Strategies and Challenges: Qualitative Lessons Learned From Georgia Produce Prescription Programs.
Produce prescription programs (PPPs) have grown in numbers in the past decade, empowering health care providers to promote health by issuing subsidies for produce to vulnerable patients. However, little research has been conducted on the facilitators that make it easier for PPPs to succeed or the barriers that programs face, which could provide guidance on how to improve future PPP design and implementation. The study sought to identify the facilitators and barriers affecting positive outcomes in Georgia PPPs called Fruit and Vegetable Prescription (FVRx) Programs. A process evaluation with a qualitative comparative case study approach was conducted. Fifteen FVRx providers, ranging from nutrition educators to farmers market managers, were interviewed in a focus group interview or on the phone between 2016 and 2017. Two nutrition education classes and an FVRx best practices meeting were observed, and program documents were collected. Interview transcripts, field notes from observations, and documents were then thematically analyzed. Four overall themes were determined regarding facilitators and barriers experienced by FVRx programs: (1) creating accessible programming may encourage FVRx participation, (2) provider dedication to the program is important, (3) participants' challenging life circumstances can make participation difficult, and (4) the sustainability of the program is a concern. The findings of this study suggest helpful strategies and challenges for providers to consider when developing and implementing PPPs in Georgia and beyond. Research on the long-term program impact is needed, and policy options for sustainable, scaling up of PPPs should be explored.


Characterization of the major light-harvesting complexes (LHCBM) of the green alga Chlamydomonas reinhardtii.
Nine genes (LHCBM1-9) encode the major light-harvesting system of Chlamydomonas reinhardtii. Transcriptomic and proteomic analyses have shown that those genes are all expressed albeit in different amounts and some of them only in certain conditions. However, little is known about the properties and specific functions of the individual gene products because they have never been isolated. Here we have purified several complexes from native membranes and/or we have reconstituted them in vitro with pigments extracted from C. reinhardtii. It is shown that LHCBM1 and -M2/7 represent more than half of the LHCBM population in the membrane. LHCBM2/7 forms homotrimers while LHCBM1 seems to be present in heterotrimers. Trimers containing only type I LHCBM (M3/4/6/8/9) were also observed. Despite their different roles, all complexes have very similar properties in terms of pigment content, organization, stability, absorption, fluorescence and excited-state lifetimes. Thus the involvement of LHCBM1 in non-photochemical quenching is suggested to be due to specific interactions with other components of the membrane and not to the inherent quenching properties of the complex. Similarly, the overexpression of LHCBM9 during sulfur deprivation can be explained by its low sulfur content as compared with the other LHCBMs. Considering the highly conserved biochemical and spectroscopic properties, the major difference between the complexes may be in their capacity to interact with other components of the thylakoid membrane.


Renal Delivery of Anti-microRNA Oligonucleotides in Rats.
MicroRNAs are endogenous small, non-protein-coding RNA molecules that play an important role in the regulation of a wide variety of cellular functions and disease processes. A novel role for microRNAs in the development of hypertension and hypertensive tissue injury is emerging in recent studies. Development of hypertension involves multiple organ systems and cannot be modeled in vitro. Therefore, the ability to experimentally alter genes, gene products, or biological pathways, including microRNAs, in an organ-specific manner in intact animal models is particularly valuable to hypertension research. The kidney plays a central role in the long-term regulation of arterial blood pressure. In this chapter, we describe a detailed protocol for using a renal interstitial injection method to deliver anti-miR oligonucleotides to knock down microRNA specifically in the kidney in conscious rats.


Ultrasound in prenatal diagnosis: polemics around routine ultrasound screening for second trimester fetal malformations.
Ultrasound for routine fetal malformation screening has been polemical from its early beginning because of the very broad range of diagnosis rates disclosed, i.e. from 13% to 82%, average 27.5%. A review of available studies is proposed to assess objectively the efficacy of ultrasound, considering also economical, ethical and methodological aspects as influential factors for choosing a routine screening policy. The utility of fetal malformation diagnosis before birth is brought forward, including second opinion, karyotyping, poly-disciplinary case discussion prior to management. Method and material of reviewed studies considerably vary and might influence the sensitivity results, as the choice of the population sample and selection of pregnant women, gestation age at screening, distribution of malformation among systems or tracts, exclusion of some fetal malformation and the routine practice of autopsy. Efficiency of screening studies is compared, and among them Radius and Eurofetus studies. Average sensitivity is finally considered as satisfactory in the daily practice when operated by trained personnel. The importance of additional factors for successful screening are emphasized such as education, equipment quality and fetal ultrasound examination at different gestation age for a better understanding of natural history of fetal morphology.


Membraneless water filtration using CO2.
Water purification technologies such as microfiltration/ultrafiltration and reverse osmosis utilize porous membranes to remove suspended particles and solutes. These membranes, however, cause many drawbacks such as a high pumping cost and a need for periodic replacement due to fouling. Here we show an alternative membraneless method for separating suspended particles by exposing the colloidal suspension to CO2. Dissolution of CO2 into the suspension creates solute gradients that drive phoretic motion of particles. Due to the large diffusion potential generated by the dissociation of carbonic acid, colloidal particles move either away from or towards the gas-liquid interface depending on their surface charge. Using the directed motion of particles induced by exposure to CO2, we demonstrate a scalable, continuous flow, membraneless particle filtration process that exhibits low energy consumption, three orders of magnitude lower than conventional microfiltration/ultrafiltration processes, and is essentially free from fouling.


Exhaled Breath Markers for Nonimaging and Noninvasive Measures for Detection of Multiple Sclerosis.
Multiple sclerosis (MS) is the most common chronic neurological disease affecting young adults. MS diagnosis is based on clinical characteristics and confirmed by examination of the cerebrospinal fluids (CSF) or by magnetic resonance imaging (MRI) of the brain or spinal cord or both. However, neither of the current diagnostic procedures are adequate as a routine tool to determine disease state. Thus, diagnostic biomarkers are needed. In the current study, a novel approach that could meet these expectations is presented. The approach is based on noninvasive analysis of volatile organic compounds (VOCs) in breath. Exhaled breath was collected from 204 participants, 146 MS and 58 healthy control individuals. Analysis was performed by gas-chromatography mass-spectrometry (GC-MS) and nanomaterial-based sensor array. Predictive models were derived from the sensors, using artificial neural networks (ANNs). GC-MS analysis revealed significant differences in VOC abundance between MS patients and controls. Sensor data analysis on training sets was able to discriminate in binary comparisons between MS patients and controls with accuracies up to 90%. Blinded sets showed 95% positive predictive value (PPV) between MS-remission and control, 100% sensitivity with 100% negative predictive value (NPV) between MS not-treated (NT) and control, and 86% NPV between relapse and control. Possible links between VOC biomarkers and the MS pathogenesis were established. Preliminary results suggest the applicability of a new nanotechnology-based method for MS diagnostics.


Evaluation of vancomycin use in university-affiliated hospitals in Southern Khorasan Province (East Iran) based on HICPAC guidelines.
Vancomycin resistance has raised concerns about its effectiveness prospect in the treatment of patients with Gram-positive infections. The Healthcare Infection Control Practices Advisory Committee (HICPAC) has recently established guidelines to delineate improper use of vancomycin. In this light, we sought out to determine the appropriateness of vancomycin prescription using the HICPAC guidelines. The study was carried out in two university-affiliated hospitals, Valiasr and Imam Reza, with 297 and 234 beds, respectively, from May 2012 to May 2013. This retrospective study evaluated the vancomycin prescription and usage in the hospitals. Total vancomycin use was determined and expressed as vancomycin courses per 298 admitted patients. The patient information was collected on a data collection sheet as follows: demographic variables, etiology and localization of infection, microbiological data, duration of vancomycin treatment, reasons for vancomycin prescription, prescribed antibiotic dosing, and patient regimen. The average age of the patients and vancomycin treatment duration were 55.965 years and 10.5 days, respectively. Septicemia (15.7%) was the most common cause of vancomycin administration. Vancomycin use was documented to be appropriate and inappropriate in 236 (89.4%) and 28 (10.6%) patients, respectively. No statistically significant differences were found among the wards and hospitals (P values =0.66 and 0.54, respectively) in terms of appropriateness of vancomycin use based on the HICPAC criteria. In addition, 29.21% and 62% of all patients exhibited complete and partial recovery, respectively. We found that 90% of the cases showed compliance with the HICPAC recommendations. Comprehensive programs are required to improve the vancomycin use in the hospitals. Vancomycin use should be monitored due to its large-scale empiric use. The rate of improper use of vancomycin in the infection and intensive care unit services may be high, and pharmacists must take appropriate action to optimize the use of the drug.


Hemotin, a Regulator of Phagocytosis Encoded by a Small ORF and Conserved across Metazoans.
Translation of hundreds of small ORFs (smORFs) of less than 100 amino acids has recently been revealed in vertebrates and Drosophila. Some of these peptides have essential and conserved cellular functions. In Drosophila, we have predicted a particular smORF class encoding ~80 aa hydrophobic peptides, which may function in membranes and cell organelles. Here, we characterise hemotin, a gene encoding an 88aa transmembrane smORF peptide localised to early endosomes in Drosophila macrophages. hemotin regulates endosomal maturation during phagocytosis by repressing the cooperation of 14-3-3ζ with specific phosphatidylinositol (PI) enzymes. hemotin mutants accumulate undigested phagocytic material inside enlarged endo-lysosomes and as a result, hemotin mutants have reduced ability to fight bacteria, and hence, have severely reduced life span and resistance to infections. We identify Stannin, a peptide involved in organometallic toxicity, as the Hemotin functional homologue in vertebrates, showing that this novel regulator of phagocytic processing is widely conserved, emphasizing the significance of smORF peptides in cell biology and disease.


[Effectiveness of vagotomy after repeated operations in duodenal ulcer].
The results of surgical therapy in 24 patients in whom reoperations on the stomach were supplemented with vagotomy are analysed. It is felt that in complicated peptic ulcers of gastroenteroanastomosis in operations after suturing perforated duodenal ulcer, in erosive gastritis of the gastric stump with hemorrhages reoperations on the stomach should be associated with vagotomy.


Surface zone articular chondrocytes modulate the bulk and surface mechanical properties of the tissue-engineered cartilage.
The central hypothesis of functional tissue engineering is that an engineered construct can serve as a viable replacement tissue in vivo by replicating the structure and function of native tissue. In the case of articular cartilage, this requires the reproduction of the bulk mechanical and surface lubrication properties of native hyaline cartilage. Cartilage tissue engineering has primarily focused on achieving the bulk mechanical properties of native cartilage such as the compressive aggregate modulus and tensile strength. A scaffold-free self-assembling process has been developed that produces engineered cartilage with compressive properties approaching native tissue levels. Thus, the next step in this process is to begin addressing the friction coefficient and wear properties of these engineered constructs. The superficial zone protein (SZP), also known as lubricin or PRG4, is a boundary mode lubricant that is synthesized by surface zone (SZ) articular chondrocytes. Under conditions of high loading and low sliding speeds, SZP reduces friction and wear at the articular surface. The objective of this investigation was to determine whether increasing the proportion of SZ chondrocytes in cartilage constructs, in the absence of external stimuli such as growth factors and mechanical loading, would enhance the secretion of SZP and improve their frictional properties. In this study, cartilage constructs were engineered through a self-assembling process with varying ratios of SZ and middle zone (MZ) chondrocytes (SZ:MZ): 0:100, 25:75, 50:50, 75:25, and 100:0. Constructs containing different ratios of SZ and MZ chondrocytes did not significantly differ in the glycosaminoglycan composition or compressive aggregate modulus. In contrast, tensile properties and collagen content were enhanced in nearly all constructs containing greater amounts of SZ chondrocytes. Increasing the proportion of SZ chondrocytes had the hypothesized effect of improving the synthesis and secretion of SZP. However, increasing the SZ chondrocyte fraction did not significantly reduce the friction coefficient. These results demonstrate that additional factors, such as SZP-binding macromolecules, surface roughness, and adhesion, need to be examined to modulate the lubrication properties of engineered cartilage.


Up-Regulation of Oligodendrocyte Lineage Markers in the Cerebellum of Autistic Patients: Evidence from Network Analysis of Gene Expression.
Autism is a neurodevelopmental disorder manifested by impaired social interaction, deficits in communication skills, restricted interests, and repetitive behaviors. In neurodevelopmental, neurodegenerative, and psychiatric disorders, glial cells undergo morphological, biochemical, and functional rearrangements, which are critical for neuronal development, neurotransmission, and synaptic connectivity. Cerebellar function is not limited to motor coordination but also contributes to cognition and may be affected in autism. Oligodendrocytes and specifically oligodendroglial precursors are highly susceptible to oxidative stress and excitotoxic insult. In the present study, we searched for evidence for developmental oligodendropathy in the context of autism by performing a network analysis of gene expression of cerebellar tissue. We created an in silico network model (OLIGO) showing the landscape of interactions between oligodendrocyte markers and demonstrated that more than 50 % (16 out of 30) of the genes within this model displayed significant changes of expression (corrected p value <0.05) in the cerebellum of autistic patients. In particular, we found up-regulation of OLIG2-, MBP-, OLIG1-, and MAG-specific oligodendrocyte markers. We postulate that aberrant expression of oligodendrocyte-specific genes, potentially related to changes in oligodendrogenesis, may contribute to abnormal cerebellar development, impaired myelination, and anomalous synaptic connectivity in autism spectrum disorders (ASD).


Oxygen activation at a dicobalt centre of a dipyridylethane naphthyridine complex.
The mechanism of oxygen activation at a dicobalt bis-μ-hydroxo core is probed by the implementation of synthetic methods to isolate reaction intermediates. Reduction of a dicobalt(iii,iii) core ligated by the polypyridyl ligand dipyridylethane naphthyridine (DPEN) by two electrons and subsequent protonation result in the release of one water moiety to furnish a dicobalt(ii,ii) center with an open binding site. This reduced core may be independently isolated by chemical reduction. Variable-temperature 1H NMR and SQUID magnetometry reveal the reduced dicobalt(ii,ii) intermediate to consist of two low spin Co(ii) centers coupled antiferromagnetically. Binding of O2 to the open coordination site of the dicobalt(ii,ii) core results in the production of an oxygen adduct, which is proposed to be a dicobalt(iii,iii) peroxo. Electrochemical studies show that the addition of two electrons results in cleavage of the O-O bond.


Microplastics Contamination in the Edible Fish Mozambique Tilapia (Oreochromis mossambicus) from the Selvampathy Wetland of Coimbatore, Tamil Nadu, India.
The present study investigated the microplastics (MPs) contamination in the gastrointestinal (GI) tract, gills and muscle of the Mozambique tilapia, Oreochromis mossambicus sampled from the Selvampathy Lake of Coimbatore, Tamil Nadu. MPs abundance was found in 10 to 28, 8 to 27, and 4 to 12 particles and their size ranged between 4.4 and 210, 4.6 to 180, and 4.5 to 194 μm in the GI tract, gills and muscle, respectively. MPs were dominantly shaped as fibres (95%) and fragments (5%) with the following colour pattern of blue > black > pink > transparent > and others. Extracted MPs polymer nature were polyethylene (54%), polyamide (38%) and polypropylene (8%). The present study reveals that the edible fish O. mossambicus had MPs that can be transferred to consumers. Moreover, urban discharges, including domestic wastes, agricultural and rainwater runoff, might be possible MPs sources to the studied wetland.


SVM feature selection based rotation forest ensemble classifiers to improve computer-aided diagnosis of Parkinson disease.
Parkinson disease (PD) is an age-related deterioration of certain nerve systems, which affects movement, balance, and muscle control of clients. PD is one of the common diseases which affect 1% of people older than 60 years. A new classification scheme based on support vector machine (SVM) selected features to train rotation forest (RF) ensemble classifiers is presented for improving diagnosis of PD. The dataset contains records of voice measurements from 31 people, 23 with PD and each record in the dataset is defined with 22 features. The diagnosis model first makes use of a linear SVM to select ten most relevant features from 22. As a second step of the classification model, six different classifiers are trained with the subset of features. Subsequently, at the third step, the accuracies of classifiers are improved by the utilization of RF ensemble classification strategy. The results of the experiments are evaluated using three metrics; classification accuracy (ACC), Kappa Error (KE) and Area under the Receiver Operating Characteristic (ROC) Curve (AUC). Performance measures of two base classifiers, i.e. KStar and IBk, demonstrated an apparent increase in PD diagnosis accuracy compared to similar studies in literature. After all, application of RF ensemble classification scheme improved PD diagnosis in 5 of 6 classifiers significantly. We, numerically, obtained about 97% accuracy in RF ensemble of IBk (a K-Nearest Neighbor variant) algorithm, which is a quite high performance for Parkinson disease diagnosis.


[Reliability of the WPPSI-R test in the evaluation of cognitive development in preschool children].
The WPPSI-R scale (Wechsler Preschool and Primary Scale of Intelligence - Revised) is a psychometric test chosen as the evaluation tool in a study on preschool-age cognitive development in a cohort of very low birth weight (VLBW) premature children from the Fernandes Figueira Institute (IFF), applied by four previously trained psychologists. The objective of this study was to verify inter-observer reliability in the test application. Two types of reliability study design were used: balanced incomplete blocks, to verify agreement in the application of the scale, and crossed design, to verify agreement in scoring of items. We studied 12 preschool children born at IFF (birthweight < 1,500g). The intraclass correlation coefficients (ICC) were: 0.82 (full-scale IQ), 0.89 (verbal IQ), and 0.91 (performance IQ), in the incomplete block design study, and 0.99, 0.98, and 0.99, respectively, in the crossed design study, indicating good reliability. Application of the WPPSI-R scale in the study of cognitive development of VLBW premature children at IFF proved adequate, as shown by these results.


A Novel Registration-Based Semiautomatic Mandible Segmentation Pipeline Using Computed Tomography Images to Study Mandibular Development.
We present a registration-based semiautomatic mandible segmentation (SAMS) pipeline designed to process a large number of computed tomography studies to segment 3-dimensional mandibles. The pipeline consists of a manual preprocessing step, an automatic segmentation step, and a final manual postprocessing step. The automatic portion uses a nonlinear diffeomorphic method to register each preprocessed input computed tomography test scan on 54 reference templates, ranging in age from birth to 19 years. This creates 54 segmentations, which are then combined into a single composite mandible. This pipeline was assessed using 20 mandibles from computed tomography studies with ages 1 to 19 years, segmented using both SAMS-processing and manual segmentation. Comparisons between the SAMS-processed and manually-segmented mandibles revealed 97% similarity agreement with comparable volumes. The resulting 3-dimensional mandibles were further enhanced with manual postprocessing in specific regions. Findings are indicative of a robust pipeline that reduces manual segmentation time by 75% and increases the feasibility of large-scale mandibular growth studies.


The role of organic matter lost in kraft pulping material balances .
Efficiency improvement in a pulp mill includes minimisation of environmental discharges simultaneously with the development of pulp quality and production economy. Material balances in production processes, including fate of sidestreams, are key in proceeding these matters. Different approaches of determining the material balances increase understanding of process behaviour. We have focused on measuring sidestream (carryovers, washing water, filtrate) dissolved organic matter (DOM) in fibreline unit process blocks of softwood ECF bleached kraft production. The DOM was analysed by traditional wastewater methods (volatile solids, organic carbon, chemical oxygen demand). The measured data was combined with primarily simulated water balances and routine operational mill data in a simulation model. From this balance, yield estimate included, lost organic matter through complete degradation (CD) and volatile organic compounds (VOC) can be calculated throughout the fibreline. The sensitivity of this considerable amount (23-35 kgDVS/adt in total) to various factors is discussed in this paper.


Symptomatic and preventive medication use according to age and frailty in Australian and Japanese nursing homes.
To investigate symptomatic and preventive medication use according to age and frailty in Australian and Japanese nursing homes (NHs). Secondary cross-sectional analyses of two prospective cohort studies involving 12 Australian NHs and four Japanese NHs. Frailty was measured using the FRAIL-NH scale (non-frail 0-2; frail 3-6; most-frail 7-14). Regular medications were classified as symptomatic or preventive based on published lists and expert consensus. Descriptive statistics were used to compare the prevalence and ratio of symptomatic to preventive medications. Overall, 550 Australian residents (87.7 ± 7.3 years; 73.3% females) and 333 Japanese residents (86.5 ± 7.0 years; 73.3% females) were included. Australian residents used a higher mean number of medications than Japanese residents (9.8 ± 4.0 vs 7.7 ± 3.7, p < 0.0001). Australian residents used more preventive than symptomatic medications (5.5 ± 2.5 vs 4.3 ± 2.6, p < 0.0001), while Japanese residents used more symptomatic than preventive medications (4.7 ± 2.6 vs 3.0 ± 2.2, p < 0.0001). In Australia, symptomatic medications were more prevalent with increasing frailty (non-frail 3.4 ± 2.6; frail 4.0 ± 2.6; most-frail 4.8 ± 2.6, p < 0.0001) but less prevalent with age (< 80 years 5.0 ± 2.9; 80-89 years 4.4 ± 2.6; ≥ 90 years 3.9 ± 2.5, p = 0.0042); while preventive medications remained similar across age and frailty groups. In Japan, there was no significant difference in the mean number of symptomatic and preventive medications irrespective of age and frailty. The ratio of symptomatic to preventive medications was higher with increasing frailty but lower with age in Australia; whereas in Japan, the ratio remained consistent across age and frailty groups. Preventive medications remained prevalent in most-frail residents in both cohorts, albeit at lower levels in Japan.


Interleukin-2-receptor blockade with daclizumab to prevent acute rejection in renal transplantation. Daclizumab Triple Therapy Study Group.
Monoclonal antibodies that block the high-affinity interleukin-2 receptor expressed on alloantigen-reactive T lymphocytes may cause selective immunosuppression. Daclizumab is a genetically engineered human IgG1 monoclonal antibody that binds specifically to the alpha chain of the interleukin-2 receptor and may thus reduce the risk of rejection after renal transplantation. We administered daclizumab (1.0 mg per kilogram of body weight) or placebo intravenously before transplantation and once every other week afterward, for a total of five doses, to 260 patients receiving first cadaveric kidney grafts and immunosuppressive therapy with cyclosporine, azathioprine, and prednisone. The patients were followed at regular intervals for 12 months. The primary end point was the incidence of biopsy-confirmed acute rejection within six months after transplantation. Of the 126 patients given daclizumab, 28 (22 percent) had biopsy-confirmed episodes of acute rejection, as compared with 47 of the 134 patients (35 percent) who received placebo (P=0.03). Graft survival at 12 months was 95 percent in the daclizumab-treated patients, as compared with 90 percent in the patients given placebo (P=0.08). The patients given daclizumab did not have any adverse reactions to the drug, and at six months, there were no significant differences between the two groups with respect to infectious complications or cancers. The serum half-life of daclizumab was 20 days, and its administration resulted in prolonged saturation of interleukin-2alpha receptors on circulating lymphocytes. Daclizumab reduces the frequency of acute rejection in kidney-transplant recipients.


Which symptoms and clinical features correctly identify serious respiratory infection in children attending a paediatric assessment unit?
Parent-reported symptoms are frequently used to triage children, but little is known about which symptoms identify children with serious respiratory infections. The authors aimed to identify symptoms and triage findings predictive of serious respiratory infection, and to quantify agreement between parent and nurse assessment. Prospective diagnostic cohort study. Paediatric Assessment Unit, University Hospitals Coventry and Warwickshire NHS Trust. 535 children aged between 3 months and 12 years with suspected acute infection. Parents completed a symptom questionnaire on arrival. Children were triaged by a nurse, who measured routine vital signs. The final diagnosis at discharge was used as the outcome. Symptoms and triage findings were analysed to identify features diagnostic of serious respiratory infection. Agreement between parent and triage nurse assessment was measured and kappa values calculated. Parent-reported symptoms were poor indicators of serious respiratory infection (positive likelihood ratio (LR+) 0.56-1.93) and agreed poorly with nurse assessment (kappa 0.22-0.56). The best predictor was clinical assessment of respiratory distress (LR+ 5.04). Oxygen saturations <94% were highly specific (specificity 95.1%) but had poor sensitivity (35.6%). Tachypnoea (defined by current Advanced Paediatric Life Support standards) offered little discriminatory value. Parent-reported symptoms were unreliable discriminators of serious respiratory infection in children with suspected acute infection, and did not correlate well with nurse assessment. Using symptoms to identify higher risk children in this setting is unreliable. Nurse triage assessment of respiratory distress and some vital signs are important predictors.


The inhibition of apoptosis in myositis and in normal muscle cells.
The mechanism of injury and death of muscle cells in the inflammatory myopathies (dermatomyositis, polymyositis, and inclusion body myositis) remains obscure. We and others have not detected apoptosis in the muscle biopsies from patients with myositis despite clear evidence of cell damage and loss. We provide evidence in this study that Fas ligand (FasL) as well as Fas is present on muscle cells and inflammatory cells in myositis biopsies: Fas is present on most muscle cells and lymphocytes, and FasL is present on degenerating muscle cells and many infiltrating mononuclear cells. The expression of both Fas and FasL in the inflamed tissue makes the absence of apoptosis more striking. To address the mechanisms of this resistance to classical apoptosis in muscle cells, we have investigated the expression of the antiapoptotic molecule FLICE (Fas-associated death domain-like IL-1-converting enzyme)-inhibitory protein (FLIP) in muscle biopsies of myositis patients and in cultured human skeletal muscle cells. Using laser capture microscopy, we have shown that FLIP is expressed in the muscle fibers and on infiltrating lymphocytes of myositis biopsies. Furthermore, we have shown that FLIP, but not Bcl-2, is expressed in cultured human skeletal muscle cells stimulated with proinflammatory cytokines, and inhibition of FLIP with antisense oligonucleotides promotes significant cleavage of poly(ADP-ribose) polymerase autoantigen, a sensitive indicator of apoptosis. These studies strongly suggest that the resistance of muscle to Fas-mediated apoptosis is due to the expression of FLIP in muscle cells in the inflammatory environment in myositis.


Primary closure and continuous irrigation of the perineal wound after proctectomy.
The aim was to assess the success of primary closure and continuous irrigation of the perineal wound in achieving wound healing after proctectomy. The frequency and degree of primary healing (dry wound within 30 days after removal of irrigation catheters) and the factors affecting the rate of healing were determined in 152 patients undergoing proctectomy for carcinoma (31 patients), Crohn's colitis (32 patients), or ulcerative colitis (89 patients). All patients were followed for at least 2 years. Primary healing occurred in 64.5% of patients, whereas it was delayed (dry wound by 90 days) or failed to occur in 30.2% and 5.3%, respectively. Of those with carcinoma, 87% had healing by 30 days, whereas only 61% and 50% of those with either ulcerative colitis or Crohn's disease, respectively, had healing in that time (P less than 0.02). Females had primary healing more often than males (70% versus 45%, P less than 0.01), and the incidence of persistent perineal sinus was greater in men (28%) than in women (13%) (P less than 0.05). Primary healing was adversely affected in men when drains exited centrally through the wound (31% healed primarily) as opposed to when they exited laterally (65% healed primarily, P less than 0.04). We conclude that primary closure with irrigation of the perineal wound is safe and provides satisfactory healing in most patients.


Modulation of root-skewing responses by KNAT1 in Arabidopsis thaliana.
The KNOTTED1 homeobox (KNOX) family transcription factors are essential for stem cell establishment and maintenance and regulate various aspects of development in all green plants. Expression patterns of the KNOX genes in the roots of plants have been reported, but their role in development remains unclear. Here we show how the KNAT1 gene is specifically involved in root skewing in Arabidopsis. The roots of two mutant alleles of KNAT1 (bp-1 and bp-5) exhibited exaggerated skewing to the right of gravity when grown on both vertical and tilted agar medium surfaces. This skewing phenotype was enhanced by treatments with low concentrations of propyzamide, and required auxin transport. The KNAT1 mutation substantially decreased basipetal auxin transport and increased auxin accumulation in the roots. Using a PIN2-GFP reporter and western blot analysis, we found that this alteration in auxin transport was accompanied by a decrease in PIN2 levels in the root tip. Decreased PIN2 expression in the mutant roots was not accompanied by reduced mRNA levels, suggesting that the KNAT1 mutations affected PIN2 expression at the posttranscriptional level. In vivo visualization of intracellular vacuolar targeting indicated that vacuolar degradation of PIN2-GFP was significantly promoted in the root tips of the bp allelic mutants. Together, these results demonstrate that KNAT1 negatively modulates root skewing, possibly by regulating auxin transport.


Developing and testing a middle-range theory of the well-being supportive physical environment of home-dwelling elderly.
The aim is to describe the development of a middle-range theory by using an inductive-deductive approach. A theory of well-being supporting physical environment of home-dwelling elderly is used as an example. The inductive-deductive theory development process is described through four different phases: (1) the creations of concepts were described inductively through concept synthesis, (2) relationships between the concepts were examined to set up a hypothetical model, (3) hypotheses were set up to verify the concepts and to test hypothetical models, and (4) the verification and presentation of the theory.


Long-term MI outcomes at hospitals with or without on-site revascularization.
Many studies have found that patients with acute myocardial infarction (AMI) who are admitted to hospitals with on-site revascularization facilities have higher rates of invasive cardiac procedures and better outcomes than patients in hospitals without such facilities. Whether such differences are due to invasive procedure rates alone or to other patient, physician, and hospital characteristics is unknown. To determine whether invasive procedural rate variations alone account for outcome differences in patients with AMI admitted to hospitals with or without on-site revascularization facilities. Retrospective, observational cohort study using linked population-based administrative data from a universal health insurance system. One hundred ninety acute care hospitals in Ontario, 9 of which offered invasive procedures. A total of 25 697 patients hospitalized with AMI between April 1, 1992, and December 31, 1993, of whom 2832 (11%) were in invasive hospitals. Mortality, recurrent cardiac hospitalizations, and emergency department visits in the 5 years following the index admission, adjusted for patient age, sex, socioeconomic status, illness severity, and index revascularization procedures; attending physician specialty; and hospital volume, teaching status, and geographical proximity to invasive-procedure centers and compared by hospital type. Patients admitted to invasive-procedure hospitals were much more likely to undergo revascularization (11.4% vs 3.2% at other hospitals; P<.001). However, many other clinical and process-related factors differed between the 2 groups. Although mortality rates were similar between the 2 institution types, the nonfatal composite 5-year event rate (ie, recurrent cardiac hospitalization and emergency department visits) was lower for patients initially admitted to invasive-procedure hospitals (71.3% vs 80.4%; unadjusted odds ratio [OR], 0.65; 95% confidence interval [CI], 0.52-0.82; P<.001). This advantage persisted after adjustment for sociodemographic and clinical factors and procedure utilization (adjusted OR, 0.68; 95% CI, 0.53-0.89; P<.001). However, the nonfatal outcome advantages of invasive-procedure hospitals were explained by their teaching status (adjusted OR, 0.98; 95% CI, 0.73-1.30; P =.87). In this sample of patients admitted with AMI, the differing outcomes of apparently similar patients treated in 2 different practice settings were explained by multiple competing factors. Researchers conducting observational studies should be cautious about attributing patient outcome differences to any single factor.


Changes in electromyogram power spectra of facial and jaw-elevator muscles during fatigue.
Endurance and changes in electromyogram (EMG) power spectra were investigated during a fatiguing static contraction at 50% of the maximum EMG amplitude in two jaw-elevator muscles (masseter and temporalis) and five facial muscles (frontalis, corrugator supercilii, zygomaticus major, orbicularis oris, and buccinator). Relatively high endurance was found in orbicularis oris, frontalis, and corrugator supercilii muscles; intermediate endurance was found in zygomaticus major, buccinator, and temporalis muscles; and low endurance was found in the masseter muscle. The last muscle showed a relatively fast linear decrease of the median frequency of the power spectrum. The other muscles showed a much slower, exponential decrease. The median frequency appeared to reflect reliably the changes in the shape of the power spectra during fatigue. Large differences between the shape of power spectra of different muscles in the unfatigued state were found. These, however, were unrelated to endurance and degree of spectral shift during fatigue.


Reduction by copper supplementation of teratogenic effects of D-penicillamine.
Previous experiments from this laboratory demonstrated that oral administration of D-penicillamine (DPA) throughout gestation produced significant teratogenic effects and low copper concentrations in both maternal and fetal tissues in a dose-related manner. To test the hypothesis that the teratogenicity of DPA was due to induced copper deficiency, Sprague-Dawley rats were fed throughout gestation complete, purified diets containing 5 (control), 50, or 100 micrograms/g (supplemental) copper. One-half of the rats in each group received DPA at 0.83% of the diet. On day 21 of gestation, fetuses were removed and examined for gross malformations. Selected maternal and fetal tissues were analyzed for copper, zinc, iron and manganese. Animals fed the drug and supplemental copper had a very low incidence of fetal resorption (4%) and malformation (4%) compared to a high frequency of resorption (23%) and malformation (21%) in drug-fed nonsupplemented animals. There were no consistent differences in tissue iron, zinc or manganese levels among the groups. Tissue copper concentrations were lowest in the drug-fed, nonsupplemented group; those of the group fed DPA and supplemental copper were higher, but did not reach control values. Thus, there was a correlation between tissue copper levels and the frequency of malformations, supporting our hypothesis that DPA teratogenicity is due at least in part to induced copper deficiency.


Breast Cancer Risk Factors among Ugandan Women at a Tertiary Hospital: A Case-Control Study.
Although East Africa, like other countries in sub-Saharan Africa, has a lower incidence of breast cancer than high-income countries, the disease rate is rising steeply in Africa; it has nearly tripled in the past few decades in Uganda. There is a paucity of studies that have examined the relation between reproductive factors and breast cancer risk factors in Ugandan women. To determine breast cancer risk factors among indigenous Ugandan women. This is a hospital-based unmatched case-control study. Interviews were conducted between 2011 and 2012 using structured questionnaires. Patients with histology-proven breast cancer were recruited over a 2-year period. Logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs). A total of 350 women were recruited; 113 were cases and 237 were controls. The mean age was 47.5 years (SD 14) for the cases and 45.5 years (SD 14.1) for the controls. The odds of breast cancer risk seemed lower for those who breastfed (adjusted OR = 0.04; 95% CI: 0.01, 0.18). There was no significance for early age at first full-term birth (adjusted OR = 1.96; 95% CI: 0.97, 3.96; p = 0.061), and urban residence carried no increased odds of breast cancer either (p = 0.201). Breastfeeding seems to be associated with reduced odds of breast cancer.


When does less yield more? The impact of severity upon implicit recognition in pure alexia.
Pure alexia (PA) is characterised by strong effects of word length on reading times and is sometimes accompanied by an overt letter-by-letter (LBL) reading strategy. Past studies have reported "implicit recognition" in some individual PA patients. This is a striking finding because such patients are able to perform semantic classification and lexical decision at above chance levels even when the exposure duration is short enough to prevent explicit identification. In an attempt to determine the prevalence of this "implicit recognition" effect, we assessed semantic categorisation and lexical decision performance using limited exposure durations in 10 PA cases. The majority of the patients showed above chance accuracy in semantic categorisation and lexical decision. Performance on the lexical decision test was influenced by frequency and imageability. In addition, we found that the extent to which patients showed evidence of "implicit recognition" in both tasks was inversely related to the severity of their reading disorder. This result is consistent with hypotheses which suggest that this effect does not constitute an implicit form of unique word identification but is a reflection of the degree of partial activation within the word recognition system. These results also go some way towards explaining the individual variation in the presence of this effect observed across previous case-study investigations in the literature.


Results of total correction of the tetralogy of Fallot. Long-term haemodynamic evaluation at rest and during exercise.
Postoperative haemodynamic evaluation was performed on 15 patients 7 to 14 years (mean, 11 years) after total intracardiac repair of Fallot's tetralogy. The average age of the patients at the time of postoperative catheterization was 20 years (range 12 to 29) and they were all asymptomatic. Before operation 11 patients were severely cyanosed and disabled and 4 had mild cyanosis with anoxic spells. The cardiac output and right heart pressures were measured at rest and after 5 minutes of submaximal exercise on a bicycle ergometer. It was shown that the degree of residual obstruction of the pulmonary outflow tract was slight and that the right ventricular/pulmonary arterial systolic gradient (RV/PA gradient) exceeded 20 mmHg (2-6 kPa) at rest in only 3 patients. The mean cardiac index at rest was 5-4 l min-1 m-2 which increased to 8-3 l min-1 m-2 during exercise. The response of the cardiac output during exercise was either normal or supranormal in 14 of the 15 patients. In the remaining patient the response was marginally below the normal range. Pulmonary valvular regurgitation was present in 8 patients but there was no detectable difference in the cardiac function of these patients compared with patients with competent pulmonary valves. Six of the patients investigated had also had cardiac catheterizations performed 1 to 4 years after operation. The results of the present study show that remarkably little change has occurred in their haemodynamic status in the intervening years. Though these results are encouraging, further studies are required to determine the eventual outcome of the RV pressure and volume overload.


The effect of Mg2+ on cardiac muscle function: Is CaATP the substrate for priming myofibril cross-bridge formation and Ca2+ reuptake by the sarcoplasmic reticulum?
Kinetics are established for the activation of the myofibril from the relaxed state [Smith, Dixon, Kirschenlohr, Grace, Metcalfe and Vandenberg (2000) Biochem. J. 346, 393-402]. These require two troponin Ca2+-binding sites, one for each myosin head, to act as a single unit in initial cross-bridge formation. This defines the first, or activating, ATPase reaction, as distinct from the further activity of the enzyme that continues when a cross-bridge to actin is already established. The pairing of myosin heads to act as one unit suggests a possible alternating mechanism for muscle action. A large positive inotropic (contraction-intensifying) effect of loading the Mg2+ chelator citrate, via its acetoxymethyl ester, into the heart has confirmed the competitive inhibition of the Ca2+ activation by Mg2+, previously seen in vitro. In the absence of a recognized second Ca2+ binding site on the myofibril, with appropriate binding properties, the bound ATP is proposed as the second activating Ca2+-binding site. As ATP, free or bound to protein, can bind either Mg2+ or Ca2+, this leads to competitive inhibition by Mg2+. Published physico-chemical studies on skeletal muscle have shown that CaATP is potentially a more effective substrate than MgATP for cross-bridge formation. The above considerations allow calculation of the observed variation of fractional activation by Ca2+ as a function of [Mg2+] and in turn reveal simple Michaelis-Menten kinetics for the activation of the ATPase by sub-millimolar [Mg2+]. Furthermore the ability of bound ATP to bind either cation, and the much better promotion of cross-bridge formation by CaATP binding, give rise to the observed variation of the Hill coefficient for Ca2+ activation with altered [Mg2+]. The inclusion of CaADP within the initiating cross-bridge and replacement by MgADP during the second cycle is consistent with the observed fall in the rate of the myofibril ATPase that occurs after two phosphates are released. The similarity of the kinetics of the cardiac sarcoplasmic reticulum ATPase to those of the myofibril, in particular the positive co-operativity of both Mg2+ inhibition and Ca2+ activation, leads to the conclusion that this ATPase also has an initiation step that utilizes CaATP. The first-order activation by sub-millimolar [Mg2+], similar to that of the myofibril, may be explained by Mg2+ involvement in the phosphate-release step of the ATPase. The inhibition of both the myofibril and sarcoplasmic reticulum Ca2+ transporting ATPases by Mg2+ offers an explanation for the specific requirement for phosphocreatine (PCr) for full activity of both enzymes in situ and its effect on their apparent affinities for ATP. This explanation is based on the slow diffusion of Mg2+ within the myofibril and on the contrast of PCr with both ATP and phosphoenolpyruvate, in that PCr does not bind Mg2+ under physiological conditions, whereas both the other two bind it more tightly than the products of their hydrolysis do. The switch to supply of energy by diffusion of MgATP into the myofibril when depletion of PCr raises [ATP]/[PCr] greatly, e.g. during anoxia, results in a local [Mg2+] increase, which inhibits the ATPase. It is possible that mechanisms similar to those described above occur in skeletal muscle but the Ca2+ co-operativity involved would be masked by the presence of two Ca2+ binding sites on each troponin.


Actin-associated neurabin-protein phosphatase-1 complex regulates hippocampal plasticity.
Protein phosphatase-1 (PP1) has been implicated in the control of long-term potentiation (LTP) and depression (LTD) in rat hippocampal CA1 neurons. PP1 catalytic subunits associate with multiple postsynaptic regulatory subunits, but the PP1 complexes that control hippocampal LTP and LTD in the rat hippocampus remain unidentified. The neuron-specific actin-binding protein, neurabin-I, is enriched in dendritic spines, and tethers PP1 to actin-rich postsynaptic density to regulate morphology and maturation of spines. The present studies utilized Sindbis virus-mediated expression of wild-type and mutant neurabin-I polypeptides in organotypic cultures of rat hippocampal slices to investigate their role in synaptic plasticity. While wild-type neurabin-I elicited no change in basal synaptic transmission, it enhanced LTD and inhibited LTP in CA1 pyramidal neurons. By comparison, mutant neurabins, specifically those unable to bind PP1 or F-actin, decreased basal synaptic transmission, attenuated LTD and increased LTP in slice cultures. Biochemical and cell biological analyses suggested that, by mislocalizing synaptic PP1, the mutant neurabins impaired the functions of endogenous neurabin-PP1 complexes and modulated LTP and LTD. Together, these studies provided the first biochemical and physiological evidence that a postsynaptic actin-bound neurabin-I-PP1 complex regulates synaptic transmission and bidirectional changes in hippocampal plasticity.


The relevance of the Heart Foundation of Australia's dietary recommendations for adult Australians: a comparison of views of general practitioners, cardiologists and dietitians.
To compare the views of general practitioners, cardiologists and dietitians about the relevance of the Heart Foundation of Australia's dietary recommendations for adult cardiac patients. Quantitative-cross sectional study. Postal questionnaires were self-completed by 248 Victorian general practitioners (30% response), 189 Australia-wide cardiologists (47% response) and 180 Victorian dietitians (45% response). Responses were represented as percentages and analyses of variance were conducted to explore the impact of the independent variables: age, work status and gender on the dependent variable: dietary recommendation. Approximately half of the recommendations were viewed as strongly important to implement; these related to lean meats, limiting takeaways and cakes/biscuits, and adjusting energy intake. Others of importance were eating fruits, vegetables and fish. However, most of these goals were seen as difficult to achieve. Dietitians appeared to share responses of doctors, except for greater importance of eating fruit and vegetables and a greater difficulty in limiting cakes and biscuits. There was a high level of agreement among the three groups (mean 87%) about patients having difficulty implementing adjusting energy intake. There is agreement amongst these professionals that many of the recommendations lack importance, specifically those pertaining to unsaturated oils, low fat dairy products, cholesterol rich foods, intake of legumes and grains and the restriction of salt. This may reflect a need for further nutrition education.


Interaction of dietary saturated and polyunsaturated triglycerides in regulating the processes that determine plasma low density lipoprotein concentrations in the rat.
These studies were undertaken to determine how polyunsaturated (n-3 and n-6) and saturated triglycerides interact to regulate rates of low density lipoprotein (LDL) production and rates of receptor-dependent and receptor-independent LDL transport. Animals were fed diets containing 20% (by wt) hydrogenated coconut oil or diets in which the coconut oil was progressively removed and replaced with safflower oil or fish oil concentrate. Plasma LDL concentrations fell when either of the polyunsaturated triglycerides was substituted for saturated triglycleride in the diet; however, the reduction in LDL concentrations was greater with fish oil than with safflower oil at all ratios of polyunsaturated to saturated triglyceride that were examined. The lower plasma LDL concentrations when coconut oil was replaced with fish oil could be attributed almost entirely to a much greater increase in hepatic LDL receptor activity when fish oil was used as the substitute than when safflower oil was used as the substitute. To examine the effect of polyunsaturated triglycerides when used to supplement a high saturated fat diet rather than to replace saturated fat in the diet, animals were fed a diet containing 15% coconut oil (by wt) with or without an additional supplement of 5% fish oil or safflower oil. The addition of 15% coconut oil to low fat control diet increased the rate of LDL production causing circulating LDL levels to rise by 40%. The further supplementation of this high saturated fat diet with fish oil concentrate markedly increased hepatic LDL receptor activity causing plasma LDL concentrations to return to control values whereas supplementation with safflower oil had little effect. Thus, at least in the rat, supplementation of a high saturated fat diet with a fish oil concentrate lowers plasma LDL concentrations as effectively as removing the saturated fat from the diet, although in the former case, both the production and the receptor-dependent uptake of LDL are greatly increased.


Rapid dynamic image registration of the beating heart for diagnosis and surgical navigation.
Dynamic cardiac magnetic resonance imaging (MR) and computed tomography (CT) provide cardiologists and cardiac surgeons with high-quality 4-D images for diagnosis and therapy, yet the effective use of these high-quality anatomical models remains a challenge. Ultrasound (US) is a flexible imaging tool, but the US images produced are often difficult to interpret unless they are placed within their proper 3-D anatomical context. The ability to correlate real-time 3-D US volumes (RT3D US) with dynamic MR/CT images would offer a significant contribution to improve the quality of cardiac procedures. In this paper, we present a rapid two-step method for registering RT3D US to high-quality dynamic 3-D MR/CT images of the beating heart. This technique overcomes some major limitations of image registration (such as the correct registration result not necessarily occurring at the maximum of the mutual information (MI) metric) using the MI metric. We demonstrate the effectiveness of our method in a dynamic heart phantom (DHP) study and a human subject study. The achieved mean target registration error of CT+US images in the phantom study is 2.59 mm. Validation using human MR/US volumes shows a target registration error of 1.76 mm. We anticipate that this technique will substantially improve the quality of cardiac diagnosis and therapies.


Metabolic fate of cysteine and methionine in rumen digesta.
Estimates were obtained of the extent to which cysteine and methionine were incorporated into the protein of the microbes of rumen digesta without prior degradation and resynthesis. By using the amino acids labeled with both (35)S and (14)C, it was observed that a large proportion of the (35)S appeared in the sulfide pool and of the (14)C appeared in volatile fatty acids. By isolating the appropriate amino acid, obtaining the (14)C to (35)S ratio, and comparing this with the ratio in the added amino acid, the degree of direct incorporation was calculated. For cysteine it was estimated that at most 1% and for methionine, at most 11% of the amino acid in the free pool was incorporated unchanged into microbial protein. As a consequence of these findings, it is considered that the method for measuring microbial protein synthesis in rumen digesta based upon incorporation of (35)S from the free sulfide pool is not seriously affected by direct utilization of sulfur amino acids arising from dietary sources.


Multispectral infrared reflectography to differentiate features in paintings.
Infrared reflectography is a well-known technique based on wideband imaging in the near-infrared (NIR) range used for painting diagnostics in conservation laboratories.. This work is focused on the application of multiband reflectography for analysis of pictorial layers and differentiated detection of painting features. This technique generates a set of narrowband NIR images of the painting. Starting from a dataset that is registered, metrically correct, and calibrated, the capability of collecting both spectral and spatial information has been exploited by processing the image cube with interplane techniques. Examples on artworks by Caravaggio, Veronese, Bronzino, and Schiavone are presented.


Evidence for functional diversity between the voltage-gated proton channel Hv1 and its closest related protein HVRP1.
The Hv1 channel and voltage-sensitive phosphatases share with voltage-gated sodium, potassium, and calcium channels the ability to detect changes in membrane potential through voltage-sensing domains (VSDs). However, they lack the pore domain typical of these other channels. NaV, KV, and CaV proteins can be found in neurons and muscles, where they play important roles in electrical excitability. In contrast, VSD-containing proteins lacking a pore domain are found in non-excitable cells and are not involved in neuronal signaling. Here, we report the identification of HVRP1, a protein related to the Hv1 channel (from which the name Hv1 Related Protein 1 is derived), which we find to be expressed primarily in the central nervous system, and particularly in the cerebellum. Within the cerebellar tissue, HVRP1 is specifically expressed in granule neurons, as determined by in situ hybridization and immunohistochemistry. Analysis of subcellular distribution via electron microscopy and immunogold labeling reveals that the protein localizes on the post-synaptic side of contacts between glutamatergic mossy fibers and the granule cells. We also find that, despite the similarities in amino acid sequence and structural organization between Hv1 and HVRP1, the two proteins have distinct functional properties. The high conservation of HVRP1 in vertebrates and its cellular and subcellular localizations suggest an important function in the nervous system.


To Expand the Evidence Base About Harms from Tests and Treatments.
Rigorous evidence about the broad range of harms that might be experienced by a patient in the course of testing and treatment is sparse. We aimed to generate recommendations for how researchers might more comprehensively evaluate potential harms of healthcare interventions, to allow clinicians and patients to better include this evidence in clinical decision-making. We propose seven domains of harms of tests and treatments that are relevant to patients: (1) physical impairment, (2) psychological distress, (3) social disruption, (4) disruption in connection to healthcare, (5) labeling, (6) financial impact, and (7) treatment burden. These domains will include a range of severity of harms and variation in timing after testing or treatment, attributable to the service itself or a resulting care cascade. Although some new measures may be needed, diverse data and tools are available to allow the assessment of harms comprehensively across these domains. We encourage researchers to evaluate harms in sub-populations, since the harms experienced may differ importantly by demographics, social determinants, presence of comorbid illness, psychological state, and other characteristics. Regulators, funders, and editors might require either assessment or reporting of harms in each domain or require justification for inclusion and exclusion of different domains.


Selenium promotes metabolic conversion of T-2 toxin to HT-2 toxin in cultured human chondrocytes.
To explore the metabolism of T-2 toxin in human chondrocytes (HCs) and determine the impact of selenium supplementation. For determination of cytotoxicity using the MTT assay, optical density values were read with an automatic enzyme-linked immunosorbent assay reader at 510nm. Cell survival was calculated and the cytotoxicity estimated. To identify the metabolites of T-2 toxin, the medium supernatants and C28/I2 cells were analyzed by high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) separately. For HPLC-MS/MS, the mobile phase A was water and phase B was 98% methanol. The gradient for the elution was: 0-0.5min, 50% of B; 0.5-2.0min, 100% of B; 2.0-3.5min, 100% of B; 3.6-6min, 50% of B. T-2 toxin increased the toxicity to C28/I2 cells significantly in a dose- and time-dependent manner (viability range 91.5-22.0%). Supplementation with selenium (100ng/mL) could increase the cell viability after the 24h incubation. The concentration of T-2 toxin in the cell medium decreased from 20 to 6.67±1.02ng/mL, and the concentration of HT-2 toxin increased from 0 to 6.88±1.23ng/mL during the 48h incubation, whereas the relative concentration of T-2 toxin in cells increased from 0 to 12.80±1.84ng/g. Supplementary selenium in the HCs cultures reduced the cytotoxicity induced by T-2 toxin significantly, and was associated with rapid conversion of T-2 toxin in the culture medium to HT-2 toxin. T-2 toxin was more toxic to HCs than HT-2 toxin at equivalent concentrations. HT-2 toxin was a detectable metabolite of T-2 toxin in cultured HCs, and selenium enhanced the metabolic conversion of T-2 toxin, reducing its cytotoxicity to HCs.


Release of tumor necrosis factor-alpha from macrophages. Enhancement and suppression are dose-dependently regulated by prostaglandin E2 and cyclic nucleotides.
PGE2 has previously been shown to suppress various leukocyte functions. In this study, we examined whether PGE2 would affect release of TNF-alpha from rat resident peritoneal macrophages. Two different, dose-dependent effects were observed: low PGE2 concentrations (0.1 to 10 ng/ml) stimulated, whereas higher concentrations (greater than 10 ng/ml) suppressed TNF-alpha release. PGE2-stimulated TNF-alpha production was dependent on de novo protein synthesis and was associated with an intracellular rise of cGMP. The importance of cGMP as an intracellular messenger for PGE2 was confirmed by the following evidence: (1) low PGE2 concentrations preferentially increased cGMP and not cAMP and (2) cGMP, either exogenously added or endogenously generated by sodium nitroprusside, were efficient stimulators of TNF-alpha production. In contrast, agents increasing intracellular cAMP concentrations such as PGE1, higher PGE2 doses, isoproterenol, and theophylline, all suppressed TNF-alpha synthesis. Only resident, but not casein-elicited or Corynebacterium parvum-activated macrophages, were stimulated by low PGE2 concentrations to increase TNF-alpha production. In tumor cytotoxicity assays, PGE2-activated macrophages were active only against TNF-alpha-sensitive target cells. These findings demonstrate that TNF-alpha synthesis in macrophages is up-regulated by cGMP and down-regulated by cAMP, which indicates that cyclic nucleotides act as intracellular messengers for extracellular signals of macrophage activation.


Surgical approaches to aortopexy for severe tracheomalacia.
The purpose of this study was to determine the outcomes among three different surgical approaches for performing an aortopexy to treat severe tracheomalacia (STM). A retrospective review was performed for all patients who underwent an aortopexy by pediatric surgeons at a single institution during 1997-2012. Data collected included details of the operative approaches and clinical results. The data were analyzed using Chi-square and Fisher exact test. Forty-one patients underwent an aortopexy. The operation was chosen by the surgeon and not randomized. Exposure was by partial sternotomy (PS) (20), open thoracotomy (12), or thoracoscopic approach (7). Only the PS approach was done by a single team. All groups showed improvement in work of breathing, prevention of severe respiratory distress, and acute life threatening events. These effects were more dramatic for the PS group, especially regarding oxygen and/or ventilator dependence and the ability to undergo tracheostomy decannulation. Among the sixteen patients with failure-to-thrive before successful aortopexy by any technique, ten demonstrated significant improvement in their growth (p=0.025). The recurrence rate for the thoracoscopic approach was 38%, and there were no recurrences in the partial sternotomy and the thoracotomy groups, 38% vs 0% vs 0%, p=0.005. Simultaneous bronchoscopy was utilized more commonly in the PS group compared to the thoracotomy and thoracoscopic group, 95% vs 62% vs 38%. In this series, the partial sternotomy technique had the most reliable resolution of symptoms and no recurrence requiring reoperation. The PS approach to STM has the technical advantages of an improved exposure with equal access to the vessels over the right and left mainstem bronchi, as well as the trachea and a more specific elevation of the arteries, including suspension of the pulmonary arteries and trachea itself when desirable. Simultaneous bronchoscopy during aortopexy and an experienced team also likely contribute to improved outcomes. The variations in populations, follow-up, and use of continuous intraoperative bronchoscopy, however, make firm conclusions difficult.


Electroreductive hydroxy fluorosulfonylation of alkenes.
An electroreductive strategy for radical hydroxyl fluorosulfonylation of alkenes with sulfuryl chlorofluoride and molecular oxygen from air is described. This mild protocol displays excellent functional group compatibility, broad scope, and good scalability, providing convenient access to diverse β-hydroxy sulfonyl fluorides. These β-hydroxy sulfonyl fluoride products can be further converted to valuable aliphatic sulfonyl fluorides, β-keto sulfonyl fluorides, and β-alkenyl sulfonyl fluorides. Further, some of these products showed excellent inhibitory activity against Botrytis cinerea or Bursaphelenchus xylophilus, which could be useful for potent agrochemical discovery. Preliminary mechanistic studies indicate that this transformation is achieved through rapid O2 interception by the alkyl radical and subsequent reduction of the peroxy radical, which outcompete other side reactions such as chlorine atom transfer, hydrogen atom transfer, and Russell fragmentation.


Mesenchymal stem cell application in children with subacute sclerosing panencephalitis.
Subacute sclerosing panencephalitis (SSPE) is a serious, often fatal disease that responds poorly to current treatment modalities. Recently, the ability of mesenchymal stem cells (MSCs) to produce neurotrophic factors and inflammatory molecules has placed them among potential treatment agents for neurological conditions. We report the results of four patients treated with MSC for SSPE. The patients were followed up clinically, and by periodical laboratory evaluations, magnetic resonance imaging (MRI), and electroencephalography. One patient deteriorated to stage III of the disease, two patients remained in the same stage, and one died from disease progression and respiratory problems. Neurological findings and electroencephalography scores were consistent with the clinical course of the patient whereas MRI showed new inflammatory lesions in two patients. This is the first report of the application of MSC in SSPE. No benefit is demonstrated.


Chemical basis of pupal cannibalism in a caterpillar (Utetheisa ornatrix).
The moth Utetheisa ornatrix derives protection against predation from systemic pyrrolizidine alkaloids (PAs) that it sequesters as a larva from its foodplants (Leguminosae, Crotalaria spp.). We here show, in laboratory tests, that Utetheisa deficient in body PA can make up for the chemical shortfall by cannibalizing pupae. We present evidence indicating that cannibalism in larvae is elicited not by hunger, but possibly by PA deficiency itself, and that in making cannibalistic choices larvae prefer PA-containing over PA-free pupae. PAs themselves, either in crystalline form or as additives to food items, proved phagostimulatory to larvae. In nature Utetheisa tend to pupate away from their foodplant, essentially out of reach of larval attack. The threat of cannibalism may have contributed to the evolution of this pupation behavior.


Comparison of microsuture, interpositional nerve graft, and laser solder weld repair of the rat inferior alveolar nerve.
Intraosseous repair of nerves involves difficulty of access and there is concern that bone healing may interfere with repair outcomes. The present report describes the effect of 3 separate repair techniques on recovery from section of the rat intraosseous inferior alveolar nerve, with reference to the mental nerve distal and the trigeminal ganglion proximal to the nerve section. Unilateral exposure of the inferior alveolar nerves of 28 rats was achieved through bone windows. Nerves were sectioned and rats were assigned to 1 of 4 groups (n = 7): untreated controls, microsuture repair, interpositional nerve grafts from the femoral nerve, or laser solder weld repair. Animals were sacrificed 1 year after surgery for histologic evaluation of the mental nerve, inferior alveolar nerve, and trigeminal ganglion compared with unoperated contralateral nerves. Compared with the unoperated contralateral nerves, nerve section substantially decreased mental nerve fiber number, mental nerve myelination, mental nerve fiber diameter, inferior alveolar nerve vascularity, trigeminal neuron number, and trigeminal neuron horseradish peroxidase tracer uptake and increased trigeminal ganglion degenerate neurons (P < .001). All 3 forms of repair substantially decreased these effects (P < .05). Interpositional nerve graft was least effective (P < .05). Nonetheless, mental nerve fiber diameter was significantly decreased compared with unsectioned nerves after microsuture and laser solder weld repair (P < .05). Intraosseous repair of the inferior alveolar nerve decreases peripheral and central signs of degeneration. Clinical hyperesthesia after repair may reflect a predominance of small fibers after recovery.


Effects of HIV infection on the metabolic and hormonal status of children with severe acute malnutrition.
HIV infection occurs in 30% of children with severe acute malnutrition in sub-Saharan Africa. Effects of HIV on the pathophysiology and recovery from malnutrition are poorly understood. We conducted a prospective cohort study of 75 severely malnourished Ugandan children. HIV status/CD4 counts were assessed at baseline; auxologic data and blood samples were obtained at admission and after 14 days of inpatient treatment. We utilized metabolomic profiling to characterize effects of HIV infection on metabolic status and subsequent responses to nutritional therapy. At admission, patients (mean age 16.3 mo) had growth failure (mean W/H z-score -4.27 in non-edematous patients) that improved with formula feeding (mean increase 1.00). 24% (18/75) were HIV-infected. Nine children died within the first 14 days of hospitalization; mortality was higher for HIV-infected patients (33% v. 5%, OR = 8.83). HIV-infected and HIV-negative children presented with elevated NEFA, ketones, and even-numbered acylcarnitines and reductions in albumin and amino acids. Leptin, adiponectin, insulin, and IGF-1 levels were low while growth hormone, cortisol, and ghrelin levels were high. At baseline, HIV-infected patients had higher triglycerides, ketones, and even-chain acylcarnitines and lower leptin and adiponectin levels than HIV-negative patients. Leptin levels rose in all patients following nutritional intervention, but adiponectin levels remained depressed in HIV-infected children. Baseline hypoleptinemia and hypoadiponectinemia were associated with increased mortality. Our findings suggest a critical interplay between HIV infection and adipose tissue storage and function in the adaptation to malnutrition. Hypoleptinemia and hypoadiponectinemia may contribute to high mortality rates among malnourished, HIV-infected children.


Tests of adrenal insufficiency.
In suspected adrenal insufficiency, the ideal test for assessing the hypothalamo-pituitary-adrenal axis is controversial. Therefore, three tests were compared in patients presenting with symptoms suggestive of adrenal insufficiency. Responses to the standard short Synacthen test (SSST), the low dose Synacthen test (LDST), and the 08:00 hour serum cortisol concentration were measured in 32 patients. A normal response to the synacthen test was defined as a peak serum cortisol of >/= 500 nmol/l and/or incremental concentration of >/= 200 nmol/l. The sensitivity and specificity of the 08:00 hour serum cortisol concentration compared with other tests was calculated. Three patients had neither an adequate peak nor increment after the SSST and LDST. All had a serum 08:00 hour cortisol concentration of < 200 nmol/l. Eight patients had abnormal responses by both criteria to the LDST but had normal responses to the SSST. Three reported amelioration of their symptoms on hydrocortisone replacement. Twenty one patients had a normal response to both tests (of these, 14 achieved adequate peak and increment after both tests and seven did not have an adequate peak after the LDST but had a normal increment). The lowest 08:00 hour serum cortisol concentration above which patients achieved normal responses to both the LDST and SSST was 500 nmol/l. At this cut off value (compared with the LDST), the serum 08:00 hour cortisol concentration had a sensitivity of 100% but specificity was only 33%. The LDST revealed mild degrees of adrenal insufficiency not detected by the SSST. The value of a single 08:00 hour serum cortisol concentration is limited.


Mycobacterium marinum escapes from phagosomes and is propelled by actin-based motility.
Mycobacteria are responsible for a number of human and animal diseases and are classical intracellular pathogens, living inside macrophages rather than as free-living organisms during infection. Numerous intracellular pathogens, including Listeria monocytogenes, Shigella flexneri, and Rickettsia rickettsii, exploit the host cytoskeleton by using actin-based motility for cell to cell spread during infection. Here we show that Mycobacterium marinum, a natural pathogen of fish and frogs and an occasional pathogen of humans, is capable of actively inducing actin polymerization within macrophages. M. marinum that polymerized actin were free in the cytoplasm and propelled by actin-based motility into adjacent cells. Immunofluorescence demonstrated the presence of host cytoskeletal proteins, including the Arp2/3 complex and vasodilator-stimulated phosphoprotein, throughout the actin tails. In contrast, Wiskott-Aldrich syndrome protein localized exclusively at the actin-polymerizing pole of M. marinum. These findings show that M. marinum can escape into the cytoplasm of infected macrophages, where it can recruit host cell cytoskeletal factors to induce actin polymerization leading to direct cell to cell spread.


The dynamic range of human lightness perception.
Natural viewing challenges the visual system with images that have a dynamic range of light intensity (luminance) that can approach 1,000,000:1 and that often exceeds 10,000:1 [1, 2]. The range of perceived surface reflectance (lightness), however, can be well approximated by the Munsell matte neutral scale (N 2.0/ to N 9.5/), consisting of surfaces whose reflectance varies by about 30:1. Thus, the visual system must map a large range of surface luminance onto a much smaller range of surface lightness. We measured this mapping in images with a dynamic range close to that of natural images. We studied simple images that lacked segmentation cues that would indicate multiple regions of illumination. We found a remarkable degree of compression: at a single image location, a stimulus luminance range of 5,905:1 can be mapped onto an extended lightness scale that has a reflectance range of 100:1. We characterized how the luminance-to-lightness mapping changes with stimulus context. Our data rule out theories that predict perceived lightness from luminance ratios or Weber contrast. A mechanistic model connects our data to theories of adaptation and provides insight about how the underlying visual response varies with context.


BLM interaction with EZH2 regulates MDM2 expression and is a poor prognostic biomarker for prostate cancer.
Prostate cancer (PCa) is one of the major causes of cancer death among males worldwide. Our previous studies indicated that the proliferation of prostate cancer cells was reduced after BLM knockdown, however, the mechanism is still not clear. In this study, we identified a direct interaction between BLM and EZH2, which had extremely significantly positive correlations (P<0.001). In vitro, our research revealed that tumor growth was inhibited after EZH2 knockdown and that inhibition could be reversed by BLM overexpression; conversely, tumor growth was promoted after EZH2 overexpression, and promotion could be reversed by BLM knockdown. This suggests that BLM and EZH2 play important roles in the progression of prostate cancer cells. In vivo, the impact of BLM and EZH2 was investigated in mouse xenograft models, and the results showed that EZH2 could be regulated by BLM, which was consistent with our in vitro observations. Our results demonstrated that the expression of P53 is affected by the binding of BLM and EZH2 to the MDM2 promoter region. This finding indicated that EZH2 regulates the expression of MDM2 at the transcriptional level by interacting with BLM.


Detecting optic disc on asians by multiscale gaussian filtering.
The optic disc (OD) is an important anatomical feature in retinal images, and its detection is vital for developing automated screening programs. Currently, there is no algorithm designed to automatically detect the OD in fundus images captured from Asians which are larger and have thicker vessels compared to Caucasians. In this paper, we propose such a method to complement current algorithms using two steps: OD vessel candidate detection and OD vessel candidate matching. The first step is achieved with multiscale Gaussian filtering, scale production, and double thresholding to initially extract the vessels' directional map of various thicknesses. The map is then thinned before another threshold is applied to remove pixels with low intensities. This result forms the OD vessel candidates. In the second step, a Vessels' Directional Matched Filter (VDMF) of various dimensions is applied to the candidates to be matched, and the pixel with the smallest difference designated the OD center. We tested the proposed method on a new database consisting of 402 images from a diabetic retinopathy (DR) screening programme consisting of Asians. The OD center was successfully detected with an accuracy of 99.25% (399/402).


The value of magnetic resonance imaging and ultrasound in undifferentiated arthritis: a systematic review.
To perform a systematic literature review of the diagnostic and prognostic value of magnetic resonance imaging (MRI) and ultrasound (US) in patients with undifferentiated peripheral inflammatory arthritis (UPIA), and to assess if MRI and US should be done at baseline and repeated, and if so, at what interval. Medline, Embase, the Cochrane Library, and abstracts presented at the 2007 and 2008 meetings of the American College of Rheumatology and European League Against Rheumatism meetings were searched for diagnostic and prognostic studies of any duration examining the ability of MRI/US to predict outcome of patients with UPIA. Sensitivity, specificity, predictive values, and positive/negative likelihood ratios (LR+/LR-) were calculated. When available, odds ratios were extracted. Quality was appraised using validated scales. Regarding MRI, 11 out of 2595 screened references were included: 2 described pure undifferentiated arthritis (UA) populations and 9, mixed populations. Bone edema (LR+ 4.5) and combination of a distinct MRI synovitis and erosion pattern (LR+ 4.8) increased probability of developing rheumatoid arthritis (RA). Absence of MRI synovitis (LR- 0.2) and absence of a distinct synovitis pattern (LR- 0) decreased probability of developing RA. Regarding US, 2 out of 2111 references were included, both mixed populations; no data could be extrapolated for UPIA. MRI bone edema and combined synovitis and erosion pattern seem useful in predicting development of RA from UPIA. The value of US in UPIA remains to be determined. The absence of MRI synovitis seems useful in excluding development of RA. No data were found about the value of repeating MRI/US. Studies evaluating MRI/US in UPIA are scarce, but current knowledge strongly encourages further testing in UA.


Dual therapy with zinc acetate and rifaximin prevents from ethanol-induced liver fibrosis by maintaining intestinal barrier integrity.
Hepatic overload of gut-derived lipopolysaccharide dictates the progression of alcoholic liver disease (ALD) by inducing oxidative stress and activating Kupffer cells and hepatic stellate cells through toll-like receptor 4 signaling. Therefore, targeting the maintenance of intestinal barrier integrity has attracted attention for the treatment of ALD. Zinc acetate and rifaximin, which is a nonabsorbable antibiotic, had been clinically used for patients with cirrhosis, particularly those with hepatic encephalopathy, and had been known to improve intestinal barrier dysfunction. However, only few studies focused on their efficacies in preventing the ALD-related fibrosis development. To investigate the effects of a combined zinc acetate with rifaximin on liver fibrosis in a mouse ALD model. To induce ALD-related liver fibrosis, female C57BL/6J mice were fed a 2.5% (v/v) ethanol-containing Lieber-DeCarli liquid diet and received intraperitoneal carbon tetrachloride (CCl4) injection twice weekly (1 mL/kg) for 8 wk. Zinc acetate (100 mg/L) and/or rifaximin (100 mg/L) were orally administered during experimental period. Hepatic steatosis, inflammation and fibrosis as well as intestinal barrier function were evaluated by histological and molecular analyses. Moreover, the direct effects of both agents on Caco-2 barrier function were assessed by in vitro assays. In the ethanol plus CCl4-treated mice, combination of zinc acetate and rifaximin attenuated oxidative lipid peroxidation with downregulation of Nox2 and Nox4. This combination significantly inhibited the Kupffer cells expansion and the proinflammatory response with blunted hepatic exposure of lipopolysaccharide and the toll-like receptor 4/nuclear factor kB pathway. Consequently, liver fibrosis and hepatic stellate cells activation were efficiently suppressed with downregulation of Mmp-2, -9, -13, and Timp1. Both agents improved the atrophic changes and permeability in the ileum, with restoration of tight junction proteins (TJPs) by decreasing the expressions of tumor necrosis factor α and myosin light chain kinase. In the in vitro assay, both agents directly reinforced ethanol or lipopolysaccharide-stimulated paracellular permeability and upregulated TJPs in Caco-2 cells. Dual therapy with zinc acetate and rifaximin may serve as a strategy to prevent ALD-related fibrosis by maintaining intestinal barrier integrity.


Cost containment of the second-generation cephalosporins by prospective monitoring at a community teaching hospital.
All patients receiving cefoxitin and cefamandole were prospectively reviewed for appropriate and inappropriate utilization. There were two eight-week survey periods. In period 1, 81 (70%) of 115 patients received cefoxitin appropriately and six (40%) of 15 patients received cefamandole appropriately. In patients receiving antibiotics inappropriately, 12 (35%) of the 34 receiving cefoxitin and eight (89%) of the nine receiving cefamandole had infections that could have been treated with less expensive, equally efficacious antibiotics. Changes in antibiotic therapy were made in 79% of patients based on our recommendations. The estimated annual cost saving for these antibiotics was $40,290. During period 2, 73 (91%) of 80 patients were given cefoxitin appropriately and 14 (61%) of 23 patients received cefamandole appropriately. Forty-three percent of those receiving cefoxitin and 33% of those receiving cefamandole inappropriately could have been treated with a less expensive, equally efficacious antibiotic. In 88% of patients, the attending physicians followed our recommendations.


Structural relatedness between the 18S rRNA genes and the formyl peptide receptor genes: new insights into the phylogenesis of immune receptors.
In this study the authors examined the sequences of the ribosomal 18S rRNA of Drosophila and man and 16 mRNA sequences coding for different members of the family of the mammalian formyl peptide receptors (FPRs). The positions in the sequences of all >or=7-base oligonucleotide identities occurring in at least one of the 18S rRNAs and one of the FPR mRNAs were recorded. On the basis of the positional data, the Drosophila 18S-FPR and human 18S-FPR distances (in nucleotides) were determined for each identity. Then the actual frequency distribution of the distances (grouped into 200-unit classes) was derived. The theoretical frequency distribution of distances was also calculated under the assumption of non-relatedness between the 18S and FPR sequences. Comparison between the theoretical and the actual distributions showed that at class -500 (range from - 400 to - 600) of the 18S-FPR values the actual frequency was significantly (p < 0.01) higher than the theoretical frequency, in both Drosophila and man, suggesting that the second section of the FPR genes (approximately from nucleotide 400 to the end of sequence) may be structurally related to the first section of the ribosomal 18S genes (approximately nucelotides 1-650). The authors advance the hypothesis that the two families of genes may have used common ancestral raw genetic materials in the building of the extant sequences.


Ubiquitous fiber exposure in selected sampling sites in Europe.
This study evaluates personal exposure to respirable inorganic and organic fibers during normal human lifetimes and assesses the order of magnitude of the contribution of inorganic fibers other than asbestos to total fiber exposure from man-made and natural sources. Four groups (suburban schoolchildren, rural retired persons, office workers, and taxi drivers), with five persons per group, were monitored for 24 h four times during one year. Personal sampling pumps collected airborne dust on gold-precoated Nuclepore filters. The fibers were analyzed for fiber sizes specified by the World Health Organization. The geometric mean concentrations ranged from 9000 fibers.m-3 (office workers) to 20000 fibers.m-3 (schoolchildren) for organic fibers, and from 600 fibers.m-3 (taxi drivers) to 4000 fibers.m-3 (schoolchildren) for gypsum fibers. For other inorganic fibers the concentrations were around 5000 fibers.m-3. The contribution of fibers with an elemental composition similar to that of man-made vitreous fibers (MMVF) was less than about one-quarter of the content of other inorganic fibers. The fiber size distributions were uniform across the groups, and the organic fibers were the longest and thinnest nonasbestos fibers. Lifetime exposure to fibers can be ranked as organic fibers > other inorganic fibers > fibers with an elemental composition similar to MMVF > MMVF. Information on the biological effects of fibers is difficult to interpret for use in assessing the health risk from exposure to low levels of ubiquitous fibers, and there is a lack of knowledge on the effects of organic fibers.


Chikungunya as a cause of acute febrile illness in southern Sri Lanka.
Chikungunya virus (CHIKV) re-emerged in Sri Lanka in late 2006 after a 40-year hiatus. We sought to identify and characterize acute chikungunya infection (CHIK) in patients presenting with acute undifferentiated febrile illness in unstudied rural and semi-urban southern Sri Lanka in 2007. We enrolled febrile patients ≥ 2 years of age, collected uniform epidemiologic and clinical data, and obtained serum samples for serology, virus isolation, and real-time reverse-transcriptase PCR (RT-PCR). Serology on paired acute and convalescent samples identified acute chikungunya infection in 3.5% (28/797) patients without acute dengue virus (DENV) infection, 64.3% (18/28) of which were confirmed by viral isolation and/or real-time RT-PCR. No CHIKV/DENV co-infections were detected among 54 patients with confirmed acute DENV. Sequencing of the E1 coding region of six temporally distinct CHIKV isolates (April through October 2007) showed that all isolates posessed the E1-226A residue and were most closely related to Sri Lankan and Indian isolates from the same time period. Except for more frequent and persistent musculoskeletal symptoms, acute chikungunya infections mimicked DENV and other acute febrile illnesses. Only 12/797 (1.5%) patients had serological evidence of past chikungunya infection. Our findings suggest CHIKV is a prominent cause of non-specific acute febrile illness in southern Sri Lanka.


Simple procedure for production by group C streptococci of phage-associated lysin active against group A streptococci.
Phage-associated lysin of high potency was prepared by growing the host group C streptococcal strain 26RP66 in a semisynthetic medium. The lysin was stabilized by adding dithiothreitol and neutralized ethylenediaminetetraacetic acid (EDTA) to facilitate further concentration and partial purification. The lysin remained active when stored at -65 C for 1 year. Lysin was active against all strains of group A streptococci tested and was more active against living cells than heat-killed cells. The procedure outlined is practicable for most bacteriological research laboratories and does not require column purification or other complex biochemical procedures. It should be useful to any laboratory which requires small amounts of lysin to produce L-forms and protoplasts or to release streptococcal antigens.


The cholinergic system in the pathophysiology and treatment of Alzheimer's disease.
Cholinergic synapses are ubiquitous in the human central nervous system. Their high density in the thalamus, striatum, limbic system, and neocortex suggest that cholinergic transmission is likely to be critically important for memory, learning, attention and other higher brain functions. Several lines of research suggest additional roles for cholinergic systems in overall brain homeostasis and plasticity. As such, the brain's cholinergic system occupies a central role in ongoing research related to normal cognition and age-related cognitive decline, including dementias such as Alzheimer's disease. The cholinergic hypothesis of Alzheimer's disease centres on the progressive loss of limbic and neocortical cholinergic innervation. Neurofibrillary degeneration in the basal forebrain is believed to be the primary cause for the dysfunction and death of forebrain cholinergic neurons, giving rise to a widespread presynaptic cholinergic denervation. Cholinesterase inhibitors increase the availability of acetylcholine at synapses in the brain and are one of the few drug therapies that have been proven clinically useful in the treatment of Alzheimer's disease dementia, thus validating the cholinergic system as an important therapeutic target in the disease. This review includes an overview of the role of the cholinergic system in cognition and an updated understanding of how cholinergic deficits in Alzheimer's disease interact with other aspects of disease pathophysiology, including plaques composed of amyloid-β proteins. This review also documents the benefits of cholinergic therapies at various stages of Alzheimer's disease and during long-term follow-up as visualized in novel imaging studies. The weight of the evidence supports the continued value of cholinergic drugs as a standard, cornerstone pharmacological approach in Alzheimer's disease, particularly as we look ahead to future combination therapies that address symptoms as well as disease progression.


Time series analysis of temporal trends in the pertussis incidence in Mainland China from 2005 to 2016.
Short-term forecast of pertussis incidence is helpful for advanced warning and planning resource needs for future epidemics. By utilizing the Auto-Regressive Integrated Moving Average (ARIMA) model and Exponential Smoothing (ETS) model as alterative models with R software, this paper analyzed data from Chinese Center for Disease Control and Prevention (China CDC) between January 2005 and June 2016. The ARIMA (0,1,0)(1,1,1)12 model (AICc = 1342.2 BIC = 1350.3) was selected as the best performing ARIMA model and the ETS (M,N,M) model (AICc = 1678.6, BIC = 1715.4) was selected as the best performing ETS model, and the ETS (M,N,M) model with the minimum RMSE was finally selected for in-sample-simulation and out-of-sample forecasting. Descriptive statistics showed that the reported number of pertussis cases by China CDC increased by 66.20% from 2005 (4058 cases) to 2015 (6744 cases). According to Hodrick-Prescott filter, there was an apparent cyclicity and seasonality in the pertussis reports. In out of sample forecasting, the model forecasted a relatively high incidence cases in 2016, which predicates an increasing risk of ongoing pertussis resurgence in the near future. In this regard, the ETS model would be a useful tool in simulating and forecasting the incidence of pertussis, and helping decision makers to take efficient decisions based on the advanced warning of disease incidence.


Look who's talking! Facial appearance can bias source monitoring.
When we see a stranger's face we quickly form impressions of his or her personality, and expectations of how the stranger might behave. Might these intuitive character judgements bias source monitoring? Participants read headlines "reported" by a trustworthy- and an untrustworthy-looking reporter. Subsequently, participants recalled which reporter provided each headline. Source memory for likely-sounding headlines was most accurate when a trustworthy-looking reporter had provided the headlines. Conversely, source memory for unlikely-sounding headlines was most accurate when an untrustworthy-looking reporter had provided the headlines. This bias appeared to be driven by the use of decision criteria during retrieval rather than differences in memory encoding. Nevertheless, the bias was apparently unrelated to variations in subjective confidence. These results show for the first time that intuitive, stereotyped judgements of others' appearance can bias memory attributions analogously to the biases that occur when people receive explicit information to distinguish sources. We suggest possible real-life consequences of these stereotype-driven source-monitoring biases.


Probiotic treatment by indigenous bacteria decreases mortality without disturbing the natural microbiota of Salvelinus fontinalis.
Next-generation sequencing is revealing the complex interactive networks of host-bacteria interactions, as it is now possible to screen in detail the microbiota harbored by a host. This study investigated the influence of a probiotic treatment on the survival and microbiota of brook charr (Salvelinus fontinalis), focusing on its disturbance of the natural microbiota (dysbiosis). The results indicated that an indigenous probiotic strain (identified as Rhodococcus sp.) colonized neither the fish skin mucus nor the water following the probiotic treatment. Instead, the probiotic strain was detected only in the biofilm of the test tank. Nevertheless, a substantial beneficial effect of the probiotic treatment was observed: the population of the pathogen Flavobacterium psychrophilum decreased in the treated tank water. This study clearly shows that the indigenous strain chosen for the probiotic treatment did not disturb the natural fish skin mucus microbiota but acted directly through the production system to control the growth of the pathogen and, as a consequence, to enhance fish survival.


Isolation and characterization of a plasmid from phase I Coxiella burnetii.
The DNA from the Nine Mile phase I strain of Coxiella burnetti, the etiological agent of Q fever, has been isolated and purified by cesium chloride-ethidium bromide density gradient centrifugation. A fraction of this DNA has a density characteristic of plasmid DNA. The plasmid DNA was cut with 20 different restriction endonucleases and shown to be a discrete entity. The plasmid, designated QpH1, is approximately 36 kilobases in size and has a molecular mass of 2.4 x 10(7) daltons. A partial restriction map of QpH1 has been constructed by using the restriction endonucleases SalI, KpnI, PstI, and XbaI. QpH1 DNA radioactively labeled by nick translation was used to show that sequences similar to the plasmid are also present in the phase II antigenic variant of C. burnetii.