Scaphoid Nonunion.
The natural history and treatment of scaphoid fractures and subsequent nonunions have occupied a substantial portion of the orthopaedic literature. The authors examine the role of modern diagnostic tools in making an earlier diagnosis of scaphoid nonunion, in more accurately determining the displacement and angulation of the fragments, and in identifying the presence of avascular necrosis. They also consider the various available treatment modalities, including immobilization, electrical stimulation, both conventional and vascularized bone grafting, and internal fixation. Finally, a brief review of salvage procedures and the authors' preferred treatment are presented.


Needs of caregivers in heart failure management: A qualitative study.
To identify the needs of caregivers supporting a person with heart failure and to inform the development of a caregiver resource to be used as part of a home-based self-management programme. A qualitative study informed by thematic analysis involving 26 caregivers in individual interviews or a focus group. Three distinct aspects of caregiver support in heart failure management were identified. Firstly, caregivers identified needs about supporting management of heart failure including: coping with the variability of heart failure symptoms, what to do in an emergency, understanding and managing medicines, providing emotional support, promoting exercise and physical activity, providing personal care, living with a cardiac device and supporting depression management. Secondly, as they make the transition to becoming a caregiver, they need to develop skills to undertake difficult discussions about the role; communicate with health professionals; manage their own mental health, well-being and sleep; and manage home and work. Thirdly, caregivers require skills to engage social support, and voluntary and formal services while recognising that the long-term future is uncertain. The identification of the needs of caregiver has been used to inform the development of a home-based heart failure intervention facilitated by a trained health care practitioner.


The role of human immunodeficiency virus in the pathogenesis of vernal keratoconjunctivitis-like disease in adults: A demographic and epidemiological study.
The purpose was to identify and describe patients with new-onset vernal keratoconjunctivitis-like (VKC-like) disease after puberty. The study consisted of two parts: a prospective observational descriptive study of patients with new-onset VKC-like disease, and a case-control study to determine the relationship of a CD4 count with VKC-like disease in adults, in the setting of human immunodeficiency virus (HIV). Patients were recruited between January 2016 and November 2017 from a Provincial Eye hospital, one of two large referral hospitals in KwaZulu-Natal, South Africa. Patients presenting to the eye clinic were screened and diagnosed at the Primary Eye Care Unit. Inclusion criteria: age 15 years and older with signs and symptoms of new-onset VKC-like disease. Exclusion criteria: a history of childhood atopic diseases, atopic keratoconjunctivitis and patients who declined HIV testing. Data collected included HIV status, CD4 count, antinuclear antibodies and total serum immunoglobulin E. Thirty-three patients were included; females n = 16 and males n = 17. The mean age at presentation was 32.45 ± 9.93 years, 95% CI = 28.94-35.97. Twenty-six patients (78.8%) were HIV positive, 95% CI (62-89). The proportion of HIV positive patients was statistically different from the HIV negative group, Chi-squared = 21.866, P value <0.0001. In the group of HIV positive patients, 72% were classified as immunodeficient according to their CD4 counts. An association was proven between severely immunodeficient patients and the risk of VKC-like disease (Chi-squared = 4.992, P value = 0.0255). In this cohort, a statistically significant association was found between VKC-like disease in adults and an HIV positive status. This association calls for more research on the subject.


Root Rot and Dieback of Pinus pinea Caused by Phytophthora humicola in Tuscany, Central Italy.
High mortality was noticed in a 10-year-old stand of Pinus pinea in the Alberese area (Grosseto, central Italy, elev. 40 m, 42° 39' 46″ N, 11° 06' 25″ E) in July 2010. Aerial symptoms of trees included chlorosis, crown thinning, stunted growth, bark lesions at the stem base with resinous exudations, and extensive necroses of the underlying xylem tissue. Woody roots of two uprooted trees exhibited bark necroses and a high proportion of fine roots was destroyed. Soil around necrotic roots was baited using apple fruits (cv. Gala). After 1 week of incubation at 24°C, typical firm fruit rot developed and small tissue samples were transferred to clarified V8 agar (V8A) amended with 5 ml/l PARPNH and incubated at 24°C. After 7 days, stellate to rosaceous, finely lobed cottony colonies arose that were transferred to FPM medium and incubated at 24°C. Within 7 days, spherical oogonia with a smooth surface and predominantly paragynous antheridia formed; sporadic amphyginous antheridia could be observed. Colony squares (1 cm2) were then placed in filtered and sterilized pond water. After 48 h, ovoid, obpyriform, or clavate, nonpapillate, persistent sporangia with internal nested and extended proliferation were formed. Fifty oogonia and 30 sporangia were measured. The diameter of the 50 spherical oogonia varied from 33.6 to 44.9 μm (avg. 39 μm); dimensions of the 30 sporangia were 42.6 to 59.8 × 28.9 to 47.8 μm (avg. 52.95 × 38.98 μm; 1:b ratio 1.37). The isolate was identified as Phytophthora humicola W. H. Ko & Ann on the basis of colony type, size and morphology of oogonia and sporangia, average length/width ratio of sporangia, the homothallic formation of oogonia (4), and ITS rDNA sequence information (GenBank Accession No. JQ757060). A BLAST search of the ITS sequence of P. humicola isolate B33 revealed a 99% identity with the Phytophthora ITS Clade 6 species P. humicola and P. inundata (2). This latter species could be ruled out, however, since it is self-sterile, whereas our isolate B33 was self-fertile (3). A strain of P. humicola was deposited in the CBS-KNAW Fungal Biodiversity Centre, strain no. CBS129249. Pathogenicity tests were conducted on 10 one-year-old twigs of Pinus pinea. A bark portion was removed aseptically and a V8A disc (0.5 cm diam.) of P. humicola mycelium was placed on the wound. Control twigs (3) received sterile V8A discs. Inoculated and control twigs were incubated at 20°C in the dark. Clearly noticeable necrotic lesions (avg. length 2.2 × 0.68 cm) were observed after 15 days on inoculated twigs. Control twigs showed no symptoms. Reisolations on selective V8-PARPNH-agar confirmed P. humicola as the causal agent. P. humicola is mainly associated with woody horticultural crops (1, 3), while the other taxa grouped with this species in Clade 6 are mainly found in forest and riparian ecosystems (1). These aquatic Phytophthora species normally have a saprophytic lifestyle, but under favourable environmental circumstances can act as opportunistic pathogens, attacking susceptible trees and causing scattered mortality in forest stands and natural ecosystems (3). To our knowledge, this is the first report of P. humicola from a pine stand. It is supposed that the pathogen reached the stand through infected plant material or infested soil introduced into the stand. References: (1) C. M. Brasier et al. Mycol. Res. 107:277, 2003. (2) D. E. L. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (3) T. Jung et al. Persoonia 26:13, 2011. (4) W. H. Ko and P. J. Ann. Mycologia 77:631, 1985.


The Role of Biomarkers for the Diagnosis of Implant-Related Infections in Orthopaedics and Trauma.
Diagnosis of implant-related (periprosthetic joint) infections poses a major challenge to infection disease physicians and orthopaedic surgeons. Conventional diagnostic tests continue to suffer from issues of accuracy and feasibility. Biomarkers are used throughout medicine for diagnostic and prognostic purposes, as they are able to objectively determine the presence of a disease or a biological state. There is increasing evidence to support the measurement of specific biomarkers in serum and/or synovial fluid of patients with suspected periprosthetic joint infections. Promising serum biomarkers include interleukin (IL)-4, IL-6, tumour necrosis factor (TNF)-α, procalcitonin, soluble intercellular adhesion molecule 1 (sICAM-1), and D-dimer. In addition to c-reactive protein and leucocyte esterase, promising biomarkers that can be measured in synovial fluid include antimicrobial proteins such as human β-defensin (HBD)-2 and human β-defensin (HBD)-3, and cathelicidin LL-37, as well as several interleukins such as IL-1β, IL-6, IL-8, IL-17, TNF- α, interferon-δ, and vascular endothelial growth factor.


Adaptive particle swarm optimization.
An adaptive particle swarm optimization (APSO) that features better search efficiency than classical particle swarm optimization (PSO) is presented. More importantly, it can perform a global search over the entire search space with faster convergence speed. The APSO consists of two main steps. First, by evaluating the population distribution and particle fitness, a real-time evolutionary state estimation procedure is performed to identify one of the following four defined evolutionary states, including exploration, exploitation, convergence, and jumping out in each generation. It enables the automatic control of inertia weight, acceleration coefficients, and other algorithmic parameters at run time to improve the search efficiency and convergence speed. Then, an elitist learning strategy is performed when the evolutionary state is classified as convergence state. The strategy will act on the globally best particle to jump out of the likely local optima. The APSO has comprehensively been evaluated on 12 unimodal and multimodal benchmark functions. The effects of parameter adaptation and elitist learning will be studied. Results show that APSO substantially enhances the performance of the PSO paradigm in terms of convergence speed, global optimality, solution accuracy, and algorithm reliability. As APSO introduces two new parameters to the PSO paradigm only, it does not introduce an additional design or implementation complexity.


Synthesis, morphological control, and antibacterial properties of hollow/solid Ag2S/Ag heterodimers.
Ag(2)S and Ag are important functional materials that have received considerable research interest in recent years. In this work, we develop a solution-based synthetic method to combine these two materials into hollow/solid Ag(2)S/Ag heterodimers at room temperature. Starting from monodisperse Cu(2)O solid spheres, CuS hollow spheres can be converted from Cu(2)O through a modified Kirkendall process, and the obtained CuS can then be used as a solid precursor for preparation of the Ag(2)S/Ag heterodimers through ion exchange and photo-assisted reduction. We have found that formation of the Ag(2)S/Ag heterodimers is instantaneous, and the size of Ag nanocrystals on the hollow spheres of Ag(2)S can be controlled by changing the concentration and power of reducing agents in the synthesis. The growth of Ag nanoparticles on hollow spheres of Ag(2)S in the dimers is along the [111] direction of the silver crystal; the light absorption properties have also been investigated. Furthermore, coupling or tripling of Ag(2)S/Ag heterodimers into dumbbell-like trimers ((Ag(2)S)(2)/Ag, linear) and triangular tetramers ((Ag(2)S)(3)/Ag, coplanar) can also be attained at 60 degrees C by adding the bidentate ligand ethylenediamine as a cross-linking agent. To test the applicability of this highly asymmetric dipolar composite, photocatalytic inactivation of Escherichia coli K-12 in the presence of the as-prepared Ag(2)S/Ag heterodimers has been carried out under UV irradiation. The added Ag(2)S/Ag heterodimers show good chemical stability under prolonged UV irradiation, and no appreciable solid dissolution is found. Possible mechanisms regarding the enhanced antibacterial activity have also been addressed.


Quantitative nonisotopic nitrocellulose filter binding assays: bacterial manganese superoxide dismutase-DNA interactions.
Nitrocellulose filter binding assays (NCFBAs) have been used for many years to qualitatively and quantitatively determine protein-nucleic acid affinities. While this technique can be robust thermodynamically and fairly simple to perform, the requirement of radiolabeled nucleic acids (typically (32)P) has several major drawbacks. Some disadvantages are the short half-life of (32)P, the inherent safety concerns, and the cost of working with radioisotopes. Another drawback is that over time the beta emissions cause fragmentation of the nucleic acids. We have modified standard NCFBAs by developing a quantitative nonisotopic chemiluminescent method using biotin-labeled DNA and a dual-filter format. The biotin tag is detected on both nitrocellulose and positively charged nylon membranes by conventional streptavidin-conjugated alkaline phosphatase recognition systems. Quantitation of the photon emissions is simplified by use of a cooled charge-coupled device camera, although exposure to X-ray film and quantitation by densitometry could also be employed. The binding affinity of bacterial manganese superoxide dismutase to nonsequence-specific DNA has been quantitated previously by standard NCFBAs. We have replicated parts of a published binding study using identical solution conditions and the nonisotopic method that we have developed. We provide quantitative agreement between the isotopic and the nonisotopic methods.


Embodied energy and economic valuation.
Input-output analysis has been adapted to calculate the total (direct plus indirect) energy required to produce goods and services in the U.S. economy; this quantity has been termed the embodied energy. Usually, the energy required to produce labor and government services and the solar energy input to the economy are ignored by analysts. The former omission can be traced to the assumption that traditional primary factors of economic production-land, labor, and capital-are independent. A strong case can be made that these input factors are not independent and that energy is required for their production. Embodied energies can be calculated in this case by using input-output data. The results of such an analysis show that there is a strong relation between embodied energy and dollar value for a 92-sector U.S. economy if the energy required to produce labor and government services is included.


miR-543 and miR-590-3p regulate human mesenchymal stem cell aging via direct targeting of AIMP3/p18.
Previously, AIMP3 (aminoacyl-tRNAsynthetase-interacting multifunctional protein-3) was shown to be involved in the macromolecular tRNA synthetase complex or to act as a tumor suppressor. In this study, we report a novel role of AIMP3/p18 in the cellular aging of human mesenchymal stem cells (hMSCs). We found that AIMP3/p18 expression significantly increased in senescent hMSCs and in aged mouse bone marrow-derived MSCs (mBM-MSCs). AIMP3/p18 overexpression is sufficient to induce the cellular senescence phenotypes with compromised clonogenicity and adipogenic differentiation potential. To identify the upstream regulators of AIMP3/p18 during senescence, we screened for potential epigenetic regulators and for miRNAs. We found that the levels of miR-543 and miR-590-3p significantly decreased under senescence-inducing conditions, whereas the AIMP3/p18 protein levels increased. We demonstrate for the first time that miR-543 and miR-590-3p are able to decrease AIMP3/p18 expression levels through direct binding to the AIMP/p18 transcripts, which further compromised the induction of the senescence phenotype. Taken together, our data demonstrate that AIMP3/p18 regulates cellular aging in hMSCs possibly through miR-543 and miR-590-3p.


Pan-European longitudinal surveillance of antibiotic resistance among prevalent Clostridium difficile ribotypes.
Clostridium difficile infection remains a major healthcare burden. Until the recent introduction of fidaxomicin, antimicrobial treatments were limited to metronidazole and vancomycin. The emergence of epidemic C. difficile PCR ribotype 027 and its potential link to decreased antibiotic susceptibility highlight the lack of large-scale antimicrobial susceptibility and epidemiological data available. We report results of epidemiological and antimicrobial susceptibility investigations of C. difficile isolates collected prior to fidaxomicin introduction, establishing important baseline data. Thirty-nine sites in 22 countries submitted a total of 953 C. difficile isolates for PCR ribotyping, toxin testing, and susceptibility testing to metronidazole, vancomycin, fidaxomicin, rifampicin, moxifloxacin, clindamycin, imipenem, chloramphenicol, and tigecycline. Ninety-nine known ribotypes were identified. Ribotypes 027, 014, 001/072, and 078 were most frequently isolated in line with previous European studies. There was no evidence of resistance to fidaxomicin, and reduced susceptibility to metronidazole and vancomycin was also scarce. Rifampicin, moxifloxacin, and clindamycin resistance (13%, 40%, and 50% of total isolates, respectively) were evident in multiple ribotypes. There was a significant correlation between lack of ribotype diversity and greater antimicrobial resistance (measured by cumulative resistance score). Well-known epidemic ribotypes 027 and 001/072 were associated with multiple antimicrobial resistance, but high levels of resistance were also observed, particularly in 018 and closely related emergent ribotype 356 in Italy. This raises the possibility of antimicrobial exposure as the underlying reason for their appearance, and highlights the need for ongoing epidemiological and antimicrobial resistance surveillance.


Hereditary Non-Polyposis Colorectal Cancer: the rise and fall of a confusing term.
The term Hereditary Non-Polyposis Colorectal Cancer (HNPCC) is a poor descriptor of the syndrome described by Lynch. Over the last decade, the term has been applied to heterogeneous groups of families meeting limited clinical criteria, for example the Amsterdam criteria. It is now apparent that not all Amsterdam criteria-positive families have the Lynch syndrome. The term HNPCC has also been applied to clinical scenarios in which CRCs with DNA microsatellite instability are diagnosed but in which there is no vertical transmission of an altered DNA mismatch repair (MMR) gene. A term that has multiple, mutually incompatible meanings is highly problematic, particularly when it may influence the management of an individual family. The Lynch syndrome is best understood as a hereditary predisposition to malignancy that is explained by a germline mutation in a DNA MMR gene. The diagnosis does not depend in an absolute sense on any particular family pedigree structure or age of onset of malignancy. Families with a strong family history of colorectal cancer that do not have Lynch syndrome have been grouped as 'Familial Colorectal Cancer Type-X'. The first step in characterizing these cancer families is to distinguish them from Lynch syndrome. The term HNPCC no longer serves any useful purpose and should be phased out.


Enzymatically activated near infrared nanoprobes based on amphiphilic block copolymers for optical detection of cancer.
Nanotechnology offers the possibility of creating multi-functional structures that can provide solutions for biomedical problems. The nanoprobes herein described are an example of such structures, where nano-scaled particles have been designed to provide high specificity and contrast potential for optical detection of cancer. Specifically, enzymatically activated fluorescent nanoprobes (EANPs) were synthesized as cancer-specific contrast agents for optical imaging. EANPs were prepared by nanoprecipitation of blends of poly(lactic acid)-b-poly(ethylene glycol) and poly(lactic-co-glycolic acid)-b-poly(l-lysine). The lysine moieties were then covalently decorated with the near infrared (NIR) fluorescent molecule AlexaFluor-750 (AF750). Close proximity of the fluorescent molecules to each other resulted in fluorescence quenching, which was reversed by enzymatically mediated cleavage of poly(l-lysine) chains. EANPs were characterized by dynamic light scattering and electron microscopy. Enzymatic development of fluorescence was studied in vitro by fluorescence spectroscopy. Biocompatibility and contrast potential of EANPs were studied in cancerous and noncancerous cells. The potential of the nanoprobes as contrast agents for NIR fluorescence imaging was studied in tissue phantoms. Spherical EANPs of ∼100 nm were synthesized via nanoprecipitation of polymer blends. Fluorescence activation of EANPs by treatment with a model protease was demonstrated with up to 15-fold optical signal enhancement within 120 minutes. Studies with MDA-MB-231 breast cancer cells demonstrated the cytocompatibility of EANPs, as well as enhanced fluorescence associated with enzymatic activation. Imaging studies in tissue phantoms confirmed the ability of a simple imaging system based on a laser source and CCD camera to image dilute suspensions of the nanoprobe at depths of up to 4 mm, as well as up to a 13-fold signal-to-background ratio for enzymatically activated EANPs compared to un-activated EANPs at the same concentration. Nanoprecipitation of copolymer blends containing poly(l-lysine) was utilized as a method for preparation of highly functional nanoprobes with high potential as contrast agents for fluorescence based imaging of cancer. Lasers Surg. Med. 47:579-594, 2015. © 2015 Wiley Periodicals, Inc.


Suppression of glucose absorption by some fractions extracted from Gymnema sylvestre leaves.
Extracts containing gymnemic acids, which were extracted from the leaves of Gymnema sylvestre (GS) as nine fractions, were evaluated for their effects on a high K(+)-induced contraction of guinea-pig ileal longitudinal muscles, on glucose transport mediated by the difference of glucose-evoked transmural potential difference (delta PD) in the inverted intestine of guinea-pig and rat, and on blood glucose in rat. Among nine fractions obtained by high performance liquid chromatography from the extract, f-2 and f-4 strongly suppressed the high K(+)-induced contraction of the ileal muscle, f-3 and f-5 did so moderately, and f-8 and f-9 did so weakly, whereas the other fractions did not affect it. The degree of suppression of high K(+)-induced contraction by f-2 at 74% was almost the same as that of f-4 at 67%, at concentrations of 0.1 mg/ml. The suppressed contraction by f-2 or f-4 was recovered by adding 5.5 mM pyruvate. The delta PD increased by 5.5 mM glucose in the inverted intestines of guinea-pig and rat were equally suppressed by 0.1 mg/ml of f-2 or f-4 to 40%. In a rat sucrose tolerance test, f-2 and f-4 suppressed the elevation of blood glucose level. Both f-2 and f-4 suppressed the contraction of guinea-pig ileal longitudinal muscle, interfered with the increase in delta PD induced by glucose in the inverted intestines of guinea-pig and rat, and inhibited the elevation of blood glucose level. In conclusion, it is suggested that some of the extracts containing gymnemic acids from GS leaves suppress the elevation of blood glucose level by inhibiting glucose uptake in the intestine.


Hologram quantitative structure-activity relationships investigations of non-nucleoside reverse transcriptase inhibitors.
Non-nucleoside reverse transcriptase inhibitors (NNRTIs) such as TIBO, HEPT and dipyridodiazepinone are effective against HIV-1 RT. These NNRTIs are chemically and structurally diverse, but they all bind to a common allosteric site of HIV-1 RT. These inhibitors exhibit high potency, low cytotoxicity and produce few side effects. However, the emergency of drug-resistance viral strain has limited the therapeutic efficiency of the NNRTIs. Several different QSAR studies were reported to identify important structural features responsible for the inhibitory activity of these NNRTIs. In this study, hologram quantitative structure-activity relationships (HQSAR) was applied to three different data sets, 70 TIBO, 101 HEPT and 125 dipyridodiazepinone derivatives. Starting geometries of compounds were taken from available X-ray crystallographic data. Modification and full geometry optimization of all derivatives were performed, based on quantum chemical calculations at the HF/3-21G level of theory. All derived HQSAR models produce satisfying predictive ability and yield r(2)(cv) values ranging from 0.62-0.84. Moreover, it was also found that the quality of models enhances as the size of fragments increases. The obtained HQSAR results indicate the similarity of the interactions of these three different NNRTIs with the inhibition pocket of the enzyme. Comparisons of different QSAR methods on these NNRTIs data sets were also considered and it could be shown that HQSAR results yield superior predictive models than other 2D-QSAR approaches. In particular, the predictive ability of the models derived from dipyridodiazepinone analogues was significantly improved and apparently revealed differentiating structural requirements between WT and Y181C HIV RT inhibition. Additionally, the quality of QSAR models constructed by CoMFA and HQSAR methods are comparable and the interpretations of the models reinforce each other. It suggests an advantage of HQSAR as a useful tool in designing new potent inhibitors with enhanced HIV-1 RT inhibition activity, especially against mutant enzyme.


Molecular epidemiology of HIV-1 infection and full-length genomic analysis of circulating recombinant form 07_BC strains from injection drug users in Taiwan.
Previously, we reported that there was an outbreak of human immunodeficiency virus type 1 (HIV-1) circulating recombinant form (CRF) 07_BC among injection drug users (IDUs) in Taiwan in 2004. The objectives of the present study were to conduct a molecular epidemiological analysis and to characterize the full-length genome of the Taiwanese CRF07_BC. Three hundred and fifty-eight patients with HIV-1/AIDS from hospitals and 133 HIV-1-infected inmates from detention centers were recruited. DNA sequencing and phylogenetic analysis were conducted to determine subtypes and evolutionary relationships. Recombination breakpoints of 2 full-length CRF07_BC strains were elucidated using a bootscanning method. Of 206 HIV-1-infected patients who received a diagnosis in 2004, 44.7% were infected with subtype B, 53.4% with CRF07_BC, and 1.5% with CRF01_AE. Ninety-eight percent (109/111) of IDUs were infected with CRF07_BC. Deletions of 7-11 amino acids in both p6(gag) and p6(pol) proteins were noted among the Taiwanese CRF07_BC strains. The CRF_07BC strains belonged to 2 phylogenetic clusters, and the first cluster contained only CRF07_BC strains from the southern part of Taiwan. The Taiwanese CRF07_BC strains had 97% full-length sequence homology with the prototype from mainland China. CRF07_BC was first introduced into the southern region in 2002 and then spread to other regions in Taiwan in 2004.


[Explanations for the relation between depression and increased mortality].
Depression is a highly prevalent disorder at all levels of health-care delivery. Depression has an unfavourable effect on the prognosis of somatic illnesses, and is associated with excess mortality. Several mechanisms may contribute to these relationships. First of all, depression affects behaviour. Depressed patients show more unhealthy living habits, less compliance with medical treatment, and a higher number of accidents and suicides. Secondly, biological aspects of depression are important. Dysregulation of the neuro-immune system, hyperactivity of the hypothalamic pituitary adrenal axis, and autonomic dysregulation may all have a negative effect on both the prognosis of somatic illnesses and the lifespan. Although studies have suggested beneficial effects of the treatment of depression in (somatic) patients, it remains unclear whether treatment may also affect survival. Further research is needed to investigate the occurrence of and to unravel the mutual influences of depression and somatic illnesses, to search for possible pathogenetic mechanisms that may underlie both depression and medical disorders, and to assess the effects of depression treatment on biological parameters and survival.


Multicenter evaluation of a commercial PCR-enzyme-linked immunosorbent assay diagnostic kit (Onychodiag) for diagnosis of dermatophytic onychomycosis.
We prospectively evaluated a new PCR-enzyme-linked immunosorbent assay kit (Onychodiag; BioAdvance, France) for the diagnosis of dermatophytic onychomycosis by testing nail samples from 438 patients with suspected onychomycosis and from 108 healthy controls in three independent laboratories. In two laboratories, samples were collected by trained mycologists as close as possible to the lesions (proximal samples). In one laboratory, samples were collected by other physicians. All samples were processed by conventional mycological techniques and by Onychodiag, blindly to the mycological results. An additional distal sample, collected by clipping the nail plate, was obtained from 75 patients and tested with Onychodiag alone. In patients with culture-proven dermatophytic onychomycosis, the sensitivity of Onychodiag was 83.6% (87.9% including the gray zone) and ranged from 75 to 100% according to the laboratory and the sampling conditions. The specificity was 100% when healthy subjects were considered true negative controls. Onychodiag was positive on 68 patient samples that were sterile or yielded nondermatophyte species in culture. Based on the results of Onychodiag for mycologically proven positive samples and true-negative samples, these results were considered true positives, and the poor performance of mycology on these samples was attributed to inconvenient sampling conditions or to contaminants. When tested on distal samples, Onychodiag was positive in 49/53 (92%) cases of proven dermatophytic onychomycosis. Finally, with either proximal or distal samples, Onychodiag provided a diagnosis of dermatophytic onychomycosis within 24 to 48 h after sampling, and its sensitivity was close to that of mycological techniques applied to proximal samples.


Routine chest radiography after permanent pacemaker implantation: is it necessary?
Chest radiographs (CXRs) are performed routinely after permanent pacemaker implantation to identify pacemaker lead position and exclude pneumothorax. We assessed the clinical value and need for this procedure. Retrospective analysis of pacemaker data and CXRs following permanent pacemaker insertion between December 2002 and February 2004. Post-procedural CXRs were available in 125/126 consecutive patients after either first endocardial pacemaker implantation or insertion of at least one new lead. Subclavian vein puncture was used for venous access in all cases. CXRs were examined to establish the incidence of pneumothorax and assess pacing lead positions. The clinical records were examined in all patients who had subsequent CXRs or a further pacemaker procedure to identify the indication for these and to establish whether CXR had influenced patient management. In total, 192 post-procedural CXRs were performed, either postero-anterior (PA) and/or lateral views. Ventricular and/or atrial pacing lead contour and electrode position was considered radiographically appropriate in 86% CXRs. Fourteen per cent of post-procedural radiographs were considered to have radiologically sub-optimal pacemaker lead positioning. None of the patients with these "abnormal" radiographs experienced subsequent pacemaker complications or had further radiographs recorded at a later date. Later repeat CXRs were performed in 16 patients (13%) but only 3 patients (2%) had pacing abnormalities as the primary indication. All three had satisfactory pacing lead position on initial post-implantation and later radiographs, but required further procedures for lead re-positioning. Iatrogenic pneumothorax occurred in one patient (incidence 0.8%) in our series. CXR confirmed the clinical diagnosis and allowed an assessment of size to guide treatment. Routine CXR after permanent pacemaker insertion is not necessary in uncomplicated cases with adequate pacing characteristics.


Evaluation of surface characteristics of rotary nickel-titanium instruments produced by different manufacturing methods.
Instrument fracture is a serious concern in endodontic practice. The aim of this study was to investigate the surface quality of new and used rotary nickel-titanium (NiTi) instruments manufactured by the traditional grinding process and twisting methods. Total 16 instruments of two rotary NiTi systems were used in this study. Eight Twisted Files (TF) (SybronEndo, Orange, CA, USA) and 8 Mtwo (VDW, Munich, Germany) instruments were evaluated. New and used of 4 experimental groups were evaluated using an atomic force microscopy (AFM). New and used instruments were analyzed on 3 points along a 3 mm. section at the tip of the instrument. Quantitative measurements according to the topographical deviations were recorded. The data were statistically analyzed with paired samples t-test and independent samples t-test. Mean root mean square (RMS) values for new and used TF 25.06 files were 10.70 ± 2.80 nm and 21.58 ± 6.42 nm, respectively, and the difference between them was statistically significant (P < 0.05). Mean RMS values for new and used Mtwo 25.06 files were 24.16 ± 9.30 nm and 39.15 ± 16.20 nm respectively, the difference between them also was statistically significant (P < 0.05). According to the AFM analysis, instruments produced by twisting method (TF 25.06) had better surface quality than the instruments produced by traditional grinding process (Mtwo 25.06 files).


The southern route "out of Africa": evidence for an early expansion of modern humans into Arabia.
The timing of the dispersal of anatomically modern humans (AMH) out of Africa is a fundamental question in human evolutionary studies. Existing data suggest a rapid coastal exodus via the Indian Ocean rim around 60,000 years ago. We present evidence from Jebel Faya, United Arab Emirates, demonstrating human presence in eastern Arabia during the last interglacial. The tool kit found at Jebel Faya has affinities to the late Middle Stone Age in northeast Africa, indicating that technological innovation was not necessary to facilitate migration into Arabia. Instead, we propose that low eustatic sea level and increased rainfall during the transition between marine isotope stages 6 and 5 allowed humans to populate Arabia. This evidence implies that AMH may have been present in South Asia before the Toba eruption.


Impact of Oral Anticoagulation Therapy Versus Left Atrial Appendage Occlusion on Cognitive Function and Quality of Life in Patients With Atrial Fibrillation.
Background We compared the cognitive status and quality of life in patients with atrial fibrillation undergoing left atrial appendage occlusion (LAAO) or remaining on oral anticoagulation (OAC) after atrial fibrillation ablation. Methods and Results Cognition was assessed by the Montreal Cognitive Assessment (MoCA) survey at baseline and follow-up. Consecutive patients receiving LAAO or OAC after atrial fibrillation ablation were screened, and patients with a score of ≤17 were excluded from the study. Quality of life was measured at baseline and 1 year using the Atrial Fibrillation Effect on Quality of Life survey. A total of 50 patients (CHA2DS2-VASc [congestive heart failure, hypertension, age≥75 years, diabetes mellitus, stroke or transient ischemic attack, vascular disease, age 65-74 years, sex category] score: 3.30±1.43) in the LAAO group and 48 (CHA2DS2-VASc score 2.73±1.25) in the OAC group were included in this prospective study. Mean baseline MoCA score was 26.18 and 26.08 in the LAAO and OAC groups, respectively (P=0.846). At 1 year, scores were 26.94 and 23.38 in the respective groups. MoCA score decreased by an estimated -2.74 (95% CI, -3.61 to -1.87; P<0.0001) points in the OAC group, whereas the change in the LAAO group was nonsignificant (0.79; (95% CI, -0.06 to 1.64; P=0.07). After adjusting for baseline clinical characteristics, remaining on OAC was an independent predictor of MoCA change at 1 year (regression coefficient, -3.38; 95% CI, -4.75 to -2.02; P<0.0001). Change in Atrial Fibrillation Effect on Quality of Life score did not differ significantly in achieving a clinically important difference between groups. Conclusions In this series, a significant difference in the postprocedure MoCA score was observed in postablation patients with atrial fibrillation receiving LAAO versus remaining on OAC with a substantial decline in the score in the OAC group. However, quality of life improved similarly across groups. Registration https://www.ClinicalTrials.gov. Unique identifier: NCT01816308.


Prospective in-patient cohort study of moves between levels of therapeutic security: the DUNDRUM-1 triage security, DUNDRUM-3 programme completion and DUNDRUM-4 recovery scales and the HCR-20.
We examined whether new structured professional judgment instruments for assessing need for therapeutic security, treatment completion and recovery in forensic settings were related to moves from higher to lower levels of therapeutic security and added anything to assessment of risk. This was a prospective naturalistic twelve month observational study of a cohort of patients in a forensic hospital placed according to their need for therapeutic security along a pathway of moves from high to progressively less secure units in preparation for discharge. Patients were assessed using the DUNDRUM-1 triage security scale, the DUNDRUM-3 programme completion scale and the DUNDRUM-4 recovery scale and assessments of risk of violence, self harm and suicide, symptom severity and global function. Patients were subsequently observed for positive moves to less secure units and negative moves to more secure units. There were 86 male patients at baseline with mean follow-up 0.9 years, 11 positive and 9 negative moves. For positive moves, logistic regression indicated that along with location at baseline, the DUNDRUM-1, HCR-20 dynamic and PANSS general symptom scores were associated with subsequent positive moves. The receiver operating characteristic was significant for the DUNDRUM-1 while ANOVA co-varying for both location at baseline and HCR-20 dynamic score was significant for DUNDRUM-1. For negative moves, logistic regression showed DUNDRUM-1 and HCR-20 dynamic scores were associated with subsequent negative moves, along with DUNDRUM-3 and PANSS negative symptoms in some models. The receiver operating characteristic was significant for the DUNDRUM-4 recovery and HCR-20 dynamic scores with DUNDRUM-1, DUNDRUM-3, PANSS general and GAF marginal. ANOVA co-varying for both location at baseline and HCR-20 dynamic scores showed only DUNDRUM-1 and PANSS negative symptoms associated with subsequent negative moves. Clinicians appear to decide moves based on combinations of current and imminent (dynamic) risk measured by HCR-20 dynamic score and historical seriousness of risk as measured by need for therapeutic security (DUNDRUM-1) in keeping with Scott's formulation of risk and seriousness. The DUNDRUM-3 programme completion and DUNDRUM-4 recovery scales have utility as dynamic measures that can off-set perceived 'dangerousness'.


Effects of dog-assisted intervention on behavioural and psychological symptoms of dementia.
To evaluate the effect of a dog-assisted intervention on the behavioural and psychological symptoms of residents with dementia during a six-month period. The study was conducted in eight nursing homes in Sweden. A total of 33 residents with dementia, 20 in the intervention group and 13 in the control group, were recruited. The Cohen-Mansfield Agitation Inventory (CMAI) and the Multi-Dimensional Dementia Assessment Scale (MDDAS) were used to assess the effects of a dog-assisted intervention on participants' behavioural and psychological symptoms. The intervention comprised ten sessions, lasting between 45 and 60 minutes, once or twice a week. Descriptive statistics were used to analyse background data, comparisons between groups at baseline were performed using the Mann-Whitney U test, and the Wilcoxon rank sum test was used to test differences in groups over time. In the intervention group changes from baseline to follow up immediately after the intervention were not significant, possibly because of the small sample size. Some positive tendencies were observed: the CMAI mean score for physical non-aggressive behaviours decreased from 18.5 at baseline to 15.3 at follow up immediately after the intervention; lower scores indicate fewer symptoms. Mean and median MDDAS scores for behavioural symptoms decreased from 15.3 and 13.5 respectively at baseline to 13.1 and 12.0 respectively at follow up immediately after the intervention; lower scores indicate fewer symptoms. The CMAI mean score for verbal agitation increased significantly (P=0.035) from 17.2 at baseline to 20.6 at follow up six months after the intervention. Dog-assisted intervention may provide an alternative or a complement to pharmacological treatments to reduce behavioural symptoms in people with dementia, but its value and place in care require further evaluation.


IGF1R and Src inhibition induce synergistic cytotoxicity in HNSCC through inhibition of FAK.
Head and neck cancer is the sixth most common cancer worldwide with a 5-year survival of only 65%. Targeting compensatory signaling pathways may improve therapeutic responses and combat resistance. Utilizing reverse phase protein arrays (RPPA) to assess the proteome and explore mechanisms of synergistic growth inhibition in HNSCC cell lines treated with IGF1R and Src inhibitors, BMS754807 and dasatinib, respectively, we identified focal adhesion signaling as a critical node. Focal Adhesion Kinase (FAK) and Paxillin phosphorylation were decreased as early as 15 min after treatment, and treatment with a FAK inhibitor, PF-562,271, was sufficient to decrease viability in vitro. Treatment of 3D spheroids demonstrated robust cytotoxicity suggesting that the combination of BMS754807 and dasatinib is effective in multiple experimental models. Furthermore, treatment with BMS754807 and dasatinib significantly decreased cell motility, migration, and invasion in multiple HNSCC cell lines. Most strikingly, treatment with BMS754807 and dasatinib, or a FAK inhibitor alone, significantly increased cleaved-PARP in human ex-vivo HNSCC patient tissues demonstrating a potential clinical utility for targeting FAK or the combined targeting of the IGF1R with Src. This ex-vivo result further confirms FAK as a vital signaling node of this combinatorial treatment and demonstrates therapeutic potential for targeting FAK in HNSCC patients.


Travertine crystal growth ripples record the hydraulic history of ancient Rome's Anio Novus aqueduct.
Travertine crystal growth ripples are used to reconstruct the early hydraulic history of the Anio Novus aqueduct of ancient Rome. These crystalline morphologies deposited within the aqueduct channel record the hydraulic history of gravity-driven turbulent flow at the time of Roman operation. The wavelength, amplitude, and steepness of these travertine crystal growth ripples indicate that large-scale sustained aqueduct flows scaled directly with the thickness of the aqueous viscous sublayer. Resulting critical shear Reynolds numbers are comparable with those reconstructed from heat/mass transfer crystalline ripples formed in other natural and engineered environments. This includes sediment transport in rivers, lakes, and oceans, chemical precipitation and dissolution in caves, and melting and freezing in ice. Where flow depth and perimeter could be reconstructed from the distribution and stratigraphy of the travertine within the Anio Novus aqueduct, flow velocity and rate have been quantified by deriving roughness-flow relationships that are independent of water temperature. More generally, under conditions of near-constant water temperature and kinematic viscosity within the Anio Novus aqueduct channel, the travertine crystal growth ripple wavelengths increased with decreasing flow velocity, indicating that systematic changes took place in flow rate during travertine deposition. This study establishes that travertine crystal growth ripples such as those preserved in the Anio Novus provide a sensitive record of past hydraulic conditions, which can be similarly reconstructed from travertine deposited in other ancient water conveyance and storage systems around the world.


Do fundamental fears differentially contribute to pain-related fear and pain catastrophizing? An evaluation of the sensitivity index.
Three fundamental fears - anxiety sensitivity (AS), injury/illness sensitivity (IS) and fear of negative evaluation (FNE) - have been proposed to underlie common fears and psychopathological conditions. In pain research, the relation between AS and (chronic) pain processes was the subject of several studies, whereas the possible role of IS has been ignored. The current research examines the role of IS with respect to various pain-related variables in two studies. In the first study, 192 healthy college students completed the Sensitivity Index (SI; a composite measure assessing the three fundamental fears) and various pain-related questionnaires. In a second study, 60 students out of the original sample took part in a pain induction procedure and completed the SI as well. We first examined the properties of the SI. Factor analysis on the SI replicated the proposed factor structure [Taylor S. The structure of fundamental fears, J Behav Ther Exp Psychiat 1993;24:289-99]. However, some items of the ASI did show problematic loadings and were therefore excluded in subsequent analyses. The main hypothesis of the current study states that IS is a stronger predictor than AS of pain catastrophizing and fear of pain as assessed by self-report measures, and of pain tolerance and anticipatory fear of pain as assessed in a pain induction study. This hypothesis could be confirmed for all variables, except for pain tolerance, which was not predicted by any of the three fundamental fears. The current study can be considered as an impetus for devoting attention to IS in future pain research.


Catechol-O-methyltransferase inhibition alters pain and anxiety-related volitional behaviors through activation of β-adrenergic receptors in the rat.
Reduced catechol-O-methyltransferase (COMT) activity resulting from genetic variation or pharmacological depletion results in enhanced pain perception in humans and nociceptive behaviors in animals. Using phasic mechanical and thermal reflex tests (e.g. von Frey, Hargreaves), recent studies show that acute COMT-dependent pain in rats is mediated by β-adrenergic receptors (βARs). In order to more closely mimic the characteristics of human chronic pain conditions associated with prolonged reductions in COMT, the present study sought to determine volitional pain-related and anxiety-like behavioral responses following sustained as well as acute COMT inhibition using an operant 10-45°C thermal place preference task and a light/dark preference test. In addition, we sought to evaluate the effects of sustained COMT inhibition on generalized body pain by measuring tactile sensory thresholds of the abdominal region. Results demonstrated that acute and sustained administration of the COMT inhibitor OR486 increased pain behavior in response to thermal heat. Further, sustained administration of OR486 increased anxiety behavior in response to bright light, as well as abdominal mechanosensation. Finally, all pain-related behaviors were blocked by the non-selective βAR antagonist propranolol. Collectively, these findings provide the first evidence that stimulation of βARs following acute or chronic COMT inhibition drives cognitive-affective behaviors associated with heightened pain that affects multiple body sites.


Guidance for the treatment of deep vein thrombosis and pulmonary embolism.
This guidance document focuses on the diagnosis and treatment of venous thromboembolism (VTE). Efficient, cost effective diagnosis of VTE is facilitated by combining medical history and physical examination with pre-test probability models, D dimer testing and selective use of confirmatory imaging. Clinical prediction rules, biomarkers and imaging can be used to tailor therapy to disease severity. Anticoagulation options for acute VTE include unfractionated heparin, low molecular weight heparin, fondaparinux and the direct oral anticoagulants (DOACs). DOACs are as effective as conventional therapy with LMWH and vitamin K antagonists. Thrombolytic therapy is reserved for massive pulmonary embolism (PE) or extensive deep vein thrombosis (DVT). Inferior vena cava filters are reserved for patients with acute VTE and contraindications to anticoagulation. Retrievable filters are strongly preferred. The possibility of thoracic outlet syndrome and May-Thurner syndrome should be considered in patients with subclavian/axillary and left common iliac vein DVT, respectively in absence of identifiable triggers. The optimal duration of therapy is dictated by the presence of modifiable thrombotic risk factors. Long term anticoagulation should be considered in patients with unprovoked VTE as well as persistent prothrombotic risk factors such as cancer. Short-term therapy is sufficient for most patients with VTE associated with transient situational triggers such as major surgery. Biomarkers such as D dimer and risk assessment models such the Vienna risk prediction model offer the potential to customize VTE therapy for the individual patient. Insufficient data exist to support the integration of bleeding risk models into duration of therapy planning.


A closer look at CD1d molecules: new horizons in studying NKT cells.
Recent findings have highlighted the ability of invariant natural killer T (iNKT) cells to recognize microbe-derived glycolipids and have demonstrated the role of these cells in several disease states, from autoimmune disease to cancer. It has also become clear that iNKT cells can rapidly mature dendritic cells and licence them to prime antigen-specific T- and B-cell responses. The use of CD1d tetramers to monitor iNKT cell frequency and phenotype has moved the field forward at a fast pace. To harness iNKT cells for therapeutic purposes and to understand their role in vivo, it is essential to characterize the molecular events that contribute to iNKT cell activation. Here we review new reagents and novel protocols that are facilitating a closer look at lipid presentation by CD1d molecules and their recognition by iNKT cells.


National outcomes of in-hospital cardiopulmonary resuscitation in patients with acute kidney injury: years 2005 - 2011 .
There is paucity of data on the outcomes of in-hospital cardiopulmonary resuscitation (CPR) in patients with acute kidney injury (AKI). We analyzed the impact of acute kidney injury on in-hospital CPR-related outcomes. We analyzed data from Nationwide Inpatient Sample (NIS 2005 - 2011) including patients with and without AKI who had undergone in-hospital CPR. Baseline characteristics, in-hospital complications and discharge outcomes were compared between the two groups. We determined the effect of AKI on length of hospital stay, discharge destination, hospital mortality, survival trends, and discharge to home. 180,970 patients with primary or secondary diagnosis of AKI underwent in-hospital CPR compared to 323,620 patients without AKI. Unadjusted in-hospital mortality rates were higher in the AKI group (78.2 vs. 71.8%, p < 0.0001). After adjusting for age, sex, and potential confounders, patients in the AKI group had higher odds of mortality with odds ratio 1.3, 95% confidence interval 1.2 - 1.4, p < 0.0001. Survivors in the AKI group were more likely to be discharged to nursing homes and had higher mean hospitalization charges. In 2011 compared with 2005, there was an improved survival after CPR and higher rates of discharges to home. There was no significant change in the mean length of hospital stay between these time periods (p = 0.4). AKI independently increases the odds of in-hospital mortality and nursing home placement after in-hospital CPR. These data may facilitate CPR discussions and decision-making in critically ill patients. .


Risk factors for wound-related reoperations in patients with metastatic spine tumor.
OBJECTIVE Resection of metastatic spine tumors can improve patients' quality of life by addressing pain or neurological compromise. However, resections are often complicated by wound dehiscence, infection, instrumentation failures, and the need for reoperation. Moreover, when reoperations are needed, the most common indication is surgical site infection and wound breakdown. In turn, wound reoperations increase morbidity as well as the length and cost of hospitalization. The aim of this study was to examine perioperative risk factors associated with increased rate of wound reoperations after metastatic spine tumor resection. METHODS A retrospective study of patients at a single institution who underwent metastatic spine tumor resection between 2003 and 2013 was conducted. Factors with a p value < 0.200 in a univariate analysis were included in the multivariate model. RESULTS A total of 159 patients were included in this study. Karnofsky Performance Scale score > 70, smoking status, hypertension, thromboembolic events, hyperlipidemia, increasing number of vertebral levels, and posterior approach were included in the multivariate analysis. Thromboembolic events (95% CI 1.19-48.5, p = 0.032) and number of levels involved were independently associated with increased wound reoperation rates in the multivariate model. For each additional spinal level involved, the risk for wound reoperations increased by 21% (95% CI 1.03-1.43, p = 0.018). CONCLUSIONS Although wound complications and subsequent reoperations are potential risks for all patients with metastatic spine tumor, due to adjuvant radiotherapy and other medical comorbidities, this study identified patients with thromboembolic events or those requiring a larger incision as being at the highest risk. Measures intended to decrease the occurrence of perioperative venous thromboembolism and to improve wound care, especially for long incisions, may decrease wound-related revision surgeries in this vulnerable group of patients.


Inclusion of non-viable neonates in the birth record and its impact on infant mortality rates in Shelby County, Tennessee, USA.
Rates of infant death are one of the most common indicators of a population's overall health status. Infant mortality rates (IMRs) are used to make broad inferences about the quality of health care, effects of health policies and even environmental quality. The purpose of our study was threefold: i) to examine the characteristics of births in the area in relation to gestational age and birthweight; ii) to estimate infant mortality using variable gestational age and/or birthweight criteria for live birth, and iii) to calculate proportional mortality ratios for each cause of death using variable gestational age and/or birthweight criteria for live birth. We conducted a retrospective analysis of all Shelby County resident-linked birth and infant death certificates during the years 1999 to 2004. Descriptive test statistics were used to examine infant mortality rates in relation to specific maternal and infant risk factors. Through careful examination of 1999-2004 resident-linked birth and infant death data sets, we observed a disproportionate number of non-viable live births (≤20 weeks gestation or ≤350 grams) in Shelby County. Issuance of birth certificates to these non-viable neonates is a factor that contributes to an inflated IMR. Our study demonstrates the complexity and the appropriateness of comparing infant mortality rates in smaller geographic units, given the unique characteristics of live births in Shelby County. The disproportionate number of pre-viable infants born in Shelby County greatly obfuscates neonatal mortality and de-emphasizes the importance of post-neonatal mortality.


Vitamin A transport and the transmembrane pore in the cell-surface receptor for plasma retinol binding protein.
Vitamin A and its derivatives (retinoids) play diverse and crucial functions from embryogenesis to adulthood and are used as therapeutic agents in human medicine for eye and skin diseases, infections and cancer. Plasma retinol binding protein (RBP) is the principal and specific vitamin A carrier in the blood and binds vitamin A at 1:1 ratio. STRA6 is the high-affinity membrane receptor for RBP and mediates cellular vitamin A uptake. STRA6 null mice have severely depleted vitamin A reserves for vision and consequently have vision loss, even under vitamin A sufficient conditions. STRA6 null humans have a wide range of severe pathological phenotypes in many organs including the eye, brain, heart and lung. Known membrane transport mechanisms involve transmembrane pores that regulate the transport of the substrate (e.g., the gating of ion channels). STRA6 represents a new type of membrane receptor. How this receptor interacts with its transport substrate vitamin A and the functions of its nine transmembrane domains are still completely unknown. These questions are critical to understanding the molecular basis of STRA6's activities and its regulation. We employ acute chemical modification to introduce chemical side chains to STRA6 in a site-specific manner. We found that modifications with specific chemicals at specific positions in or near the transmembrane domains of this receptor can almost completely suppress its vitamin A transport activity. These experiments provide the first evidence for the existence of a transmembrane pore, analogous to the pore of ion channels, for this new type of cell-surface receptor.


The HMG-domain protein BAP111 is important for the function of the BRM chromatin-remodeling complex in vivo.
The Drosophila trithorax group gene brahma (brm) encodes the ATPase subunit of a SWI/SNF-like chromatin-remodeling complex. A key question about chromatin-remodeling complexes is how they interact with DNA, particularly in the large genomes of higher eukaryotes. Here, we report the characterization of BAP111, a BRM-associated protein that contains a high mobility group (HMG) domain predicted to bind distorted or bent DNA. The presence of an HMG domain in BAP111 suggests that it may modulate interactions between the BRM complex and chromatin. BAP111 is an abundant nuclear protein that is present in all cells throughout development. By using gel filtration chromatography and immunoprecipitation assays, we found that the majority of BAP111 protein in embryos is associated with the BRM complex. Furthermore, heterozygosity for BAP111 enhanced the phenotypes resulting from a partial loss of brm function. These data demonstrate that the BAP111 subunit is important for BRM complex function in vivo.


Simian sarcoma virus--transformed cells secrete a mitogen identical to platelet-derived growth factor.
Normal rat kidney (NRK) cells transformed by simian sarcoma virus (SSV) release into the culture medium a biologically active mitogen with properties identical to those of human platelet-derived growth factor (PDGF). Like PDGF, the growth factor derived from SSV-NRK cells was shown to be stable to heat and sensitive to reducing agents. It was capable of inhibiting binding of labeled PDGF to the receptor on human fibroblasts. It also stimulated the phosphorylation of the same membrane protein (185 kilodaltons) in isolated plasma membranes from human fibroblasts. Immunoprecipitation of metabolically labeled proteins released by SSV-NRK cells showed that a 34-kilodalton protein was specifically precipitated by antiserum to PDGF. Upon reduction, this protein had a molecular size of 17 kilodaltons. PDGF has been shown to consist of two 14- to 18-kilodalton proteins linked by disulfide bonds.


Targeting p21-activated kinase 1 for development of a novel anti-arrhythmic drug class.
Evidence accumulated over the past decade suggests that p21-activated kinase 1 (PAK1) is a critical cardiac-protective signalling molecule. The present article provides an updated review of recent findings regarding the role of PAK1 in maintaining normal cardiac electrophysiological function through its regulation of membrane and Ca2+ clocks. We first overviewed the PAK1 activation mechanism. We then discussed recent updated results showing the action mechanisms of PAK1 signalling on Cav1.2/Cav1.3 (ICaL)-mediated Ca2+ entry, ryanodine receptor type 2-mediated sarcoplasmic reticulum (SR) Ca2+ release, transcriptional regulation of SR Ca2+-ATPase 2a, Na+/Ca2+ exchangers, and Ca2+/calmodulin-dependent protein kinase II. Finally, we proposed a new and exciting route for developing a PAK1-based therapeutic strategy for cardiac arrhythmias. This article is part of the theme issue 'The heartbeat: its molecular basis and physiological mechanisms'.


Does the frequency of respiratory tract infections help to identify humoral immunodeficiencies in a primary health-care cohort?
Primary immune deficiency (PID) due to humoral defects is associated with recurrent respiratory tract infections (RTIs). Reliable clinical warning signs of PID would facilitate early diagnosis and thereby reduce long-term complications. The aim of the present study was to evaluate the accuracy of the warning sign, 'four or more antibiotic-treated RTIs annually for 3 or more consecutive years,' for detecting PID among adults in a primary health-care setting. Fifty-three cases with 'four or more antibiotic-treated RTIs annually for 3 or more consecutive years' were selected from a Swedish primary health-care registry of RTIs. In addition, 66 age- and sex-matched controls were selected having a maximum of one antibiotic-treated RTI during the period covered by the study. Levels of immunoglobulin (Ig) IgG, IgA, IgM, IgG subclasses, and IgG antibodies against Haemophilus influenzae and Streptococcus pneumoniae as well as the inflammatory markers, C-reactive protein, interleukin (IL)-6 and IL-8 were determined. IgG subclass deficiencies (IgGsd) were found in 5/53 (9.4%) of the cases and in 7/66 (10.6%) controls. The most frequent deficiency was IgG3sd and this was found in three participants in the case group and seven in the control group. The mean level of IgG3 was lower in the control group (p = 0.02). The mean level of IL-8 was lower in the case group (p = 0.02). The results show that physicians working in primary health care cannot solely rely on the frequency of antibiotic-treated RTIs as a warning sign for the detection of common humoral immune deficiencies.


Roles of key active-site residues in flavocytochrome P450 BM3.
The effects of mutation of key active-site residues (Arg-47, Tyr-51, Phe-42 and Phe-87) in Bacillus megaterium flavocytochrome P450 BM3 were investigated. Kinetic studies on the oxidation of laurate and arachidonate showed that the side chain of Arg-47 contributes more significantly to stabilization of the fatty acid carboxylate than does that of Tyr-51 (kinetic parameters for oxidation of laurate: R47A mutant, Km 859 microM, kcat 3960 min-1; Y51F mutant, Km 432 microM, kcat 6140 min-1; wild-type, Km 288 microM, kcat 5140 min-1). A slightly increased kcat for the Y51F-catalysed oxidation of laurate is probably due to decreased activation energy (DeltaG) resulting from a smaller DeltaG of substrate binding. The side chain of Phe-42 acts as a phenyl 'cap' over the mouth of the substrate-binding channel. With mutant F42A, Km is massively increased and kcat is decreased for oxidation of both laurate (Km 2. 08 mM, kcat 2450 min-1) and arachidonate (Km 34.9 microM, kcat 14620 min-1; compared with values of 4.7 microM and 17100 min-1 respectively for wild-type). Amino acid Phe-87 is critical for efficient catalysis. Mutants F87G and F87Y not only exhibit increased Km and decreased kcat values for fatty acid oxidation, but also undergo an irreversible conversion process from a 'fast' to a 'slow' rate of substrate turnover [for F87G (F87Y)-catalysed laurate oxidation: kcat 'fast', 760 (1620) min-1; kcat 'slow', 48.0 (44.6) min-1; kconv (rate of conversion from fast to slow form), 4.9 (23.8) min-1]. All mutants showed less than 10% uncoupling of NADPH oxidation from fatty acid oxidation. The rate of FMN-to-haem electron transfer was shown to become rate-limiting in all mutants analysed. For wild-type P450 BM3, the rate of FMN-to-haem electron transfer (8340 min-1) is twice the steady-state rate of oxidation (4100 min-1), indicating that other steps contribute to rate limitation. Active-site structures of the mutants were probed with the inhibitors 12-(imidazolyl)dodecanoic acid and 1-phenylimidazole. Mutant F87G binds 1-phenylimidazole >10-fold more tightly than does the wild-type, whereas mutant Y51F binds the haem-co-ordinating fatty acid analogue 12-(imidazolyl)dodecanoic acid >30-fold more tightly than wild-type.


Understanding the production of population health and the role of paying for population health.
This article considers 2 related themes that address population health outcomes and the contributions to those outcomes by time, place, individual behaviors and choices, and activities of various social sectors. First, what does it mean to "produce" population health, and how can the production of health be understood empirically? Second, through what processes can incentives be modified to improve population health? Among the issues that arise are understanding the mechanisms through which paying for population health works and how the health-producing incentives materialize in various sectors, especially those whose primary functions are not generally viewed as fostering better population health.


Semantic relevance and semantic disorders.
Semantic features are of different importance in concept representation. The concept elephant may be more easily identified from the feature <trunk> than from the feature <four legs>. We propose a new model of semantic memory to measure the relevance of semantic features for a concept and use this model to investigate the controversial issue of category specificity. Category-specific patients have an impairment in one domain of knowledge (e.g., living), whereas the other domain (e.g., nonliving) is relatively spared. We show that categories differ in the level of relevance and that, when concepts belonging to living and nonliving categories are equated to this parameter, the category-specific disorder disappears. Our findings suggest that category specificity, as well as other semantic-related effects, may be explained by a semantic memory model in which concepts are represented by semantic features with associated relevance values.


Genome-wide screening of potential RNase Y-processed mRNAs in the M49 serotype Streptococcus pyogenes NZ131.
RNase Y is a major endoribonuclease in Group A streptococcus (GAS) and other Gram-positive bacteria. Our previous study showed that RNase Y was involved in mRNA degradation and processing in GAS. We hypothesized that mRNA processing regulated the expression of important GAS virulence factors via altering their mRNA stabilities and that RNase Y mediated at least some of the mRNA-processing events. The aims of this study were to (1) identify mRNAs that were processed by RNase Y and (2) confirm the mRNA-processing events. The transcriptomes of Streptococcus pyogenes NZ131 wild type and its RNase Y mutant (Δrny) were examined with RNA-seq. The data were further analyzed to define GAS operons. The mRNA stabilities of the wild type and Δrny at subgene level were determined with tiling array analysis. Operons displaying segmental stability in the wild type but not in the Δrny were predicted to be RNase Y processed. Overall 865 operons were defined and their boundaries predicted. Further analysis narrowed down 15 mRNAs potentially processed by RNase Y. A selection of four candidates including folC1 (folylpolyglutamate synthetase), prtF (fibronectin-binding protein), speG (streptococcal exotoxin G), ropB (transcriptional regulator of speB), and ypaA (riboflavin transporter) mRNAs was examined with Northern blot analysis. However, only folC1 was confirmed to be processed, but it is unlikely that RNase Y is responsible. We conclude that GAS use RNase Y to selectively process mRNA, but the overall impact is confined to selected virulence factors.


Modification of seed oil content and acyl composition in the brassicaceae by expression of a yeast sn-2 acyltransferase gene.
A putative yeast sn-2 acyltransferase gene (SLC1-1), reportedly a variant acyltransferase that suppresses a genetic defect in sphingolipid long-chain base biosynthesis, has been expressed in a yeast SLC deletion strain. The SLC1-1 gene product was shown in vitro to encode an sn-2 acyltransferase capable of acylating sn-1 oleoyl-lysophosphatidic acid, using a range of acyl-CoA thioesters, including 18:1-, 22:1-, and 24:0-CoAs. The SLC1-1 gene was introduced into Arabidopsis and a high erucic acid-containing Brassica napus cv Hero under the control of a constitutive (tandem cauliflower mosaic virus 35S) promoter. The resulting transgenic plants showed substantial increases of 8 to 48% in seed oil content (expressed on the basis of seed dry weight) and increases in both overall proportions and amounts of very-long-chain fatty acids in seed triacylglycerols (TAGs). Furthermore, the proportion of very-long-chain fatty acids found at the sn-2 position of TAGs was increased, and homogenates prepared from developing seeds of transformed plants exhibited elevated lysophosphatidic acid acyltransferase (EC 22.214.171.124) activity. Thus, the yeast sn-2 acyltransferase has been shown to encode a protein that can exhibit lysophosphatidic acid acyltransferase activity and that can be used to change total fatty acid content and composition as well as to alter the stereospecific acyl distribution of fatty acids in seed TAGs.


Suppression of in vitro cytotoxic response by macrophages due to induced arginase.
Arginine was found to be completely depleted from cell-free supernates of mixed leukocyte cultures suppressed by the addition of excess macrophages. Partial reversal of macrophage-mediated suppression was accomplished by daily addition of a cocktail containing arginine and nonamino acid nutrients. Complete reversal of the suppression was accomplished by the addition of arginine and glucose to the medium and the nonadherent cells after their separation from the adherent macrophages. A marked increase in the enzyme arginase was found in macrophages that had been cultured 24 h in vitro in Eagle's minimum essential medium plus 10% fetal calf serum, in peritoneal cells activated by prior injection of thioglycollate, and in one spleen activated by a graft vs. host reaction.


Perceptual Learning of Vocoded Speech With and Without Contralateral Hearing: Implications for Cochlear Implant Rehabilitation.
Purpose An increasing number of individuals with residual or even normal contralateral hearing are being considered for cochlear implantation. It remains unknown whether the presence of contralateral hearing is beneficial or detrimental to their perceptual learning of cochlear implant (CI)-processed speech. The aim of this experiment was to provide a first insight into this question using acoustic simulations of CI processing. Method Sixty normal-hearing listeners took part in an auditory perceptual learning experiment. Each subject was randomly assigned to one of three groups of 20 referred to as NORMAL, LOWPASS, and NOTHING. The experiment consisted of two test phases separated by a training phase. In the test phases, all subjects were tested on recognition of monosyllabic words passed through a six-channel "PSHC" vocoder presented to a single ear. In the training phase, which consisted of listening to a 25-min audio book, all subjects were also presented with the same vocoded speech in one ear but the signal they received in their other ear differed across groups. The NORMAL group was presented with the unprocessed speech signal, the LOWPASS group with a low-pass filtered version of the speech signal, and the NOTHING group with no sound at all. Results The improvement in speech scores following training was significantly smaller for the NORMAL than for the LOWPASS and NOTHING groups. Conclusions This study suggests that the presentation of normal speech in the contralateral ear reduces or slows down perceptual learning of vocoded speech but that an unintelligible low-pass filtered contralateral signal does not have this effect. Potential implications for the rehabilitation of CI patients with partial or full contralateral hearing are discussed.


Mucosal barrier injury laboratory-confirmed bloodstream infection: results from a field test of a new National Healthcare Safety Network definition.
To assess challenges to implementation of a new National Healthcare Safety Network (NHSN) surveillance definition, mucosal barrier injury laboratory-confirmed bloodstream infection (MBI-LCBI). Multicenter field test. Selected locations of acute care hospitals participating in NHSN central line-associated bloodstream infection (CLABSI) surveillance. Hospital staff augmented their CLABSI surveillance for 2 months to incorporate MBI-LCBI: a primary bloodstream infection due to a selected group of organisms in patients with either neutropenia or an allogeneic hematopoietic stem cell transplant with gastrointestinal graft-versus-host disease or diarrhea. Centers for Disease Control and Prevention (CDC) staff reviewed submitted data to verify whether CLABSIs met MBI-LCBI criteria and summarized the descriptive epidemiology of cases reported. Eight cancer, 2 pediatric, and 28 general acute care hospitals including 193 inpatient units (49% oncology/bone marrow transplant [BMT], 21% adult ward, 20% adult critical care, 6% pediatric, 4% step-down) conducted field testing. Among 906 positive blood cultures reviewed, 282 CLABSIs were identified. Of the 103 CLABSIs that also met MBI-LCBI criteria, 100 (97%) were reported from oncology/BMT locations. Agreement between hospital staff and CDC classification of reported CLABSIs as meeting the MBI-LCBI definition was high (90%; κ = 0.82). Most MBI-LCBIs (91%) occurred in patients meeting neutropenia criteria. Some hospitals indicated that their laboratories' methods of reporting cell counts prevented application of neutropenia criteria; revised neutropenia criteria were created using data from field testing. Hospital staff applied the MBI-LCBI definition accurately. Field testing informed modifications for the January 2013 implementation of MBI-LCBI in the NHSN.


Effects of food marketing to kids: I'm lovin' it?
This issue brief reviews key findings and recommendations from the Institute of Medicine study on food marketing and its effects on childhood obesity. The brief describes the childhood obesity epidemic, discusses key trends associated with rising childhood obesity rates, and considers the relative role of marketing practices on diet and obesity within the broader context of complex contributory factors. The brief also summarizes the current legal framework for regulating marketing directed at children;discusses voluntary, self-regulatory mechanisms; and highlights proposals to re-orient marketing practices to combat childhood obesity.


No need to change the skin knife in modern arthroplasty surgery.
Earlier studies have found varying contamination rates using separate skin and deep knives in total hip (THA) and total knee (TKA) arthroplasty surgery. Previous studies were primarily conducted in the setting of concomitant use of laminar airflow and/or plastic adhesive draping. This has lead to conflicting conclusions regarding discarding the skin knife or not. This study evaluates the prevalence of contamination of a separate skin knife using modern antiseptic technique in primary THA and TKA without laminar airflow. Three knives from each primary THA and TKA surgery in non-laminar airflow operating rooms were collected: one used for the skin, one used for deeper tissues and one control knife. A total of 831 knife blades from 277 patients were cultured 12 days. Contamination of the skin knife was found in eight patients (2.8 %), contamination of the "deep" knife in five patients (1.8 %) and contamination of the control knife in five patients (1.8 %). No patient developed an infection with 1-year follow-up. Our findings suggest a very low rate of contamination of the skin knife using modern antiseptic technique without laminar airflow and/or plastic adhesive draping and do not support the use of a separate skin knife in arthroplasty surgery.


Genetic tools for studying adaptation and the evolution of behavior.
The rapid expansion of genomic and molecular genetic techniques in model organisms, and the application of these techniques to organisms that are less well studied genetically, make it possible to understand the genetic control of many behavioral phenotypes. However, many behavioral ecologists are uncertain about the value of including a genetic component in their studies. In this article, we review how genetic analyses of behavior are central to topics ranging from understanding past selection and predicting future evolution to explaining the neural and hormonal control of behavior. Furthermore, we review both new and old techniques for studying evolutionary behavior genetics and highlight how the choice of approach depends on both the question and the organism. Topics discussed include genetic architecture, detecting the past history of selection, and genotype-by-environment interactions. We show how these questions are being addressed with techniques including statistical genetics, QTL analyses, transgenic analyses, and microarrays. Many of the techniques were first applied to the behavior of genetic model organisms such as laboratory mice and flies. Two recent developments serve to expand the relevance of such studies to behavioral ecology. The first is to use model organisms for studies of the genetic basis of evolutionarily relevant behavior and the second is to apply methods developed in model genetic systems to species that have not previously been examined genetically. These conceptual advances, along with the rapid diversification of genetic tools and the recognition of widespread genetic homology, suggest a bright outlook for evolutionary genetic studies. This review provides access to tools through references to the recent literature and shows the great promise for evolutionary behavioral genetics.


Learning enhances the survival of new neurons beyond the time when the hippocampus is required for memory.
Trace memories are formed when a stimulus event becomes associated with another event that occurs later in time and is discontinuous with the first event. The formation of trace memories enhances the survival of newly generated neurons in the dentate gyrus of the adult hippocampus (Gould et al., 1999a). Here we tested whether the acquisition of trace memories early during training is sufficient to enhance cell survival. We also examined whether the new neurons affected by trace memory formation persist indefinitely or only as long as the hippocampus is necessary for the expression of those memories. Groups of adult rats were injected with bromodeoxyuridine (BrdU), a marker of dividing cells, and trained 1 week later with paired stimuli using a trace eyeblink conditioning task or exposed to the same number of unpaired stimuli. Cell survival was assessed after different numbers of training trials and survival periods after training. Overall cell survival was not enhanced by exposure to 200 trials of paired stimuli during trace conditioning. However, there was a positive correlation between performance of individual animals and cell survival. In addition, exposure to 800 trials of paired stimuli during trace conditioning increased the number of BrdU-labeled cells 60 d after training. The vast majority of these cells were neurons and coexpressed the neuronal markers class IIIbeta-tubulin or neuronal nuclei. These data suggest that individual differences in associative learning predict whether new neurons will survive and that once affected, these neurons remain for months and beyond the time when they are required for the retention of trace memories.


Offspring sex ratio in red-winged blackbirds is dependent on maternal age.
In a marsh-breeding population of red-winged blackbirds, the sex ratio of offspring that survived to leave the nest varied with maternal age. Old mothers produced an excess of male fledglings, middle-aged mothers produced almost equal proportions of males and females, and young mothers produced nearly twice as many females as males. More males than females hatched from the eggs of old mothers, whereas among newly hatched progeny of middle-aged and young mothers the sex ratio did not differ from unity. The hatching rate of eggs of old mothers was unusually low, suggesting that the biased sex ratio of their hatchlings may have been caused by more frequent death of female embryos, although other possibilities can be imagined. Starvation of nestlings after hatching also affected the sex ratio among young that left the nest. When starvation occurred, it fell principally on young produced by the last and next-to-last eggs laid in the clutch. Because old mothers allocated relatively more energy to those eggs than to earlier-laid eggs, whereas young mothers apportioned energy equally to their eggs, few nestlings of old mothers but many nestlings of young mothers starved. Most nestlings that died were male. It followed that the male bias in sex ratio of progeny of old mothers did not change between hatching and nestleaving, but the ratio among progeny of young mothers shifted after hatching to a strong bias favoring females at nest-leaving.


miRNA expression profiles in cerebrospinal fluid and blood of patients with acute ischemic stroke.
The aims of the study were (1) to determine whether miRNAs (microRNAs) can be detected in the cerebrospinal fluid (CSF) and blood of patients with ischemic stroke and (2) to compare these miRNA profiles with corresponding profiles from other neurological patients to address whether the miRNA profiles of CSF or blood have potential usefulness as diagnostic biomarkers of ischemic stroke. CSF from patients with acute ischemic stroke (n = 10) and patients with other neurological diseases (n = 10) was collected by lumbar puncture. Blood samples were taken immediately after. Expression profiles in the cell-free fractions of CSF and blood were analyzed by a microarray technique (miRCURY LNA™ microRNA Array, Exiqon A/S, Denmark) using a quantitative PCR (qPCR) platform containing 378 miRNA primers. In total, 183 different miRNAs were detected in the CSF, of which two miRNAs (let-7c and miR-221-3p) were found upregulated in relation to stroke. In the blood, 287 different miRNAs were detected of which two miRNAs (miR-151a-3p and miR-140-5p) were found upregulated and one miRNA (miR-18b-5p) was found downregulated in the stroke group. Some miRNAs occurred exclusively in the CSF including miR-523-3p which was detected in 50 % of the stroke patients, whereas it was completely absent in controls. Our preliminary results demonstrate that it is possible to detect and profile miRNAs in CSF and blood from patients with neurological diseases. Some miRNAs appear differentially expressed in the CSF and others in the blood of stroke patients. Currently, we are validating our results in larger groups of patients.


Effect of the delta subunit on assembly and proton permeability of the F0 proton channel of Escherichia coli F1F0 ATPase.
During the assembly of the Escherichia coli proton-translocating ATPase, the subunits of F1 interact with F0 to increase the proton permeability of the transmembrane proton channel. We tested the involvement of the delta subunit in this process by partially and completely deleting uncH (delta subunit) from a plasmid carrying the genes for the F0 subunits and delta and testing the effects of those F0 plasmids on the growth of unc+ and unc mutant E. coli strains. We found that the delta subunit was required for inhibition of growth of unc+ cells. We also tested membranes isolated from unc-deleted cells containing F0 plasmids for F1-binding ability. In unc-deleted cells, these plasmids produced F0 in amounts comparable to those found in normal unc+ E. coli cells, while having only small effects on cell growth. These studies demonstrate that the delta subunit plays an important role in opening the F0 proton channel but that it does not serve as a temporary plug of F0 during assembly, as had been previously speculated (S. Pati and W. S. A. Brusilow, J. Biol. Chem. 264:2640-2644, 1989).


Preservation of cortical histamine H3 receptors in ischemic vascular and mixed dementias.
Histamine H(3) receptor antagonists have been proposed as a novel therapeutic approach for the symptomatic treatment of Alzheimer's disease (AD). However, it is unclear whether there is a neurochemical basis for extending their potential use in vascular and mixed dementias. In this study, we measured cortical H(3) receptors in patients with subcortical ischemic vascular dementia (SIVD) and mixed SIVD/AD (MIX). Radioligand binding assays using [(3)H]GSK189254 were used to measure H(3) receptors in the postmortem frontal cortex, anterior cingulate gyrus and hippocampus of a cohort of longitudinally assessed SIVD, MIX and age-matched controls. H(3) receptor levels were unchanged in SIVD and MIX in all areas studied. Furthermore, frontal H(3) receptor densities negatively correlated with predeath assessment of cognition using Mini-Mental State Examination (MMSE) scores. Our data suggest that H(3) receptors are preserved in SIVD and MIX, thus supporting further assessments of H(3) antagonists as potential therapeutics in these dementias.


Effect of Ho dopant on the ferromagnetic characteristics of MoS2 nanocrystals.
Transition-metal dichalcogenides (TMDs) have been widely studied for decades in the spintronic device application. However, designing and tuning the magnetic properties of molybdenum disulphide (MoS2) is still a great challenge. In this work, the as-synthesized MoS2:Ho3+ nanocrystals exhibit robust room-temperature ferromagnetism and the ferromagnetism is controlled by different doping concentrations of Ho3+ ions. The maximum saturation magnetization is 0.055 emu g-1. Based on first-principles calculations, the intrinsic ferromagnetic and the changed electronic properties are introduced by Ho3+ doping. The as-obtained MoS2:Ho3+ nanocrystals with intrinsic ferromagnetism may be a potential candidate in the spintronics field and this approach may be employed in the magnetic applications of other TMDs.


Expanded adipose-derived stem cells for the treatment of complex perianal fistula: a phase II clinical trial.
The feasibility and safety of stem cell-based therapy with expanded adipose-derived stem cells (ASCs) has been investigated in a phase I clinical trial. The present study was designed as a phase II multicenter, randomized controlled trial to further investigate the effectiveness and safety of ASCs in the treatment of complex perianal fistulas. Patients with complex perianal fistulas (cryptoglandular origin, n = 35; associated with Crohn's disease, n = 14) were randomly assigned to intralesional treatment with fibrin glue or fibrin glue plus 20 million ASCs. Fistula healing and quality of life (SF-12 questionnaire) were evaluated at eight weeks and one year. If healing was not seen at eight weeks, a second dose of fibrin glue or fibrin glue plus 40 million ASCs was administered. Fistula healing was observed in 17 (71 percent) of 24 patients who received ASCs in addition to fibrin glue compared with 4 (16 percent) of 25 patients who received fibrin glue alone (relative risk for healing, 4.43; confidence interval, 1.74-11.27); P < 0.001). The proportion of patients with healing was similar in Crohn's and non-Crohn's subgroups. ASCs were also more effective than fibrin glue alone in patients with a suprasphincteric fistulous tract (P = 0.001). Quality of life scores were higher in patients who received ASCs than in those who received fibrin glue alone. At one year follow-up, the recurrence rate in patients treated with ASCs was 17.6 percent. Both treatments were well tolerated. Administration of expanded ASCs (20 to 60 million cells) in combination with fibrin glue is an effective and safe treatment for complex perianal fistula and appears to achieve higher rates of healing than fibrin glue alone.


microRNA machinery is an integral component of drug-induced transcription inhibition in HIV-1 infection.
RNA interference plays a significant role in manipulating cellular and viral mechanisms to maintain latency during HIV-1 infection. HIV-1 produces several microRNAs including one from the TAR element which alter the host's response to infection. Since cyclin/cdk complexes are important for viral transcription, these studies focus on the possible cdk inhibitors that inhibit viral transcription, without affecting normal cellular mechanisms. Roscovitine and Flavopiridol are well-studied cdk inhibitors that are effective at suppressing their target cdks at a low IC50. These cdk inhibitors and possibly future generations of drugs are affected by microRNA mechanisms. From our studies, we developed a third generation derivative called CR8#13. In cells that lack Dicer there was a higher level of basal viral LTR-reporter transcription. When drugs, specifically Flavopiridol and CR8#13 were added, the transcriptional inhibition of the LTR was less potent in cells that lacked Dicer. Also, after transfection with HIV-1 clone (pNL4.3), CR8 and CR8#13 derivatives were shown to be more effective viral transcription inhibitors in cell lines that contained Dicer (T-cells) as compared to Dicer deficient lines (monocytes). We next asked whether the addition of CR8 or CR8#13 could possibly increase levels of TAR microRNA in HIV-1 LTR containing cells. We demonstrate that the 3'TAR microRNA is produced in higher amounts after drug treatment, resulting in microRNA recruitment to the LTR. MicroRNA recruitment results in chromatin alteration, changes in Pol II phosphorylation and viral transcription inhibition. In conclusion, our results indicate that viral microRNA, specifically the TAR microRNA produced from the HIV-1 LTR is responsible for maintaining latent infections by manipulating host cell mechanisms to limit transcription from the viral LTR promoter. With the microRNA machinery present, cdk inhibitors are able to significantly increase the amount of TAR microRNA, leading to downregulation of viral LTR transcription.


Sacred moments: implications on well-being and stress.
This research provides a theoretical, empirical, and qualitative examination of the role of cultivating sacred moments in daily life on subjective well-being (SWB), psychological well-being (PWB), and stress. Seventy-three participants were randomly assigned to two groups: (a) a 3-week intervention group where members were instructed in cultivating sacred moments, or (b) a 3-week control group where members were instructed in writing about daily activities. Findings indicate that the intervention was equally as effective as an adapted therapeutic writing intervention. There were significant effects over time across multiple assessments related to SWB, PWB, stress, and daily spiritual experiences after the 3-week intervention and again 6 weeks later. Qualitative analysis complemented and enriched the findings of these results. This study introduces a new intervention into the field of clinical psychology and extends the findings of prior research.


Endogenous IL-17 as a factor determining the severity of Clostridium difficile infection in mice.
Clostridium difficile infection (CDI) is a toxin-mediated intestinal disease. Toxin A, toxin B and binary toxin are believed to be responsible for the pathogenesis of CDI, which is characterized by massive infiltration of neutrophils at the infected intestinal mucosa. IL-17 is one of the cytokines that play critical roles in several inflammatory and immunological diseases through various actions, including promoting neutrophil recruitment. The aim of this study was to examine the role of this cytokine in CDI by employing IL-17 A and F double knockout (IL-17 KO) mice for the CDI model. We demonstrated that IL-17 KO mice were more resistant to CDI than WT mice using several factors, such as diarrhoea score, weight change and survival rate. Although the bacterial numbers of C. difficile in faeces were not different, the inflammatory mediator levels at the large intestine on day 3 post-infection were attenuated in IL-17 KO mice. Finally, we showed that infiltration of neutrophils, but not macrophages, in the large intestine was significantly decreased in IL-17 KO mice compared to WT mice. In conclusion, the data demonstrate that endogenous IL-17 may be a factor determining the severity of CDI in mice. Although the mechanism is totally unknown, IL-17-mediated inflammatory responses, such as cytokine/chemokine production and neutrophil accumulation, may be plausible targets for future investigations.


Leaf and Plant Age Affects Photosynthetic Performance and Photoprotective Capacity.
In this work, we studied the changes in high-light tolerance and photosynthetic activity in leaves of the Arabidopsis (Arabidopsis thaliana) rosette throughout the vegetative stage of growth. We implemented an image-analysis work flow to analyze the capacity of both the whole plant and individual leaves to cope with excess excitation energy by following the changes in absorbed light energy partitioning. The data show that leaf and plant age are both important factors influencing the fate of excitation energy. During the dark-to-light transition, the age of the plant affects mostly steady-state levels of photochemical and nonphotochemical quenching, leading to an increased photosynthetic performance of its leaves. The age of the leaf affects the induction kinetics of nonphotochemical quenching. These observations were confirmed using model selection procedures. We further investigated how different leaves on a rosette acclimate to high light and show that younger leaves are less prone to photoinhibition than older leaves. Our results stress that both plant and leaf age should be taken into consideration during the quantification of photosynthetic and photoprotective traits to produce repeatable and reliable results.


Sleepwalking, REM sleep behaviour disorder and overlap parasomnia in patients with Parkinson's disease.
In a questionnaire survey, we identified 36 (9%) of 417 Parkinson's disease (PD) patients with sleepwalking (SW); 72% of them also had a history of REM sleep behaviour disorder (RBD). We aimed to assess the clinical and polysomnographic characteristics of SW in PD and to compare them to patients with PD with and without a history of RBD. We performed video-polysomnography and detailed clinical examination in 30 PD patients from the above-mentioned survey: 10 patients with a history of SW, 10 patients with a history of RBD, and 10 patients with no history of either SW or RBD. PD patients with SW had higher depression, anxiety and Hoehn & Yahr scores and lower activities of daily living scores than patients without a history of RBD but did not differ from patients with RBD. Patients with SW and RBD also had more often dyskinesia and hallucinations. By polysomnography, RBD was observed in 8 patients with SW and in all patients with a history of RBD. A total of 5 patients without a history of either SW or RBD had REM sleep without atonia without behavioural peculiarities. SW in PD is associated with depression, higher disease severity and functional disability. The simultaneous occurrence of SW and RBD (overlap parasomnia) in most patients suggests a common underlying disturbance of motor control during sleep in PD, with variable manifestations in different sleep stages.


Food-dependent exercise-induced anaphylaxis with negative allergy testing.
Food-dependent exercise-induced anaphylaxis (FDEIA) is a disorder where exercise following allergen ingestion triggers anaphylaxis although exercise and allergen exposure are independently tolerated. The diagnosis of FDEIA is based on a characteristic clinical history. The culprit allergen is usually confirmed through the use of skin prick testing (SPT) serum-specific IgE levels and a food-exercise challenge. We present a case of FDEIA suggested by clinical history and open food-exercise challenge with negative specific IgE levels and SPT that highlights the challenges involved in diagnosing and managing this rare disorder.


Patterns of Growth and Decline in Lung Function in Persistent Childhood Asthma.
Tracking longitudinal measurements of growth and decline in lung function in patients with persistent childhood asthma may reveal links between asthma and subsequent chronic airflow obstruction. We classified children with asthma according to four characteristic patterns of lung-function growth and decline on the basis of graphs showing forced expiratory volume in 1 second (FEV1), representing spirometric measurements performed from childhood into adulthood. Risk factors associated with abnormal patterns were also examined. To define normal values, we used FEV1 values from participants in the National Health and Nutrition Examination Survey who did not have asthma. Of the 684 study participants, 170 (25%) had a normal pattern of lung-function growth without early decline, and 514 (75%) had abnormal patterns: 176 (26%) had reduced growth and an early decline, 160 (23%) had reduced growth only, and 178 (26%) had normal growth and an early decline. Lower baseline values for FEV1, smaller bronchodilator response, airway hyperresponsiveness at baseline, and male sex were associated with reduced growth (P<0.001 for all comparisons). At the last spirometric measurement (mean [±SD] age, 26.0±1.8 years), 73 participants (11%) met Global Initiative for Chronic Obstructive Lung Disease spirometric criteria for lung-function impairment that was consistent with chronic obstructive pulmonary disease (COPD); these participants were more likely to have a reduced pattern of growth than a normal pattern (18% vs. 3%, P<0.001). Childhood impairment of lung function and male sex were the most significant predictors of abnormal longitudinal patterns of lung-function growth and decline. Children with persistent asthma and reduced growth of lung function are at increased risk for fixed airflow obstruction and possibly COPD in early adulthood. (Funded by the Parker B. Francis Foundation and others; ClinicalTrials.gov number, NCT00000575.).