference in the volume of onlay bone grafts between the group of combined GBR + PRP and GBR alone. It was concluded that, to maintain the volume of onlay bone grafts, either GBR or PRP can be added. Combining them did not add any advantage over the GBR alone.


Classical and molecular cytogenetic characterization of allochthonous European bitterling Rhodeus amarus (Cyprinidae, Acheilognathinae) from Northern Italy.
A cytogenetical study was carried out on 34 specimens of the European bitterling Rhodeus amarus (Teleostei: Cyprinidae, Acheilognathinae) from four rivers of the Venice district (NE Italy). This allochthonous fish species was accidentally introduced in the North-East of Italy about 20 years ago and is now rapidly spreading all over the rivers of the Northern part of the country. All the studied specimens are characterised by the same karyotype (2n = 48: 8M + 20SM + 20ST), i.e., the typical one of the native populations of the species. However, a polymorphism in the number of NOR bearing chromosomes has been found. In fact, in addition to the main species-specific NORs, on the short arms of chromosome pair 7, two to five additional 18S rDNA sites have been revealed by FISH in different specimens. Sequential staining with silver nitrate, chromomycin A(3) and DAPI revealed that most of the additional sites are inactive and CMA(3)-positive. Data herein reported confirm that in spite of an overall morphological karyological conservativeness, significant differences for the finer cytogenetic features can be found within the Acheilognathinae with the 2n = 48 and NF = 76 karyotype.


Oncofertility-An emerging discipline rather than a special consideration.
Originally absent from the oncologist's consult, then placed in a 'quality of life' rubric, oncofertility should now be an essential part of a comprehensive cancer treatment plan in patients of reproductive age, including adolescents and young adults (AYAs). Oncofertility encompasses the endocrine health of the patient, as well as fertility management options. Thus, pubertal transitions in males and females, bone health, and menstrual health are all part of this discipline, enabling practitioners to work in interdisciplinary teams to solve problems in reproductive health. This review provides a summary of the essential considerations required for the assessement of reproductive risk and choice of fertility preservation options as well as considerations for developing oncofertility services for AYAs.


Relationship between oxidative stress and inflammatory cytokines in diabetic nephropathy.
The prevalence of diabetes has dramatically increased worldwide due to the vast increase in the obesity rate. Diabetic nephropathy is one of the major complications of type 1 and type 2 diabetes and it is currently the leading cause of end-stage renal disease. Hyperglycemia is the driving force for the development of diabetic nephropathy. It is well known that hyperglycemia increases the production of free radicals resulting in oxidative stress. While increases in oxidative stress have been shown to contribute to the development and progression of diabetic nephropathy, the mechanisms by which this occurs are still being investigated. Historically, diabetes was not thought to be an immune disease; however, there is increasing evidence supporting a role for inflammation in type 1 and type 2 diabetes. Inflammatory cells, cytokines, and profibrotic growth factors including transforming growth factor-β (TGF-β), monocyte chemoattractant protein-1 (MCP-1), connective tissue growth factor (CTGF), tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-18 (IL-18), and cell adhesion molecules (CAMs) have all been implicated in the pathogenesis of diabetic nephropathy via increased vascular inflammation and fibrosis. The stimulus for the increase in inflammation in diabetes is still under investigation; however, reactive oxygen species are a primary candidate. Thus, targeting oxidative stress-inflammatory cytokine signaling could improve therapeutic options for diabetic nephropathy. The current review will focus on understanding the relationship between oxidative stress and inflammatory cytokines in diabetic nephropathy to help elucidate the question of which comes first in the progression of diabetic nephropathy, oxidative stress, or inflammation.


Long-term exposure to increasing temperature can offset predicted losses in marine food quality (fatty acids) caused by ocean warming.
Marine phytoplankton produce essential fatty acids (FA), which are key component of a healthy diet in humans and marine food webs. Increased temperatures can reduce lipid and FA content in phytoplankton; thus, ocean warming poses a risk for the global production of these essential FA. However, responses to warming may differ between phytoplankton species especially after long-term exposure because phenotypic plasticity, de novo mutations, or genetic evolution may occur. Here, we examine the content of FA and lipids in phytoplankton following long-term selection (~2 years) to warming conditions (+4°C), and we observe that FA and lipids content were partly or entirely recovered following long-term exposure to warming conditions. Furthermore, this observed long-term response also offset the predicted losses of some essential polyunsaturated fatty acids (PUFA) in three of the four species tested. Our study suggests that long-term exposure of phytoplankton to warming may help to maintain marine food quality in a moderately warming ocean. The responses of FA to increasing temperatures may vary among species, and the level of this idiosyncrasy remains to be further studied.


Metal-Support Interactions of Platinum Nanoparticles Decorated N-Doped Carbon Nanofibers for the Oxygen Reduction Reaction.
N-doped carbon materials are discussed as catalyst supports for the electrochemical oxygen reduction reaction (ORR) in fuel cells. This work deals with the preparation of Pt nanoparticles (NPs) supported on N-doped carbon nanofibers (N-CNF) from a polyaniline nanofiber (PANI NF) precursor, and investigates the ORR activity of the produced materials. Initially, Pt NPs are deposited on PANI NFs. The PANI NF precursors are characterized by near-edge X-ray absorption fine structure (NEXAFS) and transmission electron microscopy (TEM) measurements. It is shown, that in the PANI NF precursor materials electrons from the Pt are being transferred toward the π-conjugated systems of the aromatic ring. This strong interaction of Pt atoms with PANI explains the high dispersion of Pt NPs on the PANI NF. Subsequently, the PANI NF precursors are carbonized at different heat-treatment conditions resulting in structurally different N-CNFs which are characterized by NEXAFS, X-ray photoelectron spectroscopy (XPS) ,and TEM measurements. It is shown that an interaction between N-groups and Pt NPs exists in all investigated N-CNFs. However, the N-CNFs differ in the composition of the N-species and the dispersion of the Pt NPs. A small mean Pt NP size with a narrow size distribution is attributed to the presence of pyrdinic N-groups in the N-CNFs, whereas, for the N-CNFs with mainly graphitic and pyrrolic N-groups, an increase in the average Pt NP size with a broad size distribution is found. The ORR activity in alkaline media investigated by Koutecky-Levich analysis of rotating disk electrode measurements showed a largely enhanced ORR activity in comparison to a conventional Pt/C catalyst.


Salivary hormone concentrations and technical-tactical performance indicators in beach volleyball: Preliminary evidence.
The present study aimed to investigate (i) differences in salivary testosterone and cortisol concentrations before, during, and after simulated beach volleyball match, depending on match outcome (winning vs. losing); (ii) the relationship between technical-tactical performance indicators in beach volleyball and salivary hormonal concentrations (i.e., testosterone, cortisol). We hypothesized (i) salivary testosterone concentrations would be greater in winners and salivary cortisol would be lower; (ii) testosterone would associate with positive technical-tactical performance and cortisol would associate with negative technical-tactical performance. Sixteen athletes participated in the study and were grouped according to the result of a simulated game (winners: n = 8; losers: n = 8). Salivary hormone concentration of testosterone and cortisol were measured by enzyme-linked immunosorbent assay (pre-match, post first set, and post-match), and the coefficient of performance and efficiency were used as technical-tactical performance indicators. Regarding testosterone, there was a large effect size for match outcome after the first set (i.e., Winner vs. Losers) and a moderate effect size for the time in winners (pre-match vs. post-match). Regarding cortisol, there was a moderate effect size of time in losers only (pre-match vs. post-match). Moreover, cortisol pre-match was negatively correlated with the offensive performance (attack performance coefficient: r = -0.541; p = 0.030; attack efficiency: r = -0.568; p = 0.022). In conclusion, the effect of match outcome on testosterone and cortisol levels was moderate in winners and losers, respectively. Moreover, resting cortisol concentration appears to be related to a diminished attack technical-tactical performance. However, larger confirmatory studies are required to confirm these data to corroborate winning increases testosterone levels and/or reduces cortisol in a sporting setting.


Full down-conversion of amber-emitting phosphor-converted light-emitting diodes with powder phosphors and a long-wave pass filter.
This paper reports the possibility of a facile optical structure to realize a highly efficient monochromatic amber-emitting light-emitting diode (LED) using a powder-based phosphor-converted LED combined with a long-wave pass filter (LWPF). The capping of a blue-reflecting and amber-passing LWPF enhances both the amber emission from the silicate amber phosphor layer and the color purity due to the blocking and recycling of the pumping blue light from the InGaN LED. The enhancement of the luminous efficacy of the amber pc-LED with a LWPF (phosphor concentration 20 wt%, 39.4 lm/W) is 34% over that of an amber pc-LED without a LWPF (phosphor concentration 55 wt%, 29.4 lm/W) at 100 mA and a high color purity (>96%) with Commission International d'Eclairage (CIE) color coordinates of x=0.57 and y=0.42.


Evaluation of a stage-specific proteolytic enzyme of Schistosoma mansoni as a marker of exposure.
Cercarial elastase (CE) is one of the first proteins released in the host by invading schistosome cercariae. An enzyme-linked immunosorbent assay (ELISA)-formatted immunoassay has been developed to detect antibodies to the stage-specific CE antigen of Schistosoma mansoni as marker of exposure. We have evaluated this test system as an epidemiologic tool, using well-characterized sera collected from S. mansoni- and S. haematobium-infected subjects residing in endemic areas and from control subjects living in nonendemic areas in Egypt. Urine, stool specimens, and blood samples were collected from a sample of 272 endemic subjects randomly selected to represent different age groups in the range of 2-20 years of age. Of 47 S. mansoni-infected subjects, 41 (87.2%) had anti-CE IgG antibodies. Of 52 S. haematobium-infected cases, 38 (73.0%) had IgM antibodies to CE and 43 (82.7%) had IgG antibodies to CE. Of 173 egg-negative people in the endemic area, 84 (48.6%) were IgM positive and 99 (57.2%) were IgG positive. The mean IgM and IgG antibody levels were similar in the infected groups but were significantly lower in the egg-negative group (P = 0.001). All sera from young children (2-3 years of age) were uniformly ELISA negative. The prevalence of IgM and IgG antibodies to CE in children less than six years of age were significantly lower than in other age groups. There was no significant difference in prevalence rates of IgM and IgG anti-CE antibodies between subjects having other parasites present in the endemic area (Ascaris lumbricoides, Entrobius vermicularis, Hymenolepis nana, H. diminuta, Trichostrongylus spp., and Entamoeba histolytica) and those without any parasitic infection. All nonendemic sera (58), including those with other helminth infections, were uniformly ELISA negative for antibodies to CE. These findings suggest that antibodies to elastase indicate exposure, but not necessarily active schistosome infection.


The role of "disaggregation" in optical probe development.
"Aggregation-caused signal change" is a well-established mechanism by now and has been widely used as the basis for optical probe and sensor development. Compared to aggregation, its reverse process, disaggregation, has received much less attention and is not properly discussed in the literature so far. With the less established paradigm or mechanism, although some of the reported sensors and probes seem to work through disaggregation phenomena, the proper interpretation of the results and applying the concept to novel probe development is seriously hampered. The process from aggregation to disaggregation generally causes a recovery or enhancement of fluorescence signals, and thus provides an interesting new path to design "turn-on" probes. This tutorial review will provide the balanced comparison between aggregation and disaggregation mechanism, and focuses on the less explored advantages of "disaggregation" as a novel sensing mechanism and its recent applications in probe development.


Dynamics Rationalize Proteolytic Susceptibility of the Major Birch Pollen Allergen Bet v 1.
Proteolytic susceptibility during endolysosomal degradation is decisive for allergic sensitization. In the major birch pollen allergen Bet v 1 most protease cleavage sites are located within its secondary structure elements, which are inherently inaccessible to proteases. The allergen thus must unfold locally, exposing the cleavage sites to become susceptible to proteolysis. Hence, allergen cleavage rates are presumed to be linked to their fold stability, i.e., unfolding probability. Yet, these locally unfolded structures have neither been captured in experiment nor simulation due to limitations in resolution and sampling time, respectively. Here, we perform classic and enhanced molecular dynamics (MD) simulations to quantify fold dynamics on extended timescales of Bet v 1a and two variants with higher and lower cleavage rates. Already at the nanosecond-timescale we observe a significantly higher flexibility for the destabilized variant compared to Bet v 1a and the proteolytically stabilized mutant. Estimating the thermodynamics and kinetics of local unfolding around an initial cleavage site, we find that the Bet v 1 variant with the highest cleavage rate also shows the highest probability for local unfolding. For the stabilized mutant on the other hand we only find minimal unfolding probability. These results strengthen the link between the conformational dynamics of allergen proteins and their stability during endolysosomal degradation. The presented approach further allows atomistic insights in the conformational ensemble of allergen proteins and provides probability estimates below experimental detection limits.


Attenuation of oxidant/antioxidant imbalance during treatment of exacerbations of chronic obstructive pulmonary disease.
An oxidant/antioxidant imbalance is thought to occur in patients with chronic obstructive pulmonary disease (COPD). It has recently been shown that during exacerbations of COPD the antioxidant capacity and protein sulfhydryls of plasma are lower and the levels of products of lipid peroxidation are higher than in age matched healthy subjects. The aims of this study were to confirm these data and to measure the time course of these changes. The plasma Trolox equivalent antioxidant capacity (TEAC), protein sulfhydryls, and products of lipid peroxidation were measured throughout the course of treatment in 13 patients who presented with an acute exacerbation of COPD. TEAC values (mmol/l) were low on admission (mean 0.67, 95% confidence interval (CI) 0.32 to 0.88; p < 0.05) and had increased by discharge (0.98, 95% CI 0.88 to 1.2; p < 0.05) but still remained lower than in healthy subjects (1.33, 95% CI 1.11 to 1.65). There was also restoration of plasma protein sulfhydryl levels (mmol/l) from admission (0.32, 95% CI 0.20 to 0.43) to discharge (0.49, 95% CI 0.42 to 0.62, p < 0.001) to levels similar to those in healthy subjects (0.52, 95% CI 0.43 to 0.65). Products of lipid peroxidation, measured as thiobarbituric acid-malondialdehyde adducts (mumol/l), were significantly higher (2.08, 95% CI 1.8 to 2.5) than in control subjects (1.3, 95% CI 0.85 to 1.32; p < 0.01) and returned to normal values by the time of discharge (1.2, 95% CI 0.88 to 1.29). These data confirm the presence of a profound oxidant/antioxidant imbalance in the blood of patients with acute exacerbations of COPD which returns towards normal values during the course of treatment.


Using knockout mutants to reveal the growth costs of defensive traits.
We used a selection of Arabidopsis thaliana mutants with knockouts in defence genes to demonstrate growth costs of trichome development and glucosinolate production. Four of the seven defence mutants had significantly higher size-standardized growth rates (SGRs) than the wild-type in early life, although this benefit declined as plants grew larger. SGR is known to be a good predictor of success under high-density conditions, and we confirmed that mutants with higher growth rates had a large advantage when grown in competition. Despite the lack of differences in flowering-time genes, the mutants differed in flowering time, a trait that strongly correlated with early growth rate. Aphid herbivory decreased plant growth rate and increased flowering time, and aphid population growth rate was closely coupled to the growth rate of the host plant. Small differences in early SGR thus had cascading effects on both flowering time and herbivore populations.


Operational use of neem oil as an alternative anopheline larvicide. Part A: Laboratory and field efficacy.
We conducted a study to determine the laboratory and field efficacy of neem oil towards anopheline larvae. No difference in LC50 was observed between laboratory and field strains for temephos, chlorpyriphos-methyl/fenitrothion and neem oil. No difference in susceptibility was found after 3 months of application every 2 weeks. Water treated with a single application of traditional larvicides was free of larvae after 4 weeks; neem oil-treated water, however, was free after 2 weeks but not at 4 weeks. Application of chlorpyriphos-methyl/fenitrothion and neem oil every 2 weeks for 7 rounds resulted in dramatic reduction in larval density with no statistically significant differences. An adult survey after larviciding also showed no significant difference. The efficacy of crude neem oil appears to be below that of conventional larvicides.


Demand and capacity in an ADHD team: reducing the wait times for an ADHD assessment to 12 weeks.
Attention deficit hyperactivity disorder (ADHD) is a behavioural disorder characterised by the core symptoms of hyperactivity, impulsivity and inattention. ADHD is thought to affect about 3%-9% of school-age children and young people in the UK. With increased awareness and early identification of ADHD, and the long-term impact of the condition, there is a growing demand for ADHD services for both assessment and treatment of children and young people with the condition. Demand and capacity modelling carried out in October 2017 identified the ADHD pathway team in City and Hackney Child and Adolescent Mental Health Service (CAMHS) were working at 127% utilisation, indicating a mismatch between capacity and demand. A quality improvement (QI) project was implemented to improve efficiency and effectiveness of processes within the team and to support the increasing demand within the limited capacity and resource. The aim of the project was to reduce the average length of time from initial referral to CAMHS to 'ADHD assessment feedback' to 12 weeks by September 2018, which is in line with trust-level targets. The team followed the model for improvement and guidance from East London Foundation Trust (ELFT) QI Microsite to structure the project. They used a variety of tools to develop a theory of change, and used Plan-Do-Study-Act cycles to test change ideas. Overall wait times have reduced from 28 weeks to below our target of 12 weeks. Data examining the entry point to the ADHD pathway to completion of the ADHD assessment and feedback reduced from an average of 87 days, to an average of 18 days.The diagnostic rate has increased from 62% to 78% (due to more appropriate screening and referrals). The QI approach was systematic and supported the development of more efficient systems; reducing wait times and increasing capacity to manage the demand. Team engagement in 'change', by embedding QI into fortnightly team meetings, has resulted in collective ownership and responsibility across team members. A monitoring system is supporting the sustainability and maintenance of improvement.


Integrating geo-referenced multiscale and multidisciplinary data for the management of biodiversity in livestock genetic resources.
In livestock genetic resource conservation, decision making about conservation priorities is based on the simultaneous analysis of several different criteria that may contribute to long-term sustainable breeding conditions, such as genetic and demographic characteristics, environmental conditions, and role of the breed in the local or regional economy. Here we address methods to integrate different data sets and highlight problems related to interdisciplinary comparisons. Data integration is based on the use of geographic coordinates and Geographic Information Systems (GIS). In addition to technical problems related to projection systems, GIS have to face the challenging issue of the non homogeneous scale of their data sets. We give examples of the successful use of GIS for data integration and examine the risk of obtaining biased results when integrating datasets that have been captured at different scales.


Efficacy of fluticasone on cough: a randomised controlled trial.
Cough may be the consequence of bronchial hyperresponsiveness (BHR) and inflammation. This study was designed to investigate the short-term effects of an inhaled steroid (fluticasone propionate (FP)) on cough, and to determine the effects of smoking, BHR, allergy and forced expiratory volume in one second (FEV1) on the efficacy of FP. In a community-based primary healthcare centre, 135 previously healthy adults suffering from cough for > or =2 weeks were enrolled in a randomised, double-blind, placebo-controlled trial of inhaled FP 500 microg b.i.d. for 2 weeks. Participants completed daily diary cards of lower respiratory tract symptoms. The primary outcome measure was the decrease in mean total daily cough score (0-6) during the second week of treatment. In the FP group, the cough score decreased from 3.8 at baseline to mean+/-SEM 1.4+/-0.2 during the second week. In the placebo group, this decrease was from 3.8 to 1.9+/-0.1 and was statistically significantly less. A favourable effect of FP was only detectable in nonsmokers, in whom the score was 0.9 points lower compared with placebo. The clinical relevance of this finding has to be established further. Allergy, FEV1 and BHR at baseline did not affect the efficacy of FP. In conclusion, anti-inflammatory treatment with the inhaled steroid fluticasone propionate reduces cough in otherwise healthy adults who do not smoke.


Structure-functional intimacies of transient receptor potential channels.
Although a unifying characteristic common to all transient receptor potential (TRP) channel functions remains elusive, they could be described as tetramers formed by subunits with six transmembrane domains and containing cation-selective pores, which in several cases show high calcium permeability. TRP channels constitute a large superfamily of ion channels, and can be grouped into seven subfamilies based on their amino acid sequence homology: the canonical or classic TRPs, the vanilloid receptor TRPs, the melastatin or long TRPs, ankyrin (whose only member is the transmembrane protein 1 [TRPA1]), TRPN after the nonmechanoreceptor potential C (nonpC), and the more distant cousins, the polycystins and mucolipins. Because of their role as cellular sensors, polymodal activation and gating properties, many TRP channels are activated by a variety of different stimuli and function as signal integrators. Thus, how TRP channels function and how function relates to given structural determinants contained in the channel-forming protein has attracted the attention of biophysicists as well as molecular and cell biologists. The main purpose of this review is to summarize our present knowledge on the structure of channels of the TRP ion channel family. In the absence of crystal structure information for a complete TRP channel, we will describe important protein domains present in TRP channels, structure-function mutagenesis studies, the few crystal structures available for some TRP channel modules, and the recent determination of some TRP channel structures using electron microscopy.


A comparison of the Ranging behaviour and habitat use of the Ethiopian hedgehog (Paraechinus aethiopicus) in Qatar with hedgehog taxa from temperate environments.
We investigated seasonal changes in the ecology and behaviour of the Ethiopian hedgehog (Paraechinus aethiopicus) in Qatar, particularly in respect to differences in behaviour between hedgehogs living in arid environments and hedgehogs in temperate mesic environments. These comparisons will allow us to explore behavioural adaptations to different environments across hedgehog taxa. We radio-tracked 30 hedgehogs in Qatar over two years, and measured home range size, habitat preference, travel speed, activity and body mass. Whilst we found no difference in body mass between males and females, male home range size was over twice as large as that for females. Unlike hedgehogs in Europe, males maintained large home ranges during the non-breeding season. This behaviour may be sustained by the low cost of maintaining a large home range; males travelled less far per hour during the non-breeding season. Habitat use was non-random; arid areas with human influence, including rubbish dumping sites, was the most selected habitat type compared with its availability. Dense scrub and/or trees was the most selected habitat for nesting. This study gives us greater understanding as to how hedgehog taxa are adapted to their environment and therefore how they may be conserved, for example, the recent increase of "lower" level human activities, including irrigated farms and food waste, in harsh arid environments may have influenced the space use by Ethiopian hedgehogs.


The upper-airway microbiota and loss of asthma control among asthmatic children.
The airway microbiome has an important role in asthma pathophysiology. However, little is known on the relationships between the airway microbiome of asthmatic children, loss of asthma control, and severe exacerbations. Here we report that the microbiota's dynamic patterns and compositions are related to asthma exacerbations. We collected nasal blow samples (n = 319) longitudinally during a clinical trial at 2 time-points within one year: randomization when asthma is under control, and at time of early loss of asthma control (yellow zone (YZ)). We report that participants whose microbiota was dominated by the commensal Corynebacterium + Dolosigranulum cluster at RD experience the lowest rates of YZs (p = 0.005) and have longer time to develop at least 2 episodes of YZ (p = 0.03). The airway microbiota have changed from randomization to YZ. A switch from the Corynebacterium + Dolosigranulum cluster at randomization to the Moraxella- cluster at YZ poses the highest risk of severe asthma exacerbation (p = 0.04). Corynebacterium's relative abundance at YZ is inversely associated with severe exacerbation (p = 0.002).


Exon 5 of the p53 gene is a target for deletions in ovarian cancer.
Missense point mutations, leading to inactivation of the p53 tumor suppressor gene product, are currently the most frequent alterations in human cancer. Little, however, is known about small intragenic deletions or insertions occurring in this locus of chromosome 17. We have analyzed 56 primary ovarian tumors for the presence of such abnormalities. The analysis was based on multiplex PCR amplification of exons 1 through 11 of the p53 gene and fragment analysis of the generated PCR products. Mutations were detected in 14% (8 of 56) of the tumors. Deletions were much more prevalent than insertions (seven vs one). Six of the deletions and the insertion affected exon 5, and the other deletion was in exon 7. Two deletions and the insertion did not disrupt the reading frame; the protein product was expressed in the tumor at high concentrations in all three cases. The other five deletions generated a frameshift, which is predicted to result in the production of a truncated protein product. In the case of the deletions, a 2-5-bp repeat was present close to the detected deletion, whereas the insertion duplicated the sequence immediately upstream of the insertion site. Overall our findings indicate that small intragenic p53 deletions/insertions are not rare events in ovarian cancer, and that p53 exon 5 is the target in the vast majority (88%) of the cases.


Genetic screens to identify pathogenic gene variants in the common cancer predisposition Lynch syndrome.
In many individuals suspected of the common cancer predisposition Lynch syndrome, variants of unclear significance (VUS), rather than an obviously pathogenic mutations, are identified in one of the DNA mismatch repair (MMR) genes. The uncertainty of whether such VUS inactivate MMR, and therefore are pathogenic, precludes targeted healthcare for both carriers and their relatives. To facilitate the identification of pathogenic VUS, we have developed an in cellulo genetic screen-based procedure for the large-scale mutagenization, identification, and cataloging of residues of MMR genes critical for MMR gene function. When a residue identified as mutated in an individual suspected of Lynch syndrome is listed as critical in such a reverse diagnosis catalog, there is a high probability that the corresponding human VUS is pathogenic. To investigate the applicability of this approach, we have generated and validated a prototypic reverse diagnosis catalog for the MMR gene MutS Homolog 2 (Msh2) by mutagenizing, identifying, and cataloging 26 deleterious mutations in 23 amino acids. Extensive in vivo and in vitro analysis of mutants listed in the catalog revealed both recessive and dominant-negative phenotypes. Nearly half of these critical residues match with VUS previously identified in individuals suspected of Lynch syndrome. This aids in the assignment of pathogenicity to these human VUS and validates the approach described here as a diagnostic tool. In a wider perspective, this work provides a model for the translation of personalized genomics into targeted healthcare.


Neutrophilic differentiation induced by human upper airway fibroblast-derived granulocyte/macrophage colony-stimulating factor (GM-CSF).
We have established primary lines of fibroblasts from nasal polyp (NP) tissues as well as from normal nasal (NN) mucosa and have examined the ability of these cells to release hormone-like peptide messenger molecules (cytokines). Our results show that human upper airway fibroblasts release granulocyte/macrophage colony-stimulating factor (GM-CSF), granulocyte-CSF (G-CSF), and macrophage-CSF (M-CSF) in vitro. We also show that fibroblasts derived from NP tissue express the gene for GM-CSF at a higher level, and release the GM-CSF product in greater amounts, than NN fibroblasts. In addition, we have examined the ability of these fibroblasts and their conditioned medium (CM) to induce differentiation of human hemopoietic progenitor cells. After 7 d, cultures of these cells in RPMI-10% fetal bovine serum contained 5 +/- 2.5% (mean +/- SD) neutrophils. In contrast, culture of progenitor cells with fibroblasts resulted in significantly greater neutrophilic differentiation (18 +/- 4%). Culture in fibroblast-CM induced a similar degree of differentiation, and fibroblast-CM from NP fibroblasts elicited greater differentiation compared to CM from NN fibroblasts (17.5 +/- 3 versus 12 +/- 3%). The neutrophilic differentiation induced by fibroblast-CM can be fully inhibited by preincubating this CM with a monoclonal neutralizing antibody to human GM-CSF. Thus, our results demonstrate: (1) the ability of human upper airway fibroblasts to release GM-, G-, and M-CSF in vitro; (2) that fibroblasts derived from NP tissues express the gene and release the product GM-CSF at greater levels compared to NN fibroblasts; and (3) that fibroblast-derived GM-CSF causes neutrophilic differentiation of human hemopoietic progenitors.


Allelic diversity for neutral markers retains a higher adaptive potential for quantitative traits than expected heterozygosity.
The adaptive potential of a population depends on the amount of additive genetic variance for quantitative traits of evolutionary importance. This variance is a direct function of the expected frequency of heterozygotes for the loci which affect the trait (QTL). It has been argued, but not demonstrated experimentally, that long-term response to selection is more dependent on QTL allelic diversity than on QTL heterozygosity. Conservation programmes, aimed at preserving this variation, usually rely on neutral markers rather than on quantitative traits for making decisions on management. Here, we address, both through simulation analyses and experimental studies with Drosophila melanogaster, the question of whether allelic diversity for neutral markers is a better indicator of a high adaptive potential than expected heterozygosity. In both experimental and simulation studies, we established synthetic populations for which either heterozygosity or allelic diversity was maximized using information from QTL (simulations) or unlinked neutral markers (simulations and experiment). The synthetic populations were selected for the quantitative trait to evaluate the evolutionary potential provided by the two optimization methods. Our results show that maximizing the number of alleles of a low number of markers implies higher responses to selection than maximizing their heterozygosity.


Esterase 6 and reproduction in Drosophila melanogaster.
A nonspecific carboxylesterase (esterase 6) of Drosophila melanogaster shows greater activity in adult males than in females and is highly concentrated in the anterior ejaculatory duct of the reproductive tract of the male. Esterase 6 is depleted in males by copulation and is transferred to females early during copulation as a component of the seminal fluid. That esterase 6 may be involved in a system controlling the timing of remating is suggested by differences in the activity of this enzyme in a strain of Drosophila selected for a decrease in time to remating and by differences in the timing of remating in females initially inseminated by males lacking or having active esterase 6.