Evidence-based guideline summary: evaluation, diagnosis, and management of congenital muscular dystrophy: Report of the Guideline Development Subcommittee of the American Academy of Neurology and the Practice Issues Review Panel of the American Association of Neuromuscular & Electrodiagnostic Medicine.
To delineate optimal diagnostic and therapeutic approaches to congenital muscular dystrophy (CMD) through a systematic review and analysis of the currently available literature. Relevant, peer-reviewed research articles were identified using a literature search of the MEDLINE, EMBASE, and Scopus databases. Diagnostic and therapeutic data from these articles were extracted and analyzed in accordance with the American Academy of Neurology classification of evidence schemes for diagnostic, prognostic, and therapeutic studies. Recommendations were linked to the strength of the evidence, other related literature, and general principles of care. The geographic and ethnic backgrounds, clinical features, brain imaging studies, muscle imaging studies, and muscle biopsies of children with suspected CMD help predict subtype-specific diagnoses. Genetic testing can confirm some subtype-specific diagnoses, but not all causative genes for CMD have been described. Seizures and respiratory complications occur in specific subtypes. There is insufficient evidence to determine the efficacy of various treatment interventions to optimize respiratory, orthopedic, and nutritional outcomes, and more data are needed regarding complications. Multidisciplinary care by experienced teams is important for diagnosing and promoting the health of children with CMD. Accurate assessment of clinical presentations and genetic data will help in identifying the correct subtype-specific diagnosis in many cases. Multiorgan system complications occur frequently; surveillance and prompt interventions are likely to be beneficial for affected children. More research is needed to fill gaps in knowledge regarding this category of muscular dystrophies.


[The Unified National Health System and public policies: psychological care for menopausal women in the Federal District, Brazil].
This study focused on psychological care for middle-aged women in public health services in the Federal District (Brasilia), Brazil. The article discusses women's health and more specifically menopause and its place in Brazilian public health policies. The survey confirmed the lack of psychological support for menopausal women. In most cases only outpatient medical care was offered. No psychologist had been designated in any of the units surveyed to assist these women. The study concludes that this period of women's life has failed to receive psychological care in Brazil, thus neglecting the principles of the Unified National Health System. Menopausal women deserve comprehensive health care, including attention to conflicts related to biological, psychological, and socio-cultural dimensions of aging, thus contributing to the process of working through maturity.


Symptom presentation in patients hospitalized with acute heart failure.
The objectives of this study were to examine the type and frequency of symptoms in patients hospitalized with acute heart failure (HF) as well as the relationship between symptom patterns and patient characteristics, treatment practices, and hospital outcomes in patients hospitalized with decompensated HF. The study sample consisted of 4537 residents of the Worcester, MA metropolitan area hospitalized for decompensated HF at 11 greater Worcester medical centers in 1995 and 2000. The average age of the study sample was 76 years; the majority (57%) were women, and three-quarters of our patient population had been previously diagnosed with HF. Dyspnea (93%) was the most frequent complaint reported by patients followed by the presence of peripheral edema (70%), cough (51%), orthopnea (37%), and chest pain/discomfort (30%). Patients reporting few cardiac symptoms were less likely to be treated with effective cardiac therapies during hospitalization than patients with multiple cardiac signs and symptoms and experienced higher hospital (9.7% vs. 7.7%) as well as 30-day (17.1% vs. 10.2%) death rates (P < 0.05). The results of this study in residents of a large New England community suggest that patients with fewer reported symptoms of decompensated HF were less likely to receive effective cardiac treatments and had worse short-term outcomes. Reasons for these differences in treatment practices and short-term outcomes need to be elucidated and attention directed to these high-risk patients.


Bio-inspired polarized skylight-based navigation sensors: a review.
Animal senses cover a broad range of signal types and signal bandwidths and have inspired various sensors and bioinstrumentation devices for biological and medical applications. Insects, such as desert ants and honeybees, for example, utilize polarized skylight pattern-based information in their navigation activities. They reliably return to their nests and hives from places many kilometers away. The insect navigation system involves the dorsal rim area in their compound eyes and the corresponding polarization sensitive neurons in the brain. The dorsal rim area is equipped with photoreceptors, which have orthogonally arranged small hair-like structures termed microvilli. These are the specialized sensors for the detection of polarized skylight patterns (e-vector orientation). Various research groups have been working on the development of novel navigation systems inspired by polarized skylight-based navigation in animals. Their major contributions are critically reviewed. One focus of current research activities is on imitating the integration path mechanism in desert ants. The potential for simple, high performance miniaturized bioinstrumentation that can assist people in navigation will be explored.


Mobile phone radiation causes brain tumors and should be classified as a probable human carcinogen (2A) (review).
Quickly changing technologies and intensive uses of radiofrequency electromagnetic field (RF-EMF)‑emitting phones pose a challenge to public health. Mobile phone users and uses and exposures to other wireless transmitting devices (WTDs) have increased in the past few years. We consider that CERENAT, a French national study, provides an important addition to the literature evaluating the use of mobile phones and risk of brain tumors. The CERENAT finding of increased risk of glioma is consistent with studies that evaluated use of mobile phones for a decade or longer and corroborate those that have shown a risk of meningioma from mobile phone use. In CERENAT, exposure to RF‑EMF from digitally enhanced cordless telephones (DECTs), used by over half the population of France during the period of this study, was not evaluated. If exposures to DECT phones could have been taken into account, the risks of glioma from mobile phone use in CERENAT are likely to be higher than published. We conclude that radiofrequency fields should be classified as a Group 2A ̔probable̓ human carcinogen under the criteria used by the International Agency for Research on Cancer (Lyon, France). Additional data should be gathered on exposures to mobile and cordless phones, other WTDs, mobile phone base stations and Wi‑Fi routers to evaluate their impact on public health. We advise that the as low as reasonable achievable (ALARA) principle be adopted for uses of this technology, while a major cross‑disciplinary effort is generated to train researchers in bioelectromagnetics and provide monitoring of potential health impacts of RF‑EMF.


Human cytomegalovirus-inhibitory flavonoids: studies on antiviral activity and mechanism of action.
We report antiviral activity against human cytomegalovirus for certain dietary flavonoids and their likely biochemical mechanisms of action. Nine out of ten evaluated flavonoids blocked HCMV replication at concentrations that were significantly lower than those producing cytotoxicity against growing or stationary phase host cells. Baicalein was the most potent inhibitor in this series (IC(50)=0.4-1.2 microM), including positive control ganciclovir. Baicalein and genistein were chosen as model compounds to study the antiviral mechanism(s) of action for this series. Both flavonoids significantly reduced the levels of HCMV early and late proteins, as well as viral DNA synthesis. Baicalein reduced the levels of HCMV immediate-early proteins to nearly background levels while genistein did not. The antiviral effects of genistein, but not baicalein, were fully reversible in cell culture. Pre-incubation of concentrated virus stocks with either flavonoid did not inhibit HCMV replication, suggesting that baicalein did not directly inactivate virus particles. Baicalein functionally blocked epidermal growth factor receptor tyrosine kinase activity and HCMV nuclear translocation, while genistein did not. At 24h post infection HCMV-infected cells treated with genistein continued to express immediate-early proteins and efficiently phosphorylate IE1-72. However, HCMV induction of NF-kappaB and increases in the levels of cell cycle regulatory proteins--events that are associated with immediate-early protein functioning--were absent. The data suggested that the primary mechanism of action for baicalein may be to block HCMV infection at entry while the primary mechanism of action for genistein may be to block HCMV immediate-early protein functioning.


Inverse PPARβ/δ agonists suppress oncogenic signaling to the ANGPTL4 gene and inhibit cancer cell invasion.
Besides its established functions in intermediary metabolism and developmental processes, the nuclear receptor peroxisome proliferator-activated receptor β/δ (PPARβ/δ) has a less defined role in tumorigenesis. In the present study, we have identified a function for PPARβ/δ in cancer cell invasion. We show that two structurally divergent inhibitory ligands for PPARβ/δ, the inverse agonists ST247 and DG172, strongly inhibit the serum- and transforming growth factor β (TGFβ)-induced invasion of MDA-MB-231 human breast cancer cells into a three-dimensional matrigel matrix. To elucidate the molecular basis of this finding, we performed chromatin immunoprecipitation sequencing (ChIP-Seq) and microarray analyses, which identified the gene encoding angiopoietin-like 4 (ANGPTL4) as the major transcriptional PPARβ/δ target in MDA-MB-231 cells, previously implicated in TGFβ-mediated tumor progression and metastatic dissemination. We show that the induction of ANGPTL4 by TGFβ and other oncogenic signals is strongly repressed by ST247 and DG172 in a PPARβ/δ-dependent fashion, resulting in the inhibition of ANGPTL4 secretion. This effect is attributable to these ligands' ability to induce a dominant transcriptional repressor complex at the site of transcription initiation that blocks preinitiation complex formation through an histone deacetylase-independent, non-canonical mechanism. Repression of ANGPTL4 transcription by inverse PPARβ/δ agonists is functionally linked to the inhibition of cancer cell invasion into a three-dimensional matrix, as (i) invasion of MDA-MB-231 cells is critically dependent on ANGPTL4 expression, (ii) recombinant ANGPTL4 stimulates invasion, and (iii) reverses the inhibitory effect of ST247 and DG172. These findings indicate that a PPARβ/δ-ANGPTL4 pathway is involved in the regulation of tumor cell invasion and that its pharmacological manipulation by inverse PPARβ/δ agonists is feasible.


Spawning segregation and philopatry are major prezygotic barriers in sympatric cryptic Mugil cephalus species.
The flathead mullet, Mugil cephalus, is a commercially vital fish in fisheries and aquaculture worldwide. Genetic analyses have recently revealed three cryptic species of M. cephalus in the Northwest Pacific. These species are sympatric in Taiwanese waters and specific reproductive behaviors have been suggested to be a major prezygotic barrier. Species composition was evaluated in samples of M. cephalus at different growth stages collected from various habitats (offshore spawning ground, estuarine nursery and feeding areas) over several months or years. The gonadosomatic index of adults and the body length of juveniles were recorded to determine the reproductive season and recruitment periods in estuaries. The results revealed partially temporal spawning isolation between species pairs, spatial segregation on specific spawning grounds and strong philopatry preclude hybridization. Thus, the results imply that traditional fisheries of mature fish in the Taiwan Strait target only one species, whereas aquaculture in Taiwan contain juveniles of all three species collected in estuaries. The ecological niche and demography of these species must be investigated further to estimate the impact of juvenile sources on aquaculture.


Semidefinite programming for model-based sensorless adaptive optics.
Wavefront sensorless adaptive optics methodologies are widely considered in scanning fluorescence microscopy where direct wavefront sensing is challenging. In these methodologies, aberration correction is performed by sequentially changing the settings of the adaptive element until a predetermined image quality metric is optimized. An efficient aberration correction can be achieved by modeling the image quality metric with a quadratic polynomial. We propose a new method to compute the parameters of the polynomial from experimental data. This method guarantees that the quadratic form in the polynomial is semidefinite, resulting in a more robust computation of the parameters with respect to existing methods. In addition, we propose an algorithm to perform aberration correction requiring a minimum of N+1 measurements, where N is the number of considered aberration modes. This algorithm is based on a closed-form expression for the exact optimization of the quadratic polynomial. Our arguments are corroborated by experimental validation in a laboratory environment.


Development of tumor-infiltrating CD8+ T cell memory precursor effector cells and antimelanoma memory responses are the result of vaccination and TGF-β blockade during the perioperative period of tumor resection.
A main goal of cancer immunology research is the formation of Ag-specific memory T cell immunity capable of activation upon tumor re-encounter. The requirements necessary to overcome the inhibitory signals present in the tumor microenvironment and form such memory T cell responses are unknown. In contrast to previous studies targeting tumors expressing highly immunogenic model Ags, we demonstrate that alleviating tumor-induced suppression along with vaccination against authentic Ags during the perioperative period provides long-lasting protection against a highly suppressive and poorly immunogenic melanoma. In this study, we employed DNA vaccination with an immunologically optimized mouse melanoma-shared Ag, Trp1ee/ng, combined with systemic TGF-β blockade during the perioperative period of primary tumor resection, to confer protection against B16 melanoma, and against JBRH, an independently derived melanoma unrelated to B16. Importantly, we demonstrate that correlative to memory responses, perioperative immunotherapy increases the formation of tumor-infiltrating and tumor-reactive CD8(+) T cells expressing low levels of the transcription factor T-bet, defined as memory precursor effector cells. We show that conditions for an immunologically fertile environment are met when TGF-β blockade and vaccination are applied during the perioperative period of primary tumor resection. These findings address limitations of current CD8(+) T cell immunotherapies against cancer by generating effective CD8(+) T cell memory recall responses.


Does high and intensive care reduce coercion? Association of HIC model fidelity to seclusion use in the Netherlands.
A new inpatient care model has been developed in the Netherlands: High and Intensive Care (HIC). The purpose of HIC is to improve quality of inpatient mental healthcare and to reduce coercion. In 2014, audits were held at 32 closed acute admission wards for adult patients throughout the Netherlands. The audits were done by trained auditors, who were professionals of the participating institutes, using the HIC monitor, a model fidelity scale to assess implementation of the HIC model. The HIC model fidelity scale (67 items) encompasses 11 domains including for example team structure, team processes, diagnostics and treatment, and building environment. Data on seclusion and forced medication was collected using the Argus rating scale. The association between HIC monitor scores and the use of seclusion and forced medication was analyzed, corrected for patient characteristics. Results showed that wards having a relatively high HIC monitor total score, indicating a high level of implementation of the model as compared to wards scoring lower on the monitor, had lower seclusion hours per admission hours (2.58 versus 4.20) and less forced medication events per admission days (0.0162 versus 0.0207). The HIC model fidelity scores explained 27% of the variance in seclusion rates (p < 0.001). Adding patient characteristics to HIC items in the regression model showed an increase of the explained variance to 40%. This study showed that higher HIC model fidelity was associated with less seclusion and less forced medication at acute closed psychiatric wards in the Netherlands.


[Expression profile of clock genes during the process of embryonic stem cells differentiation].
To analysis the expression profile of7 clock genes during mouse embryonic stem cell (mES) differentiation. Mouse ES cells of the line 129 were cultured and induced to differentiate into neurons by 5 stages method. The expression of 7 clock genes: BMAL1, CLOCK, CRY1, CRY2, PER1, PER2, and PER3 in the five stages were determined by RT-PCR. In the amplification condition, no non-specific band was found, and only bands of expected sizes were detected. Six clock genes, BMAL1, CLOCK, CRY1, CRY2, PER1 and PER3 were expressed in 129 cells at all five stages in the differentiation process, however, PER2, could only be determined in the last stage. The transcription profile of the clock genes is different during the differentiation of the ES cells towards neurons. The transcription of the gene PER2 is limited to the final stage, when postmitotic neurons and astrocytes emerged. A clock genes loop existing in the mature cells has not been established during the process of ES cell differentiation.


Interleukin 10 suppresses tumor growth and metastasis of human melanoma cells: potential inhibition of angiogenesis.
Interleukin 10 (IL-10) inhibits the production of a wide range of cytokines in various cell types. The purpose of this study was to determine whether the expression of the IL-10 gene can influence tumor growth and metastatic properties of human melanoma cells. The human melanoma cell line, A375P, which does not produce endogenous IL-10, was transfected with a hygromycin expression vector (control) or a vector containing full-length murine IL-10 cDNA. A375P parental cells, A375P-Hygro, and A375P-IL-10-positive cells were injected s.c. and i.v. into nude mice. A375P-IL-10 cells produced significantly slower growing s.c. tumors and fewer lung metastases than control cells. The tumorigenicity of the human melanoma A375SM and the murine melanoma B16-BL6 cells was also significantly inhibited when they were admixed with A375P-IL-10 but not with A375P-Hygro before s. c. injection into nude mice. The suppression of tumor growth and metastasis was directly correlated with a decrease in neovascularity determined by immunostaining with anti-factor VIII. Because tumor-associated macrophages are the major source of angiogenic molecules in melanoma, we used reverse transcription-PCR to demonstrate that IL-10 down-regulates the production of vascular endothelial growth factor, the most potent angiogenic factor in activated macrophages. Other factors involved in angiogenesis such as IL-1beta, tumor necrosis factor-alpha, IL-6, and the proteinase matrix metalloproteinase-9 were also inhibited in activated macrophages by supernatants from A375P-IL-10 cells. Collectively, these data suggest that the production of IL-10 by tumor cells inhibits macrophages-derived angiogenic factors, and hence, tumor growth and metastasis.


[Effect of fertilization combinations of nitrogen, phosphorus, and potassium on four phenolic compounds of cultivated Rhodiola crenulata].
This study is aimed to explore the effect of nitrogen, phosphorus and potassium combined application on the active components of Rhodiola crenulata. R. crenulata was used as the research object, "3414" fertilization experiment were conducted with regular fertilization of NPK(N 60 kg·hm⁻², P₂O₅ 100 kg·hm⁻²,KCl 160 kg·hm⁻²) to study the effect of different rates of NPK fertilization on the total amount of 4 phenolic constituents of gallic acid, salidroside, tyrol and ethyl gallate through field test. The results show that the content of salidroside was higher in the treatment of N₁P₂K₁ and N₁P₂K₂, andthe total amount of four phenols was higher in the treatment of N₁P₂K₂ and N₂P₂K₂. The suitable level of nitrogen, phosphorus and potassium promoted the accumulation of the 4 kinds of phenols.The amount of fertilizer recommended by the three factor fertilizer effect equation,(N 0 kg·hm⁻²,P₂O₅ 150 kg·hm⁻²,KCl 31.71 kg·hm⁻²) obtained the highest content of salidroside, and it was 1.54%.(N 35.54 kg·hm⁻²,P₂O₅ 150 kg·hm⁻²,KCl 237.73 kg·hm⁻²)obtained the highest content of 4 kinds of phenolic compounds, and it was 1.93%. This study provides a reference for the standardization of artificial planting of endangered Tibetan medicine.


Oxygen saturation measurements in canine blood containing hemoglobin glutamer-200 (Bovine): in vitro validation of the NOVA CO-Oximeter.
This study was designed to validate in vitro oxygen saturation (SO2) measurements with the NOVA CO-Oximeter (Nova Biomedical Corp, Waltham, Mass, USA) in canine blood containing hemoglobin (Hb) glutamer-200 bovine (Hb-200; Oxyglobin, Biopure, Cambridge, Mass, USA) as a Hb-based oxygen carrier recently introduced into clinical practice. In the first set of experiments, stored blood from 6 mixed-breed canine blood donors was used. Target PO2 levels were reached in aliquots of blood samples by tonometry. Oxygen saturation was then measured with the test device and calculated based on known PO2 values. In the second set of experiments, total oxygen content was directly measured by means of an oxygen-specific electrode in aliquots of fresh whole arterial, venous, and mixed (arterial-venous) blood withdrawn from the same canine blood donors. Hb-200 was added to those blood samples to yield plasma Hb concentrations of 1.62, 3.25, 6.50, and 9.75 g/dL. Based on Hb content and SO2 measured by the NOVA CO-Oximeter in these samples, total oxygen content was also calculated for each sample and compared with measured values. A strong correlation was found between SO2 values measured with the co-oximeter in samples after tonometry, and calculated SO2 based on known PO2. Directly measured total blood O2 content varied by </=5% from values computed based on co-oximeter measurements of Hb content and SO2. These results did not change with different levels of oxygenation of the samples or different plasma Hb-200 concentrations. In conclusion, the NOVA CO-Oximeter is an accurate analyzer for measurement of SO2 after Hb-200 administration to canine blood.


Parental child-rearing attitudes are associated with functional constipation in childhood.
Parenting factors are assumed to play a role in the development and maintenance of childhood constipation. However, knowledge about the association between parenting factors and childhood constipation is limited. This study investigates the association between parental child-rearing attitudes and prominent symptoms of functional constipation and assesses the strength of this association. Cross-sectional data of 133 constipated children and their parents were collected. The gastrointestinal outpatient clinic at the Emma Children's Hospital in the Netherlands. Children with functional constipation aged 4-18 years referred by general practitioners, school doctors and paediatricians. Parental child-rearing attitudes were assessed by the Amsterdam version of the Parental Attitude Research Instrument (A-PARI). Symptoms of constipation in the child were assessed by a standardised interview. Negative binomial and logistic regression models were used to test the association between child-rearing attitudes and constipation symptoms. Parental child-rearing attitudes are associated with defecation and faecal incontinence frequency. Higher and lower scores on the autonomy attitude scale were associated with decreased defecation frequency and increased faecal incontinence. High scores on the overprotection and self-pity attitude scales were associated with increased faecal incontinence. More and stronger associations were found for children aged ≥6 years than for younger children. Parental child-rearing attitudes are associated with functional constipation in children. Any parenting issues should be addressed during treatment of children with constipation. Referral to mental health services is needed when parenting difficulties hinder treatment or when the parent-child relationship is at risk. ISRCTN2518556.


Secretory Vesicle Polar Sorting, Endosome Recycling and Cytoskeleton Organization Require the AP-1 Complex in Aspergillus nidulans.
The AP-1 complex is essential for membrane protein traffic via its role in the pinching-off and sorting of secretory vesicles (SVs) from the trans-Golgi and/or endosomes. While its essentiality is undisputed in metazoa, its role in simpler eukaryotes seems less clear. Here, we dissect the role of AP-1 in the filamentous fungus Aspergillus nidulans and show that it is absolutely essential for growth due to its role in clathrin-dependent maintenance of polar traffic of specific membrane cargoes toward the apex of growing hyphae. We provide evidence that AP-1 is involved in both anterograde sorting of RabERab11-labeled SVs and RabA/BRab5-dependent endosome recycling. Additionally, AP-1 is shown to be critical for microtubule and septin organization, further rationalizing its essentiality in cells that face the challenge of cytoskeleton-dependent polarized cargo traffic. This work also opens a novel issue on how nonpolar cargoes, such as transporters, are sorted to the eukaryotic plasma membrane.


Identification of trichotoxin, a novel chlorinated compound associated with the bloom forming Cyanobacterium, Trichodesmium thiebautii.
Trichodesmium is a suspected toxin-producing nonheterocystous cyanobacteria ubiquitous in tropical, subtropical, and temperate seas. The genus is known for its ability to fix nitrogen and form massive blooms. In oligotrophic seas, it can dominate the biomass and be a major component of oceanic primary production and global nitrogen cycling. Numerous reports suggest Trichodesmium-derived toxins are a cause of death of fish, crabs, and bivalves. Laboratory studies have demonstrated neurotoxic effects in T. thiebautii cell extracts and field reports suggest respiratory distress and contact dermatitis of humans at collection sites. However, Trichodesmium toxins have not been identified and characterized. Here, we report the extraction of a lipophilic toxin from field-collected T. thiebautii using a purification method of several chromatographic techniques, nuclear magnetic resonance (NMR), mass spectroscopy (MS), and Fourier transformed-infrared spectroscopy (FT-IR). Trichotoxin has a molecular formula of C(20)H(27)ClO and a mass of 318 m/z and possesses cytotoxic activity against GH(4)C(1) rat pituitary and Neuro-2a mouse neuroblastoma cells. A detection method using liquid chromatography/mass spectrometry (LC/MS) was developed. This compound is the first reported cytotoxic natural product isolated and fully characterized from a Trichodesmium species.


RNA-templated DNA repair.
RNA can act as a template for DNA synthesis in the reverse transcription of retroviruses and retrotransposons and in the elongation of telomeres. Despite its abundance in the nucleus, there has been no evidence for a direct role of RNA as a template in the repair of any chromosomal DNA lesions, including DNA double-strand breaks (DSBs), which are repaired in most organisms by homologous recombination or by non-homologous end joining. An indirect role for RNA in DNA repair, following reverse transcription and formation of a complementary DNA, has been observed in the non-homologous joining of DSB ends. In the yeast Saccharomyces cerevisiae, in which homologous recombination is efficient, RNA was shown to mediate recombination, but only indirectly through a cDNA intermediate generated by the reverse transcriptase function of Ty retrotransposons in Ty particles in the cytoplasm. Although pairing between duplex DNA and single-strand (ss)RNA can occur in vitro and in vivo, direct homologous exchange of genetic information between RNA and DNA molecules has not been observed. We show here that RNA can serve as a template for DNA synthesis during repair of a chromosomal DSB in yeast. The repair was accomplished with RNA oligonucleotides complementary to the broken ends. This and the observation that even yeast replicative DNA polymerases such as alpha and delta can copy short RNA template tracts in vitro demonstrate that RNA can transfer genetic information in vivo through direct homologous interaction with chromosomal DNA.


Recruitment of knee joint ligaments.
On the basis of earlier reported data on the in vitro kinematics of passive knee-joint motions of four knee specimens, the length changes of ligament fiber bundles were determined by using the points of insertion on the tibia and femur. The kinematic data and the insertions of the ligaments were obtained by using Roentgenstereophotogrammetry. Different fiber bundles of the anterior and posterior cruciate ligaments and the medial and lateral collateral ligaments were identified. On the basis of an assumption for the maximal strain of each ligament fiber bundle during the experiments, the minimal recruitment length and the probability of recruitment were defined and determined. The motions covered the range from extension to 95 degrees flexion and the loading conditions included internal or external moments of 3 Nm and anterior or posterior forces of 30 N. The ligament length and recruitment patterns were found to be consistent for some ligament bundles and less consistent for other ligament bundles. The most posterior bundle of each ligament was recruited in extension and the lower flexion angles, whereas the anterior bundle was recruited for the higher flexion angles. External rotation generally recruited the collateral ligaments, while internal rotation recruited the cruciate ligaments. However, the anterior bundle of the posterior cruciate ligament was recruited with external rotation at the higher flexion angles. At the lower flexion angles, the anterior cruciate and the lateral collateral ligaments were recruited with an anterior force. The recruitment of the posterior cruciate ligament with a posterior force showed that neither its most anterior nor its most posterior bundle was recruited at the lower flexion angles. Hence, the posterior restraint must have been provided by the intermediate fiber bundles, which were not considered in the experiment. At the higher flexion angles, the anterior bundles of the anterior cruciate ligament and the posterior cruciate ligament were found to be recruited with anterior and posterior forces, respectively. The minimal recruitment length and the recruitment probability of ligament fiber bundles are useful parameters for the evaluation of ligament length changes in those experiments where no other method can be used to determine the zero strain lengths, ligament strains and tensions.


Detection of Aspergillus flavus in soil by immunofluorescent staining.
Strains of Aspergillus flavus grown in soil in the presence of buried slides could be detected on the slides by fluorescent antibody techniques. Staining with A. flavus antiserum labeled with fluorescein, followed by examination with fluorescence microscopy revealed characteristic fluorescence at sites of distribution of the homologous fungus. The specificity of the reaction and the absence of nonspecific absorption of antibody to soil materials suggest that the method may be useful in studying the ecology of the soil microflora.


[Clinical staging and early intervention for borderline personality disorder].
Within the accumulating evidence for a life span perspective on borderline personality disorder (BPD), the key factor is a developmental perspective.
 AIM: To demonstrate that the lessons learnt from early intervention in somatic medicine and psychosis should be used to improve the diagnosis and treatment of BPD.
 METHOD: We describe the rationale for early detection and intervention and present a staging model which can serve as a guideline for the development and selection of interventions for BPD.
 RESULTS: There is increasing evidence that BPD first manifests itself in adolescence and that BPD symptoms can already be distinguished from normative adolescent development. BPD tends to develop gradually and to have a progressive, social and professional impact. Inadequate treatment can lead to iatrogenic damage, whereas adequate treatment shows promising results in adolescents with emerging BPD. These findings may underpin an early intervention paradigm for BPD.
 CONCLUSION: Early intervention and clinical staging can improve the assessment and treatment of severe forms of psychopathology, such as BPD. This has implications for research, mental health care policy and society.


Adaptive optics via self-interference digital holography for non-scanning three-dimensional imaging in biological samples.
Three-dimensional imaging in biological samples usually suffers from performance degradation caused by optical inhomogeneities. Here we proposed an approach to adaptive optics in fluorescence microscopy where the aberrations are measured by self-interference holographic recording and then corrected by a post-processing optimization procedure. In our approach, only one complex-value hologram is sufficient to measure and then correct the aberrations, which results in fast acquisition speed, lower exposure time, and the ability to image in three-dimensions without the need to scan the sample or any other element in the system. We show proof-of-principle experiments on a tissue phantom containing fluorescence particles. Furthermore, we present three-dimensional reconstructions of actin-labeled MCF7 breast cancer cells, showing improved resolution after the correction of aberrations. Both experiments demonstrate the validity of our method and show the great potential of non-scanning adaptive three-dimensional microscopy in imaging biological samples with improved resolution and signal-to-noise ratio.


Impact of Different Estimation Methods on Obesity-Attributable Mortality Levels and Trends: The Case of The Netherlands.
The available methodologies to estimate the obesity-attributable mortality fraction (OAMF) affect the levels found and hamper the construction of time series. Our aim was to assess the impact of using different techniques to estimate the levels and the trends in obesity-attributable mortality for The Netherlands between 1981 to 2013. Using Body Mass Index (BMI), all-cause and cause-specific mortality data, and worldwide and European relative risks (RRs), we estimated OAMFs using three all-cause approaches (partially adjusted, weighted sum, and the two combined) and one cause-of-death approach (Comparative Risk Assessment; CRA). We adjusted the CRA approach to purely capture obesity (BMI ≥ 30 kg/m²). The different approaches led to a range of estimates. The weighted sum method using worldwide RRs generated the lowest (0.9%) while the adjusted CRA approach using 2013 RRs generated the highest estimate (1.5%). Using European-specific RRs instead of worldwide RRs resulted in higher estimates. Most of the approaches revealed an increasing OAMF over the period 1981 to 2013 especially from 1993 onwards except for the adjusted CRA approach among women. Estimates of OAMF levels and trends differed depending on the method applied. Given the limited available data, we recommend using the weighted-sum method to compare obesity-attributable mortality across European countries over time.


Regulation of lateral flagella gene transcription in Vibrio parahaemolyticus.
Two distinctly different organelles of locomotion are produced by Vibrio parahaemolyticus. The polar flagellum is responsible for motility in a liquid environment (swimming), and the lateral flagella enable the bacteria to move over surfaces (swarming). Synthesis of lateral flagella occurs when V. parahaemolyticus is grown on agar media but not when it is grown in liquid media. We used lux (luminescence gene) fusions to conveniently and sensitively analyze the factors which influence transcription of lateral flagella genes (laf). Transposon mini-Mu lux was used to mutagenize V. parahaemolyticus and to generate laf::lux transcriptional fusions. Mutants with insertions of mini-Mu lux in laf genes were defective in the swarming phenotype and produced light when the bacteria were propagated on agar media, but not when cells were grown in liquid media. Thus, surface-dependent expression of lateral flagella synthesis is controlled by regulation of transcription. Such fusion strains were also used to further define the environmental conditions which induce laf gene expression. Cultivation on media solidified by gelling agents other than agar also induced light production in fusion strains, as did growth on a variety of hydrophilic membrane filters suspended over liquid media. Growth at an air-surface interface was not necessary for expression since embedding the fusion strains in agar was also effective. Furthermore, induction of laf gene transcription could also be accomplished by increasing the viscosity of the liquid medium by the addition of a high-molecular-weight polymer such as polyvinylpyrrolidone. Increase in luminescence of the fusion strains was detected within 30 min of initiation of the inducing circumstance, and reversal of induction, e.g., by dilution of the viscous medium, resulted in a rapid decline in the rate of increase in luminescence. Conditions that induced luminescence in the fusion strains also induced the synthesis of lateral flagella in wild-type V. parahaemolyticus. The growth environment of the genes, and it appears that the signal that triggers laf expression is physical rather than chemical in nature. Possibilities for a sensing mechanism are discussed.


Influence of serum and albumin on the in vitro anandamide cytotoxicity toward C6 glioma cells assessed by the MTT cell viability assay: implications for the methodology of the MTT tests.
Anandamide (AEA), an endogenous ligand of cannabinoid CB1 and CB2 receptors, which also binds transient receptor potential vanilloid type 1 receptor (TRPV1), has been shown to display substantial selective cytotoxicity toward some cancer cell lines in vitro, although the relevant data are not consistent. In the present study, we employed the MTT test to assess short-term cytotoxicity of AEA on C6 rat glioma cell culture. When anandamide was administered to the culture medium with foetal bovine serum (FBS), no cytotoxic effect was observed following 24 h exposure of the glioma cells to micromolar concentrations of AEA. However, if no serum was present in the medium, micro-to-submicromolar concentrations of AEA induced dose-dependent cytotoxicity clearly detectable after 24 h. Control experiments made it possible to exclude significant interference of serum with the MTT test per se. Bovine serum albumin mimicked the effect of FBS. We conclude that the apparent inhibition of short-term cytotoxicity of AEA toward C6 rat glioma cells in vitro is caused by binding AEA to serum proteins such as albumin. Taking into account that blood serum or albumin is practically always present in cell culture media, we discuss implications of binding substances to serum proteins for methodology and interpretation of in vitro cytotoxicity testing.


Time trend analysis of return to work after stroke in Denmark 1996-2006.
In the period 1997-2005, the Danish government initiated a series of legislative changes aimed at facilitating RTW (return to work) in the Danish population. In the present study, we investigated the odds of being gainfully occupied ca. two years after stroke as a function of onset calendar year, 1996-2006. All previously employed 20-57 year-old stroke patients in Denmark 1996-2006 (N = 19985) were followed prospectively through national registers. The analysis was controlled for the type of stroke and a series of demographic, structural and occupational variables. The odds for RTW increased significantly during the study period (P < 0.0001). The odds at the end of the period were more than twice as high as they were at the beginning, even after post hoc control for improved survival and decreased unemployment rates. The most conspicuous increase coincided with a change in the sickness benefit act that took place in 2005. The study provides quite strong circumstantial evidence that the legislative changes had an effect on the odds of return to work after stroke. More direct evidence is desirable, but such can only be obtained through a randomized controlled study.


Effects of role typicality on processing person information in German: evidence from an ERP study.
The present ERP study investigated how reference resolution is affected by semantic cues to referent gender such as the gender typicality of role names. Participants read general statements about social and occupational groups denoted by a role name (e.g., pilots, florists), which was followed by a co-referring NP with masculine, feminine or neutral lexical gender (e.g., these men/women/people) that could either semantically match, mismatch or be neutral to the role name's gender typicality. The study was run in German. Between 300 and 400 ms (N400) responses to all anaphors were more negative after typically male than after typically female antecedents. Between 500 and 700 ms (P600) role name typicality interacted with the lexical gender of the anaphor. Responses to feminine anaphors were more positive in case of incongruent compared to congruent typicality of the antecedent. The effects suggest that participants anticipate a possible semantic incongruity before the anaphor is actually resolved. They, furthermore show that different cues to referent gender are conceptually integrated after 500 ms following the onset of the anaphoric noun. Results are discussed with regard to two-stage models of reference resolution.


How does MyPyramid compare to other population-based recommendations for controlling chronic disease?
The Department of Agriculture's MyPyramid is presented as food guidance for the US general public and not "a therapeutic diet for any specific health condition," although many adults in this country are overweight or obese and many experience diet-related disorders. This paper shows the recommendations in MyPyramid are remarkably consistent with the various recommendations to control obesity and diabetes, heart disease and stroke, hypertension, cancer, and osteoporosis. Specifically, the food intake recommendations are similar to those recommended by the Dietary Approaches to Stop Hypertension Eating Plan, the American Heart Association, and the American Cancer Society; plus, the calculated nutrient intakes associated with following the guide are generally within the ranges of nutrient recommendations from the Clinical Guidelines on Overweight and Obesity, the American Diabetes Association, the National Cholesterol Education Program, the American Heart Association, the National Committee on High Blood Pressure, and the American Institute for Cancer Research. However, for actual nutrient levels to conform to dietary guidance, key assumptions regarding how closely individuals will follow the MyPyramid patterns must be made: an appropriate energy level must be selected and adhered to, and an appropriate profile of foods must be selected. These issues must be understood by food and nutrition professionals and disseminated to the public for MyPyramid to reach its potential.


The Association Between Some Macro and Trace Elements in Saliva and Periodontal Status.
Changes in the macro and trace element composition of saliva might be indicative for pathological changes in periodontal tissues. However, there is a lack of evidence in the literature showing associations between mineral elements and periodontal status. The aim of this study was to determine whether such associations occur. Totally, 190 systemically healthy non-smoker participants (mean age 32.2 ± 6.02; 50 periodontally healthy, 50 gingivitis, 50 chronic periodontitis, and 40 aggressive periodontitis individuals) were included in this cross-sectional study. Salivary levels of some macro and trace elements were measured by using inductively coupled plasma mass spectrometry (ICP-MS). Kruskal-Wallis's test was used for statistical analysis. Statistically significant differences were found in sodium (Na), magnesium (Mg), potassium (K), calcium (Ca), vanadium (V), chromium Cr), manganese (Mn), iron (Fe), rubidium (Rb), strontium (Sr), and selenium (Se) concentrations among the groups. Significant increases in the essential minerals Na, Mg, K, Ca, Fe, and Se occurred in both periodontitis groups when compared to the gingivitis and periodontally healthy groups. Lower Se, Sr, Fe, Mn, and V concentrations were found in the aggressive periodontitis group than in the chronic periodontitis group. The results of this study demonstrated that assessment of mineral element concentrations in saliva might be useful in assessing periodontal health and disease. However, further studies are required to determine whether the change in a specific mineral element is the result of periodontal disease or is involved in its pathogenesis.


Hydrodynamic coupling of particle inclusions embedded in curved lipid bilayer membranes.
We develop theory and computational methods to investigate particle inclusions embedded within curved lipid bilayer membranes. We consider the case of spherical lipid vesicles where inclusion particles are coupled through (i) intramembrane hydrodynamics, (ii) traction stresses with the external and trapped solvent fluid, and (iii) intermonolayer slip between the two leaflets of the bilayer. We investigate relative to flat membranes how the membrane curvature and topology augment hydrodynamic responses. We show how both the translational and rotational mobility of protein inclusions are effected by the membrane curvature, ratio of intramembrane viscosity to solvent viscosity, and intermonolayer slip. For general investigations of many-particle dynamics, we also discuss how our approaches can be used to treat the collective diffusion and hydrodynamic coupling within spherical bilayers.


Trefoil factor-1 upregulation in estrogen-receptor positive breast cancer correlates with an increased risk of bone metastasis.
Bone is one of the most preferred sites of metastatic spread from different cancer types, including breast cancer. However, different breast cancer subtypes exhibit distinct metastatic behavior in terms of kinetics and anatomic sites of relapse. Despite advances in the diagnosis, the identification of patients at high-risk of bone recurrence is still an unmet clinical need. We conducted a retrospective analysis, by gene expression and immunohistochemical assays, on 90 surgically resected breast cancer samples collected from patients who experienced no evidence of distant metastasis, bone or visceral metastasis in order to identify a primary tumor-derived marker of bone recurrence. We identified trefoil factor-1 (pS2 or TFF1) as strictly correlated to bone metastasis from ER+ breast cancer. In silico analysis was carried out to confirm this observation, linking gene expression data with clinical characteristics available from public clinical datasets. Then, we investigated TFF1 function in ER+ breast cancer tumorigenesis and bone metastasis through xenograft in vivo models of MCF 7 breast cancer with gain and loss of function of TFF1. As a response to microenvironmental features in primary tumors, TFF1 expression could modulate ER+ breast cancer growth, leading to a less proliferative phenotype. Our results showed it may not play a role in late stages of bone metastasis, however further studies are warranted to understand whether it could contribute in the early-stages of the metastatic cascade. In conclusion, TFF1 upregulation in primary ER+ breast cancer could be useful to identify patients at high-risk of bone metastasis. This could help clinicians in the identification of patients who likely can develop bone metastasis and who could benefit from personalized treatments and follow-up strategies to prevent metastatic disease.


Pulmonary deposition sites of an inhaled radiolabeled submicronic aerosol.
The clearance rate of aerosolized and deposited 99mtechnetium diethylenetriamine pentacetate provides an index of lung epithelial permeability. However, the location of the epithelium being assessed is uncertain. We determined the percentage of submicronic aerosol deposited on ciliated and nonciliated airways in healthy and damaged lungs using 99mTc-sulphur (99mTc-S) colloid. Colloidal particles can be cleared by mucociliary transport but not through the epithelium. Piglets aged 12-72 h, weighing 0.7-3.3 kg (average 1.6 kg) were anesthetized and underwent endotracheal intubation. 99mTc-S labeled submicronic aerosol (Syntevent II) was collected in anesthetic bags and either inhaled spontaneously (n = 4), or insufflated by hand (n = 5) at 30 breaths per min and a peak airway pressure of 15 cm H2O. Piglets were immediately extubated and scanned on a gamma camera. Twenty-four h later a repeat scan was performed. The residual radioactivity represents the amount of aerosol deposited on nonciliated airways. In six other piglets pulmonary damage was produced by an intravenous infusion of air microemboli for 2 to 3 h (0.14 to 0.38 ml/min). The maximal decrease in PaO2 while breathing room air ranged from 24-59 mm Hg. During the last 15 min of infusion they were intubated, insufflated by hand with the 99mTc-S aerosol, and scanned as above. The results demonstrate that approximately three-fourths of the submicronic aerosol is deposited distally to ciliated airways in both healthy and damaged newborn piglet lungs. This suggests that the clearance rate of 99mtechnetium diethylenetriamine pentacetate predominantly reflects epithelial permeability of terminal lung units when a submicronic aerosol is used for delivery to the lung.


Roquin-2 promotes ubiquitin-mediated degradation of ASK1 to regulate stress responses.
Apoptosis signal-regulating kinase 1 (ASK1, also known as MAP3K5) mediates reactive oxygen species (ROS)-induced cell death. When activated by ROS, ASK1 ultimately becomes ubiquitinated and degraded by the proteasome, a process that is antagonized by the ubiquitin-specific protease USP9X. Using a functional siRNA (small interfering RNA) screen in HeLa cells, we identified Roquin-2 (also called RC3H2) as an E3 ubiquitin ligase required for ROS-induced ubiquitination and degradation of ASK1. In cells treated with H2O2, knockdown of Roquin-2 promoted sustained activation of ASK1 and the downstream stress-responsive kinases JNK (c-Jun amino-terminal kinase) and p38 MAPK (mitogen-activated protein kinase), and led to cell death. The nematode Caenorhabditis elegans produces ROS as a defense mechanism in response to bacterial infection. In C. elegans, mutation of the gene encoding the Roquin-2 ortholog RLE-1 promoted accumulation of the activated form of the ASK1 ortholog NSY-1 and conferred resistance to infection by the bacteria Pseudomonas aeruginosa. Thus, these data suggest that degradation of ASK1 mediated by Roquin-2 is an evolutionarily conserved mechanism required for the appropriate regulation of stress responses, including pathogen resistance and cell death.


Performance of the 2010 ACR/EULAR criteria for rheumatoid arthritis: comparison with 1987 ACR criteria in a very early synovitis cohort.
Early identification of patients with rheumatoid arthritis (RA) is essential to allow the prompt institution of therapy. The 2010 American College of Rheumatology (ACR)/European League Against Rheumatism (EULAR) classification criteria, which replace the 1987 classification criteria, have been developed to facilitate such identification in patients with newly presenting inflammatory arthritis. This study therefore assesses the performance of these new criteria in patients with early synovitis. Data were analysed from patients with synovitis seen within 3 months of the onset of inflammatory arthritis. Patients were followed for 18 months to determine outcomes, and data on the cumulative fulfilment of 2010 and 1987 criteria and therapy were recorded. 265 patients were included in the study. 60 had alternative diagnoses at baseline. Of the remaining 205 patients, 20% fulfilled both 1987 and 2010 criteria, 3% fulfilled only 1987 criteria and 22% fulfilled only 2010 criteria at baseline. The 2010 criteria, when applied at baseline, detected more patients who eventually required disease-modifying antirheumatic drugs (DMARD) (65 (62%) vs 40 (38%); p<0.001), especially methotrexate (50 (68%) vs 31 (42%); p<0.01), within the first 18 months. However, more patients whose disease eventually resolved without ever requiring DMARD were classified at baseline as RA according to the 2010 criteria than with the 1987 criteria (16 (8%) vs 5 (2%); p=0.01). The 2010 ACR/EULAR criteria allow more rapid identification of patients requiring methotrexate compared with the 1987 ACR criteria when applied at baseline. However, overdiagnosis is an important issue to consider if these criteria are to be used in very early disease.


The peritoneal osmotic conductance is low well before the diagnosis of encapsulating peritoneal sclerosis is made.
Encapsulating peritoneal sclerosis (EPS) is a serious condition whose frequency is increasing the longer the duration of peritoneal dialysis. To identify prognostic indicators of EPS, we studied here longitudinal changes in peritoneal membrane function of patients who later developed this complication. We identified all patients with an unequivocal diagnosis of EPS who began their peritoneal dialysis in our unit over a 20-year period and matched each of them for dialysis duration and age with four control patients who completed their dialysis. The dialysate/plasma creatinine ratio increased with time in both groups but was significantly higher in the patients with EPS only at the time their dialysis was discontinued. The ultrafiltration capacity was significantly worse for at least 2 years before stopping dialysis, diverging further at the time dialysis ceased, suggesting reduced osmotic conductance in the EPS patients. Both the glucose exposure rate for the 5 years preceding stoppage of dialysis and exposure to the osmotic agent icodextrin were significantly higher. Residual renal function was less in the EPS group, but there was no significant difference in the rates of peritonitis compared to the control group. The 24 h peritoneal protein clearance was not significantly different in EPS patients, possibly due to a greater fibrous matrix. Thus, our study shows that regular peritoneal membrane function tests can identify most patients at high risk of developing EPS before its occurrence.


Plant functional traits have globally consistent effects on competition.
Phenotypic traits and their associated trade-offs have been shown to have globally consistent effects on individual plant physiological functions, but how these effects scale up to influence competition, a key driver of community assembly in terrestrial vegetation, has remained unclear. Here we use growth data from more than 3 million trees in over 140,000 plots across the world to show how three key functional traits--wood density, specific leaf area and maximum height--consistently influence competitive interactions. Fast maximum growth of a species was correlated negatively with its wood density in all biomes, and positively with its specific leaf area in most biomes. Low wood density was also correlated with a low ability to tolerate competition and a low competitive effect on neighbours, while high specific leaf area was correlated with a low competitive effect. Thus, traits generate trade-offs between performance with competition versus performance without competition, a fundamental ingredient in the classical hypothesis that the coexistence of plant species is enabled via differentiation in their successional strategies. Competition within species was stronger than between species, but an increase in trait dissimilarity between species had little influence in weakening competition. No benefit of dissimilarity was detected for specific leaf area or wood density, and only a weak benefit for maximum height. Our trait-based approach to modelling competition makes generalization possible across the forest ecosystems of the world and their highly diverse species composition.


Identification and Characterization of Hypoxia-Regulated Endothelial Circular RNA.
Circular RNAs (circRNAs) are noncoding RNAs generated by back splicing. Back splicing has been considered a rare event, but recent studies suggest that circRNAs are widely expressed. However, the expression, regulation, and function of circRNAs in vascular cells is still unknown. Here, we characterize the expression, regulation, and function of circRNAs in endothelial cells. Endothelial circRNAs were identified by computational analysis of ribo-minus RNA generated from human umbilical venous endothelial cells cultured under normoxic or hypoxic conditions. Selected circRNAs were biochemically characterized, and we found that the majority of them lacks polyadenylation, is resistant to RNase R digestion and localized to the cytoplasm. We further validated the hypoxia-induced circRNAs cZNF292, cAFF1, and cDENND4C, as well as the downregulated cTHSD1 by reverse transcription polymerase chain reaction in cultured endothelial cells. Cloning of cZNF292 validated the predicted back splicing of exon 4 to a new alternative exon 1A. Silencing of cZNF292 inhibited cZNF292 expression and reduced tube formation and spheroid sprouting of endothelial cells in vitro. The expression of pre-mRNA or mRNA of the host gene was not affected by silencing of cZNF292. No validated microRNA-binding sites for cZNF292 were detected in Argonaute high-throughput sequencing of RNA isolated by cross-linking and immunoprecipitation data sets, suggesting that cZNF292 does not act as a microRNA sponge. We show that the majority of the selected endothelial circRNAs fulfill all criteria of bona fide circRNAs. The circRNA cZNF292 exhibits proangiogenic activities in vitro. These data suggest that endothelial circRNAs are regulated by hypoxia and have biological functions.


Contact-Angle Hysteresis and Contact-Line Friction on Slippery Liquid-like Surfaces.
Contact-line pinning and dynamic friction are fundamental forces that oppose the motion of droplets on solid surfaces. Everyday experience suggests that if a solid surface offers low contact-line pinning, it will also impart a relatively low dynamic friction to a moving droplet. Examples of such surfaces are superhydrophobic, slippery porous liquid-infused, and lubricant-impregnated surfaces. Here, however, we show that slippery omniphobic covalently attached liquid-like (SOCAL) surfaces have a remarkable combination of contact-angle hysteresis and contact-line friction properties, which lead to very low droplet pinning but high dynamic friction against the motion of droplets. We present experiments of the response of water droplets to changes in volume at controlled temperature and humidity conditions, which we separately compare to the predictions of a hydrodynamic model and a contact-line model based on molecular kinetic theory. Our results show that SOCAL surfaces offer very low contact-angle hysteresis, between 1 and 3°, but an unexpectedly high dynamic friction controlled by the contact line, where the typical relaxation time scale is on the order of seconds, 4 orders of magnitude larger than the prediction of the classical hydrodynamic model. Our results highlight the remarkable wettability of SOCAL surfaces and their potential application as low-pinning, slow droplet shedding surfaces.


The turning point and end of an expanding epidemic cannot be precisely forecast.
Epidemic spread is characterized by exponentially growing dynamics, which are intrinsically unpredictable. The time at which the growth in the number of infected individuals halts and starts decreasing cannot be calculated with certainty before the turning point is actually attained; neither can the end of the epidemic after the turning point. A susceptible-infected-removed (SIR) model with confinement (SCIR) illustrates how lockdown measures inhibit infection spread only above a threshold that we calculate. The existence of that threshold has major effects in predictability: A Bayesian fit to the COVID-19 pandemic in Spain shows that a slowdown in the number of newly infected individuals during the expansion phase allows one to infer neither the precise position of the maximum nor whether the measures taken will bring the propagation to the inhibition regime. There is a short horizon for reliable prediction, followed by a dispersion of the possible trajectories that grows extremely fast. The impossibility to predict in the midterm is not due to wrong or incomplete data, since it persists in error-free, synthetically produced datasets and does not necessarily improve by using larger datasets. Our study warns against precise forecasts of the evolution of epidemics based on mean-field, effective, or phenomenological models and supports that only probabilities of different outcomes can be confidently given.


Identification of natural dyes on laboratory-dyed wool and ancient wool, silk, and cotton fibers using attenuated total reflection (ATR) Fourier transform infrared (FT-IR) spectroscopy and Fourier transform Raman spectroscopy.
Attenuated total reflection (ATR) infrared and Fourier transform (FT) Raman spectra were obtained from wool threads dyed in the laboratory with natural dyes used in antiquity, following a procedure similar to ancient methods for dyeing wool. The ATR spectra were primarily dominated by the signals of the wool, making it difficult to identify the dye on the fibers only by visual inspection of the infrared spectrum. However, the Raman spectra showed more significant characteristics attributable to the dyes as previously studied in the literature on modern synthetic dyes. A library-search method was thus applied to the second derivatives of both the ATR and Raman spectra to verify the possibility of identifying the dye. Two libraries were constructed, one consisting of the ATR spectra of undyed wool (raw, washed, and mordanted) and the transmission spectra of pure dyes and the other consisting of the Raman spectra of undyed wool and of pure dyes. Correlation and first-derivative correlation search algorithms were used. The results presented here suggest that the two types of spectroscopy are complementary in this kind of work, allowing the almost complete identification of historic dyes on wool. In fact, through the combined use of the two searches, most dyes were identified with a good index of similarity and within the first five hits. Only for annatto was identification totally impossible using either technique. Subsequently the same method was applied to wool, silk, and cotton threads taken from ancient Caucasian and Chinese textiles.


New concepts of neural regulation in human nasal mucosa.
Nasal mucosa is innervated by multiple subsets of nociceptive, parasympathetic and sympathetic nerves. These play carefully coordinated roles in regulating glandular, vascular and other processes. These functions are vital for cleaning and humidifying ambient air before it is inhaled into the lungs. The recent recognition of distinct classes of nociceptive nerves with unique patterns of sensory receptors that include seven transmembrane G-protein coupled receptors, new families of transient receptor potential and voltage and calcium gated ion channels, and combinations of neurotransmitters that can be modulated during inflammation by neurotrophic factors has revolutionized our understanding of the complexity and subtlety of nasal innervation. These findings may provide a rational basis for responses to air temperature changes, culinary and botanical odorants ("aromatherapy"), and inhaled irritants in conditions as diverse as idiopathic nonallergic rhinitis, occupational rhinitis, hyposmia, and multiple chemical sensitivity.


Process Improvement Initiative for the Perioperative Management of Patients With a Cardiovascular Implantable Electronic Device.
Economic, personnel, and procedural challenges often complicate and interfere with efficient and safe perioperative care of patients with cardiovascular implantable electronic devices (CIEDs). In the context of a process improvement initiative, we created and implemented a comprehensive anesthesiologist-run perioperative CIED service to respond to all routine requests for perioperative CIED consultations at a large academic medical center. This study was designed to determine whether this new care model was associated with improved operating room efficiency, reduced institutional cost, and adequate patient safety. We included patients with a CIED and a concurrent cohort of patients with the same eligibility criteria but without a CIED who underwent first-case-of-the-day surgery during the periods between February 1, 2008, and August 17, 2010 (preintervention) and between March 4, 2012, and August 1, 2014 (postintervention). The primary end point was delay in first-case-of-the day start time. We used multiple linear regression to compare delays in start times during the preintervention and postintervention periods and to adjust for potential confounders. A patient safety database was queried for CIED-related complications. Cost analysis was based on labor minutes saved and was calculated using nationally published administrative estimates. A total of 18,148 first-case surgical procedures were performed in 15,100 patients (preintervention period-7293 patients and postintervention period-7807 patients). Of those, 151 (2.1%) patients had a CIED in the preintervention period, and 146 (1.9%) had a CIED in the postintervention period. After adjustment for imbalances in baseline characteristics (age, American Society of Anesthesiologists physical status, and surgical specialty), the difference in mean first-case start delay between the postintervention and preintervention periods in the cohort of patients with a CIED was -16.7 minutes (95% confidence interval [CI], -26.1 to -7.2). The difference in mean delay between the postintervention and preintervention periods in the cohort without a CIED was -4.7 minutes (95% CI, -5.4 to -3.9). There were 3 CIED-related adverse events during the preintervention period and none during the postintervention period. Based on reduction in first-case start delay, the intervention was associated with cost savings (estimated institutional savings $14,102 annually, or $94.06 per CIED patient), with a return on investment ratio of 2.18 over the course of the postintervention period. Based on our experience, specially trained anesthesiologists can provide efficient and safe perioperative care for patients with CIEDs. Other centers may consider implementing a similar strategy as our specialty adopts the perioperative surgical home model.


Herbal bathing: an analysis of variation in plant use among Saramaccan and Aucan Maroons in Suriname.
Herbal baths play an important role in the traditional health care of Maroons living in the interior of Suriname. However, little is known on the differences in plant ingredients used among and within the Maroon groups. We compared plant use in herbal baths documented for Saramaccan and Aucan Maroons, to see whether similarity in species was related to bath type, ethnic group, or geographical location. We hypothesized that because of their dissimilar cultural background, they used different species for the same type of bath. We assumed, however, that plants used in genital baths were more similar, as certain plant ingredients (e.g., essential oils), are preferred in these baths. We compiled a database from published and unpublished sources on herbal bath ingredients and constructed a presence/absence matrix per bath type and study site. To assess similarity in plant use among and within Saramaccan and Aucan communities, we performed three Detrended Correspondence Analyses on species level and the Jaccard Similarity Index to quantify similarity in bath ingredients. We recorded 349 plants used in six commonly used bath types: baby strength, adult strength, skin diseases, respiratory ailments, genital steam baths, and spiritual issues. Our results showed a large variation in plant ingredients among the Saramaccan and Aucans and little similarity between Saramaccans and Aucans, even for the same type of baths. Plant ingredients for baby baths and genital baths shared more species than the others. Even within the Saramaccan community, plant ingredients were stronger associated with location than with bath type. Plant use in bathing was strongly influenced by study site and then by ethnicity, but less by bath type. As Maroons escaped from different plantations and developed their ethnomedicinal practices in isolation, there has been little exchange in ethnobotanical knowledge after the seventeenth century between ethnic groups. Care should be taken in extrapolating plant use data collected from one location to a whole ethnic community. Maroon plant use deserves more scientific attention, especially now as there are indications that traditional knowledge is disappearing.


Drosophila NAP-1 is a core histone chaperone that functions in ATP-facilitated assembly of regularly spaced nucleosomal arrays.
We describe the cloning and analysis of Drosophila nucleosome assembly protein 1 (dNAP-1), a core histone-binding protein that functions with other chromatin assembly activities in a Drosophila chromatin assembly factor 1-containing fraction (dCAF-1 fraction) in the ATP-facilitated assembly of regularly spaced nucleosomal arrays from purified core histones and DNA. Purified, recombinant dNAP-1 acts cooperatively with a factor(s) in the dCAF-1 fraction in the efficient and DNA replication-independent assembly of chromatin. In the presence of histone H1, the repeat length of the chromatin is similar to that of native chromatin from Drosophila embryos. By coimmunoprecipitation analysis, dNAP-1 was found to be associated with histones H2A and H2B in a crude whole-embryo extract, which suggests that dNAP-1 is bound to the histones in vivo. Studies of the localization of dNAP-1 in the Drosophila embryo revealed that the factor is present in the nucleus during S phase and is predominantly cytoplasmic during G2 phase. These data suggest that NAP-1 acts as a core histone shuttle which delivers the histones from the cytoplasm to the chromatin assembly machinery in the nucleus. Thus, NAP-1 appears to be one component of a multifactor chromatin assembly machinery that mediates the ATP-facilitated assembly of regularly spaced nucleosomal arrays.


Abrupt and gradual extinction among Late Permian land vertebrates in the Karoo basin, South Africa.
The Karoo basin of South Africa exposes a succession of Upper Permian to Lower Triassic terrestrial strata containing abundant terrestrial vertebrate fossils. Paleomagnetic/magnetostratigraphic and carbon-isotope data allow sections to be correlated across the basin. With this stratigraphy, the vertebrate fossil data show a gradual extinction in the Upper Permian punctuated by an enhanced extinction pulse at the Permian-Triassic boundary interval, particularly among the dicynodont therapsids, coinciding with negative carbon-isotope anomalies.


Assessment of radiofrequency exposure from cellular telephone daily use in an epidemiological study: German Validation study of the international case-control study of cancers of the brain--INTERPHONE-Study.
The objective of the study is to validate self-reported cellular phone use information by comparing it with the cumulative emitted power and duration of calls measured by software-modified cellular phones (SMP). The information was obtained using a questionnaire developed for the international case-control study on the risk of the use of mobile phones in tumours of the brain or salivary gland (INTERPHONE-study). The study was conducted in Bielefeld, Germany. Volunteers were asked to use SMPs instead of their own cellular phones for a period of 1 month. The SMP recorded the power emitted by the mobile phone handset during each base station contact. Information on cellular phone use for the same time period from traffic records of the network providers and from face-to-face interviews with the participants 3 months after the SMP use was assessed. Pearson's correlation coefficients and linear regression models were used to analyse the association between information from the interview and from the SMP. In total, 1757 personal mobile phone calls were recorded for 45 persons by SMP and traffic records. The correlation between the self-reported information about the number and the duration of calls with the cumulative power of calls was 0.50 (P<0.01) and 0.48 (P<0.01), respectively. Almost 23% of the variance of the cumulative power was explained by either the number or the cumulative duration of calls. After inclusion of possible confounding factors in the regression model, the variance increased to 26%. Minor confounding factors were "network provider", "contract form", and "cellular phone model". The number of calls alone is a sufficient parameter to estimate the cumulative power emitted by the handset of a cellular telephone. The cumulative power emitted by these phones is only associated with number of calls but not with possible confounding factors. Using the mobile phone while driving, mainly in cities, or mainly in rural areas is not associated with the recorded cumulative power in the SMP.


In situ grafting made easy. Modification of a technique.
In situ bypass grafting has come of age following a period of disenchantment. It has become, in some centers, the operation of choice for bypasses to the tibial and peroneal vessels. Two fundamentally different techniques for this procedure have evolved. The first, following the method of Hall, uses a retrograde valvulotome passed from the distal saphenous vein to the proximal vein; on withdrawal, it severs the valves. A second approach, that of Leather, uses microscissors and a right-angle valvulotome, along with a disposable valve stripper, to disrupt the valves. Proponents exist for both approaches. In 1983 we reported our protocol for in situ grafting using the Hall technique. A number of modifications have since been made to expedite the technique. We describe and provide illustrations of these modifications.


Endoscopic excision of Juvenile nasopharyngeal angiofibroma - A comprehensive approach.
Endoscopie excision of Juvenile Nasopharyngeal Angiofibroma (JNA) was carried out with (he objective of minimizing blood loss and attempting a complete excision of the tumor under direct vision with the help of Hopkins telescopes. A prospective 4 year study of 23 cases of JNA treated by endoscopie excision is presented. Of these, 18 were treated by endoscopie excision alone. The remaining 5 were treated with a two staged approach either by mid-facial degloving followed by endoscopy or by 2 endoscopie procedures. The tumor was excised completely in 17 out of the total 18 cases that were treated exclusively by endoscopy. One case has shown a recurrence. The 5 cases treated by the staged approach represented very large tumours or tumours with intra-cranial extensions. In I of these cases, inoperable tumor remnant engulfing the internal carotid artery was treated by radiotherapy post-operatively. With successful excision of JNA in all but one case, we could reasonably conclude, that endoscopie excision of JNA could become a safer and a more precise alternative to open surgery provided it is practiced judiciously by surgeons who have considerable experience in endoscopie surgery and the necessary backup to convert to open surgery should the need arise.


Measurement of FNR-NrdI Interaction by Microscale Thermophoresis (MST).
This protocol describes how to measure protein-protein interactions by microscale thermophoresis (MST) using the MonolithTM NT.115 instrument (NanoTemper). We have used the protocol to determine the binding affinities between three different flavodoxin reductases (FNRs) and a flavodoxin-like protein, NrdI, from Bacillus cereus ( Lofstad et al., 2016 ). NrdI is essential in the activation of the manganese-bound form of the class Ib ribonucleotide reductase (RNR) system. RNRs, in turn, are the only source of the de novo synthesis of deoxyribonucleotides required for DNA replication and repair in all living organisms.


Positive Patient Response to a Structured Exercise Program Delivered in Inpatient Psychiatry.
The complexity of diagnostic presentations of an inpatient psychiatry population requires an integrative approach to health and well-being. In this sense, the primary need of this research aims at developing clinical strategies and healthier coping skills for anger, anxiety, and depression; promoting self-esteem, healthier sleep, and anxiety reduction; as well as enhancing mood and emotional-behavioral regulation via exercise and nutrition education. The primary objective is to promote exercise, fitness, and physical health in inpatient psychiatry patients. The secondary objective includes therapeutic management of depressive symptoms and patient-centered approach to mania, angry outbursts, and generalized disruptive behavior. The tertiary objective is promoting research in the psychophysiological effectors of exercise and nutrition education in combination with psychotherapy. Monitoring self-reported changes in mood and general well-being via administration of surveys and questionnaires pre- and postexercise sessions. The research yielded positive outcomes in all areas investigated, suggesting the positive effects of exercise and mind-body strategies in the context of psychotherapy in inpatient psychiatry. Physical exercise may be a helpful way to reduce mental health disorders in the context of inpatient psychiatry by targeting anxiety, depression, anger, psychomotor agitation, and muscle tension and addressing stressors and triggers and to develop a more balanced and integrated sense of self.


Changes in plasma ACTH levels and corticotroph tumor size in patients with Cushing's disease during long-term treatment with the glucocorticoid receptor antagonist mifepristone.
Pituitary effects of long-term therapy with mifepristone, a glucocorticoid receptor antagonist, in Cushing's disease (CD) patients are not well understood. Our objective was to report changes in ACTH and pituitary magnetic resonance imaging (MRI) findings during long-term use of mifepristone in CD patients. The Study of the Efficacy and Safety of Mifepristone in the Treatment of Endogenous Cushing's Syndrome (SEISMIC) was a 24-week, open-label study of mifepristone, and its long-term extension (LTE) is a multicenter U.S. study. Forty-three CD patients (mean age 45.3 years) were enrolled in SEISMIC with 27 continuing into the LTE study. Mifepristone (300-1200 mg) was administered once daily. ACTH and pituitary MRI were assessed at baseline and at regular intervals during treatment. A ≥2-fold increase in ACTH was observed in 72% of patients treated for a median duration of 11.3 months. The mean peak increase in ACTH was 2.76 ± 1.65-fold during SEISMIC, and mean ACTH concentrations remained stable during the LTE. ACTH was directly correlated with mifepristone dose and declined to near baseline levels after mifepristone discontinuation. Tumor regressed in 2 patients and progressed in 3 patients with macroadenomas. An additional microadenoma was identified after 25 months of treatment after a baseline tumor-negative MRI. In the largest prospective study to date, long-term mifepristone treatment increased ACTH in approximately two-thirds of patients with CD. ACTH elevations were observed within the first few weeks of treatment, were dose-dependent, and generally remained stable over time. Corticotroph tumor progression and regression may occur over time, but patients may have significant increases in ACTH levels without evidence of tumor growth.


β-secretase cleavage of the fly amyloid precursor protein is required for glial survival.
β-secretase (or BACE1) is the key enzyme in the production of β-amyloid (Aβ), which accumulates in the senile plaques characteristic for Alzheimer's disease. Consequently, the lack of BACE1 prevents β-processing of the amyloid precursor protein and Aβ production, which made it a promising target for drug development. However, the loss of BACE1 is also detrimental, leading to myelination defects and altered neuronal activity, functions that have been associated with the cleavage of Neuregulin and a voltage-gated sodium channel subunit. Here we show that the Drosophila ortholog of BACE, dBACE, is required for glial survival. Cell-specific knockdown experiments reveal that this is a non-cell autonomous function, as a knockdown of dBACE in photoreceptor neurons leads to progressive degeneration of glia in their target zone, the lamina. Interestingly, this phenotype is suppressed by the loss of the fly amyloid precursor protein (APPL), whereas a secretion-deficient form of APPL enhances the degeneration. This shows that full-length APPL in neurons promotes the death of neighboring glial cells and that β-processing of APPL is needed to prevent glial death. These results therefore not only demonstrate a novel function for an APP protein in glia, but they also show this function specifically requires regulation by β-cleavage.


Diverse definitions of prolonged labour and its consequences with sometimes subsequent inappropriate treatment.
Prolonged labour very often causes suffering from difficulties that may have lifelong implications. This study aimed to explore the prevalence and treatment of prolonged labour and to compare birth outcome and women's experiences of prolonged and normal labour. Women with spontaneous onset of labour, living in a Swedish county, were recruited two months after birth, to a cross-sectional study. Women (n = 829) completed a questionnaire that investigated socio-demographic and obstetric background, birth outcome and women's feelings and experiences of birth. The prevalence of prolonged labour, as defined by a documented ICD-code and inspection of partogram was calculated. Four groups were identified; women with prolonged labour as identified by documented ICD-codes or by partogram inspection but no ICD-code; women with normal labour augmented with oxytocin or not. Every fifth woman experienced a prolonged labour. The prevalence with the documented ICD-code was (13%) and without ICD-code but positive partogram was (8%). Seven percent of women with prolonged labour were not treated with oxytocin. Approximately one in three women (28%) received oxytocin augmentation despite having no evidence of prolonged labour. The length of labour differed between the four groups of women, from 7 to 23 hours.Women with a prolonged labour had a negative birth experience more often (13%) than did women who had a normal labour (3%) (P <0.00). The factors that contributed most strongly to a negative birth experience in women with prolonged labour were emergency Caesarean section (OR 9.0, 95% CI 1.2-3.0) and to strongly agree with the following statement 'My birth experience made me decide not to have any more children' (OR 41.3, 95% CI 4.9-349.6). The factors that contributed most strongly to a negative birth experience in women with normal labour were less agreement with the statement 'It was exiting to give birth' (OR 0.13, 95% CI 0.34-0.5). There is need for increased clinical skill in identification and classification of prolonged labour, in order to improve care for all women and their experiences of birthing processes regardless whether they experience a prolonged labour or not.


A narrative review of generic intervention fidelity measures.
To increase the rigor of pediatric rehabilitation research, there is a need to evaluate the degree to which an intervention is conducted as planned (i.e., fidelity). Generic fidelity measures evaluate more than one intervention and often include nonspecific attributes of the therapy process common to both interventions. The objective of this study was to describe the characteristics of generic fidelity measures and examine how these attributes fit with pediatric rehabilitation. A review of generic fidelity measures was conducted utilizing health and education databases. Five generic fidelity measures are described and examined for their applicability to pediatric rehabilitation. The measures were used in nine studies meeting the inclusion criteria, involving people ages from 11 years to >65 years undergoing psychotherapy. From the 76 nonspecific items, 37 items were judged to be applicable to pediatric rehabilitation. Common characteristics of nonspecific attributes with pediatric rehabilitation are discussed, and investigator plans to conduct future testing.


Meeting the methodological challenges in molecular mapping of the embryonic epigenome.
The past decade of life sciences research has been driven by progress in genomics. Many voices are already proclaiming the post-genomics era, in which phenomena other than sequence polymorphism influence gene expression and also explain complex phenotypes. One of these burgeoning fields is the study of the epigenome. Although the mechanisms by which chromatin structure and reorganization as well as cytosine methylation influence gene expression are not fully understood, they are being invoked to explain the now-accepted long-term impact of the environment on gene expression, which appears to be a factor in the development of numerous diseases. Such studies are particularly relevant in early embryonic development, during which waves of epigenetic reprogramming are known to have profound impacts. Since gametes and zygotes are in the process of resetting the genome in order to create embryonic stem cells that will each differentiate to create one of many specific tissue types, this phase of life is now viewed as a window of susceptibility to epigenetic reprogramming errors. Epigenetics could explain the influence of factors such as the nutritional/metabolic status of the mother or the artificial environment of assisted reproductive technologies. However, the peculiar nature of early embryos in addition to their scarcity poses numerous technological challenges that are slowly being overcome. The principal subject of this article is to review the suitability of various current and emerging technological platforms to study oocytes and early embryonic epigenome with more emphasis on studying DNA methylation. Furthermore, the constraint of samples size, inherent to the study of preimplantation embryo development, was put in perspective with the various molecular platforms described.


Design optimization of scaffold microstructures using wall shear stress criterion towards regulated flow-induced erosion.
Tissue scaffolds aim to provide a cell-friendly biomechanical environment for facilitating cell growth. Existing studies have shown significant demands for generating a certain level of wall shear stress (WSS) on scaffold microstructural surfaces for promoting cellular response and attachment efficacy. Recently, its role in shear-induced erosion of polymer scaffold has also drawn increasing attention. This paper proposes a bi-directional evolutionary structural optimization (BESO) approach for design of scaffold microstructure in terms of the WSS uniformity criterion, by downgrading highly-stressed solid elements into fluidic elements and/or upgrading lowly-stressed fluidic elements into solid elements. In addition to this, a computational model is presented to simulate shear-induced erosion process. The effective stiffness and permeability of initial and optimized scaffold microstructures are characterized by the finite element based homogenization technique to quantify the variations of mechanical properties of scaffold during erosion. The illustrative examples show that a uniform WSS is achieved within the optimized scaffold microstructures, and their architectural and biomechanical features are maintained for a longer lifetime during shear-induced erosion process. This study provides a mathematical means to the design optimization of cellular biomaterials in terms of the WSS criterion towards controllable shear-induced erosion.


Functional uncoupling of linked neurotransmitter effects by combinatorial convergence.
Physiological signaling pathways both diverge and converge-a single neurotransmitter can have multiple effects and multiple transmitters can have the same effects-in the same target cell. Divergence couples the effects of a transmitter together in a relatively fixed ratio. Different physiological circumstances may require a different ratio, however; the coupling must be made modifiable. This can be achieved through convergence. If two transmitters couple the effects in different ratios, then combinations of the transmitters can yield all intermediate ratios of the effects, thus functionally uncoupling them. This mechanism is analyzed in a well-understood, simple invertebrate neuromuscular circuit.


Dlk1 is a negative regulator of emerging hematopoietic stem and progenitor cells.
The first mouse adult-repopulating hematopoietic stem cells emerge in the aorta-gonad-mesonephros region at embryonic day (E) 10.5. Their numbers in this region increase thereafter and begin to decline at E12.5, thus pointing to the possible existence of both positive and negative regulators of emerging hematopoietic stem cells. Our recent expression analysis of the aorta-gonad-mesonephros region showed that the Delta-like homologue 1 (Dlk1) gene is up-regulated in the region of the aorta-gonad-mesonephros where hematopoietic stem cells are preferentially located. To analyze its function, we studied Dlk1 expression in wild-type and hematopoietic stem cell-deficient embryos and determined hematopoietic stem and progenitor cell activity in Dlk1 knockout and overexpressing mice. Its role in hematopoietic support was studied in co-culture experiments using stromal cell lines that express varying levels of Dlk1. We show here that Dlk1 is expressed in the smooth muscle layer of the dorsal aorta and the ventral sub-aortic mesenchyme, where its expression is dependent on the hematopoietic transcription factor Runx1. We further demonstrate that Dlk1 has a negative impact on hematopoietic stem and progenitor cell activity in the aorta-gonad-mesonephros region in vivo, which is recapitulated in co-cultures of hematopoietic stem cells on stromal cells that express varying levels of Dlk1. This negative effect of Dlk1 on hematopoietic stem and progenitor cell activity requires the membrane-bound form of the protein and cannot be recapitulated by soluble Dlk1. Together, these data suggest that Dlk1 expression by cells of the aorta-gonad-mesonephros hematopoietic microenvironment limits hematopoietic stem cell expansion and is, to our knowledge, the first description of such a negative regulator in this tissue.


A novel cell-free mitochondrial fusion assay amenable for high-throughput screenings of fusion modulators.
Mitochondria are highly dynamic organelles whose morphology and position within the cell is tightly coupled to metabolic function. There is a limited list of essential proteins that regulate mitochondrial morphology and the mechanisms that govern mitochondrial dynamics are poorly understood. However, recent evidence indicates that the core machinery that governs mitochondrial dynamics is linked within complex intracellular signalling cascades, including apoptotic pathways, cell cycle transitions and nuclear factor kappa B activation. Given the emerging importance of mitochondrial plasticity in cell signalling pathways and metabolism, it is essential that we develop tools to quantitatively analyse the processes of fission and fusion. In terms of mitochondrial fusion, the field currently relies upon on semi-quantitative assays which, even under optimal conditions, are labour-intensive, low-throughput and require complex imaging techniques. In order to overcome these technical limitations, we have developed a new, highly quantitative cell-free assay for mitochondrial fusion in mammalian cells. This assay system has allowed us to establish the energetic requirements for mitochondrial fusion. In addition, our data reveal a dependence on active protein phosphorylation for mitochondrial fusion, confirming emerging evidence that mitochondrial fusion is tightly integrated within the global cellular response to signaling events. Indeed, we have shown that cytosol derived from cells stimulated with different triggers either enhance or inhibit the cell-free fusion reaction. The adaptation of this system to high-throughput analysis will provide an unprecedented opportunity to identify and characterize novel regulatory factors. In addition, it provides a framework for a detailed mechanistic analysis of the process of mitochondrial fusion and the various axis of regulation that impinge upon this process in a wide range of cellular conditions.See Commentary: http://www.biomedcentral.com/1741-7007/8/99.


Procollagen type III N-terminal peptide (P3NP) and lean mass: a cross-sectional study.
Procollagen type III N-terminal peptide (P3NP) is released during collagen synthesis in muscle. Increased circulating P3NP is a marker not only of muscle growth, but also of muscle repair and fibrosis. Thus, P3NP may be a potential biomarker for sarcopenia. To determine the association between plasma P3NP and lean mass and strength. A cross-sectional study of men and women from the Framingham Offspring Study. Participants included a convenience sample of 687 members with a measure of plasma P3NP and lean mass, and 806 members with P3NP and quadriceps strength assessment. Linear regression was used to estimate the association between total and appendicular lean mass and plasma P3NP, and quadriceps strength and P3NP. Mean age was 58 years. Median plasma P3NP was similar in men (3.4 mg/L), premenopausal women (3.1 mg/L), and postmenopausal women (3.0 mg/L). In adjusted models, higher P3NP was associated with a modest decrease in total and appendicular lean mass in postmenopausal women [β= -0.13 unit P3NP/kg total lean mass; p=0.003]. A similar trend was found among premenopausal women, although results were not statistically significant [β=-0.10 unit P3NP/kg total lean mass; p=0.41]. No association between P3NP and lean mass was observed in men. P3NP was not associated with strength in men or women. Our results suggest that plasma P3NP might be a useful biomarker of muscle mass in postmenopausal women if longitudinal studies demonstrate that it has adequate sensitivity and specificity to predict muscle loss.


Quantifying the relative importance to patients of avoiding symptoms and outcomes of heart failure.
To evaluate heart failure (HF) patients' disease knowledge and preferences for avoiding different disease outcomes. An online survey was administered to 400 individuals with a self-reported diagnosis of HF to elicit relative importance weights (RIWs) for avoiding 11 potential HF symptoms and outcomes using best-worst scaling. The survey also included questions about individuals' HF knowledge, and demographic and disease-experience characteristics. Differences in RIWs among sub-groups, defined by HF knowledge, caregiver support, age, recent hospitalization or emergency room visit for HF, health-related quality-of-life, and cardiac device experience were examined. Relative to limitations in usual activities (RIW 1.00), respondents preferred avoiding severe, infrequent cardiovascular events (e.g. stroke [RIW 8.51], heart transplant [RIW 7.84], or heart attack [RIW 5.3]) most, followed by difficulty breathing (RIW 2.55), inability to enjoy life (RIW 1.84), cardiac device implantation (RIW 1.74), and atrial fibrillation (RIW 1.57). Patients preferred avoiding swelling (RIW 0.47) and fatigue (RIW 0.58) least. RIWs for avoiding severe, infrequent events were higher among those with high disease knowledge, those without caregivers, and those without a recent hospitalization or emergency room visit. Patients' preferences for avoiding HF outcomes vary across outcomes and by individuals' knowledge, caregiver status, and age. Healthcare providers should solicit and incorporate insights about patients' knowledge of HF and their preferences for avoiding HF outcomes into HF education and management planning efforts.