Mammalian Golgi-associated Bicaudal-D2 functions in the dynein-dynactin pathway by interacting with these complexes.
Genetic analysis in Drosophila suggests that Bicaudal-D functions in an essential microtubule-based transport pathway, together with cytoplasmic dynein and dynactin. However, the molecular mechanism underlying interactions of these proteins has remained elusive. We show here that a mammalian homologue of Bicaudal-D, BICD2, binds to the dynamitin subunit of dynactin. This interaction is confirmed by mass spectrometry, immunoprecipitation studies and in vitro binding assays. In interphase cells, BICD2 mainly localizes to the Golgi complex and has properties of a peripheral coat protein, yet it also co-localizes with dynactin at microtubule plus ends. Overexpression studies using green fluorescent protein-tagged forms of BICD2 verify its intracellular distribution and co-localization with dynactin, and indicate that the C-terminus of BICD2 is responsible for Golgi targeting. Overexpression of the N-terminal domain of BICD2 disrupts minus-end-directed organelle distribution and this portion of BICD2 co-precipitates with cytoplasmic dynein. Nocodazole treatment of cells results in an extensive BICD2-dynactin-dynein co-localization. Taken together, these data suggest that mammalian BICD2 plays a role in the dynein- dynactin interaction on the surface of membranous organelles, by associating with these complexes.


Sensibility and taste alterations after impacted lower third molar extractions. A prospective cohort study.
To determine the incidence, severity and duration of lingual tactile and gustatory function impairments after lower third molar removal. Prospective cohort study with intra-subject measures of 16 patients undergoing lower third molar extractions. Sensibility and gustatory functions were evaluated in each subject preoperatively, one week and one month after the extraction, using Semmes-Weinstein monofilaments and 5 different concentrations of NaCl, respectively. Additionally, all patients filled a questionnaire to assess subjective perceptions. Although patients did not perceive any sensibility impairments, a statistically significant decrease was detected when Semmes-Weinstein monofilaments. This alteration was present at one week after the surgical procedure and fully recovered one month after the extraction. There were no variations regarding the gustatory function. Lower third molar removal under local anesthesia may cause light lingual sensibility impairment. Most of these alterations remain undetected to patients. These lingual nerve injuries are present one week after the extraction and recover one month after surgery. The taste seems to remain unaffected after these procedures.


Generalizability theory for the perplexed: a practical introduction and guide: AMEE Guide No. 68.
Generalizability theory (G theory) is a statistical method to analyze the results of psychometric tests, such as tests of performance like the Objective Structured Clinical Examination, written or computer-based knowledge tests, rating scales, or self-assessment and personality tests. It is a generalization of classical reliability theory, which examines the relative contribution of the primary variable of interest, the performance of subjects, compared to error variance. In G theory, various sources of error contributing to the inaccuracy of measurement are explored. G theory is a valuable tool in judging the methodological quality of an assessment method and improving its precision. Starting from basic statistical principles, we gradually develop and explain the method. We introduce tools to perform generalizability analysis, and illustrate the use of generalizability analysis with a series of common, practical examples in educational practice. We realize that statistics and mathematics can be either boring or fearsome to many physicians and educators, yet we believe that some foundations are necessary for a better understanding of generalizability analysis. Consequently, we have tried, wherever possible, to keep the use of equations to a minimum and to use a conversational and slightly "off-serious" style.


Radiomic biomarkers informative of cancerous transformation in neurofibromatosis-1 plexiform tumors.
This study explores whether objective, quantitative radiomic biomarkers derived from magnetic resonance (MR), positron emission tomography (PET), and computed tomography (CT) may be useful in reliably distinguishing malignant peripheral nerve sheath tumors (MPNST) from benign plexiform neurofibromas (PN). A registration and segmentation pipeline was established using a cohort of NF1 patients with histopathological diagnosis of PN or MPNST, and medical imaging of the PN including MR and PET-CT. The corrected MR datasets were registered to the corresponding PET-CT via landmark-based registration. PET standard-uptake value (SUV) thresholds were used to guide segmentation of volumes of interest: MPNST-associated PET-hot regions (SUV≥3.5) and PN-associated PET-elevated regions (2.0<SUV<3.5). Quantitative imaging features were extracted from the MR, PET, and CT data and compared for statistical differences. Intensity histogram features included (mean, media, maximum, variance, full width at half maximum, entropy, kurtosis, and skewness), while image texture was quantified using Law's texture energy measures, grey-level co-occurrence matrices, and neighborhood grey-tone difference matrices. For each of the 20 NF1 subjects, a total of 320 features were extracted from the image data. Feature reduction and statistical testing identified 9 independent radiomic biomarkers from the MR data (4 intensity and 5 texture) and 4 PET (2 intensity and 2 texture) were different between the PET-hot versus PET-elevated volumes of interest. Our data suggests imaging features can be used to distinguish malignancy in NF1-realted tumors, which could improve MPNST risk assessment and positively impact clinical management of NF1 patients.


Alzheimer's disease legislation and policy--now and in the future.
Recent studies have pointed to the large and increasingly complex issues surrounding dementia in American society in general and health care in particular. The initial foray into the federal policy arena, the National Alzheimer's Project Act, is a good first step but remains limited in scope and resources. Seeing the need for greater effort, thirty-three states have convened advisory groups and published their own plans for coordinating state-level activity to address Alzheimer's disease across service systems. This article examines the current federal and state policy statements on dementia-related issues and offers an outlook and suggestions for next steps along with the imperative for action on a significant scale. This is necessary to address issues in a meaningful way today instead of simply holding out hope for a treatment or cure in the future. Dementia in general and Alzheimer's disease in particular affect individuals and every facet of families, health care, business, states, and communities. While public policy statements at all levels call for increased coordination of resources, better communication, awareness, and essential linkages, more effort is required.


Bone morphogenetic protein-6 production in human osteoblastic cell lines. Selective regulation by estrogen.
Bone morphogenetic proteins (BMPs) induce differentiation of osteoblast and chondroblast lineage cells from uncommitted mesenchymal precursors. Because estrogen has potent osteochondrogenic actions, we investigated its effect on BMP production in two estrogen-responsive, human immortalized cell lines (hFOB/ER3 and hFOB/ER9) that display the mature osteoblast phenotype. These cell lines were produced by stable transfection of the estrogen receptor (ER) gene into immortalized fetal osteoblasts at low ( approximately 800 ER/ nucleus) and at high ( approximately 3, 900 ER/nucleus) levels, respectively. As assessed by reverse transcriptase PCR, treatment with 17beta-estradiol (10(-)10 - 10(-)7 M) increased steady-state levels of BMP-6 mRNA dose dependently by twofold in the hFOB/ER3 cells and by over threefold in the hFOB/ER9 cells. Messenger RNA levels for transforming growth factors-beta1 and -beta2 and BMPs-1 through -5 and -7 levels were unchanged. The results were confirmed by sequence determination of the PCR product and by Northern blot analysis for total RNA. 17beta-estradiol increased BMP-6 protein production sixfold by Western analysis. Cotreatment with antiestrogens (ICI 182,780 or 4-hydroxytamoxifen) antagonized the effects of 17beta-estradiol. These data suggest that some of the skeletal effects of estrogen on bone and cartilage may be mediated by increased production of BMP-6 by osteoblasts.


Studying N-linked glycosylation of receptor tyrosine kinases.
Metabolic alterations have been identified as a frequent event in cancer. This is often associated with increased flux through glycolysis, and also a secondary pathway to glycolysis, hexosamine biosynthetic pathway (HBP). HBP provides substrate for N-linked glycosylation, which occurs in the endoplasmic reticulum and the Golgi apparatus. N-linked glycosylation supports protein folding and correct sorting of proteins to plasma membrane and secretion. This process generates complex glycoforms, which can be recognized by other proteins and glycosylation of receptor tyrosine kinases (RTK) can also regulate their plasma-membrane retention time. Of special interest for experimental biologists, plants produce proteins, termed lectins, which bind with high specificity to glyco-conjugates. For the purposes of molecular biology, plant lectins can be conjugated to different moieties, such as agarose beads, which enable precipitation of specifically glycosylated proteins. In this chapter, we describe in detail how to perform pull-down experiments with commercially available lectins to identify changes in the glycosylation of RTKs.


Predicting extubation failure in blunt trauma patients with pulmonary contusion.
The need for reintubation after weaning from mechanical ventilation (extubation failure) is associated with increased morbidity and mortality. In blunt trauma patients with pulmonary contusion, factors predicting successful weaning have not been reliably defined. The purpose of this study was to identify criteria predicting successful extubation in these patients. Retrospective review during a 10-year period at a Level 1 trauma center was performed. A total of 173 extubations in 163 blunt trauma patients with pulmonary contusion requiring mechanical ventilation. Exclusion criteria include Glasgow Coma Scale (GCS) score of less than 9T before extubation, successful use of noninvasive positive-pressure ventilation after extubation, quadriplegia, and preextubation FIO2 of greater than 0.5. Data included age, Injury Severity Score (ISS), ventilator days, as well as GCS score, FIO2, the ratio of arterial oxygen tension to FIO2 (P/F ratio), and alveolar-arterial oxygen (A-a) difference at the time of extubation. Failure was defined as reintubation within 72 hours (excluding stridor or acute decline in GCS score). Mann-Whitney U-test, χ2 analysis, and logistic regression analysis determined variables associated with extubation failure. Odds ratios were used to compare P/F and A-a values associated with failed extubation. A total of 147 extubations (85%) were successful; 26 required reintubation. Patients did not differ by ISS, chest Abbreviated Injury Scale (AIS) score, presence of sternal or rib fractures, and admission pneumothorax or hemothorax. Increased age, A-a difference (≥ 120 mm Hg), and decreased P/F (<280) were associated with reintubation (p < 0.0001). By logistic regression analysis, P/F and A-a were independent variables for failed extubation; both remained independent risk factors when adjusted for age, ventilator days, GCS score, and preextubation FIO2. Using receiver operating characteristic curve inflection points for both P/F and A-a difference (area under the curve of 0.8 for both), patients with a P/F ratio less than 290 and an A-a difference of 100 mm Hg or greater were more likely to fail extubation (odds ratio, 9.2 and 8.7, respectively, p < 0.001). Blunt trauma patients with pulmonary contusion who are likely to fail extubation can be reliably identified using the readily available criteria of P/F ratio less than 290 and A-a difference of 100 mm Hg or greater.


Abnormal IGF-Binding Protein Profile in the Bone Marrow of Multiple Myeloma Patients.
Insulin-like growth factor (IGF) signalling plays a key role in homing, progression, and treatment resistance in multiple myeloma (MM). In the extracellular environment, the majority of IGF molecules are bound to one of six IGF-binding proteins (IGFBP1-6), leaving a minor fraction of total IGF free and accessible for receptor activation. In MM, high IGF-receptor type 1 expression levels correlate with a poor prognosis, but the status and role of IGF and IGFBPs in the pathobiology of MM is unknown. Here we measured total IGF1, IGF2, and intact IGFBP levels in blood and bone marrow samples from MM (n = 17), monoclonal gammopathy of undetermined significance (MGUS) (n = 37), and control individuals (n = 15), using ELISA (IGFs) and 125I-IGF1 Western Ligand Blotting (IGFBPs). MGUS and MM patients displayed a significant increase in intact IGFBP-2 (2.5-3.8 fold) and decrease in intact IGFBP-3 (0.6-0.5 fold) in the circulation compared to control individuals. Further, IGFBP-2 as well as total IGFBP levels were significantly lower in bone marrow compared to circulation in MM and MGUS only, whereas IGF1, IGF2, and IGFBP-3 were equally distributed between the two compartments. In conclusion, the profound change in IGFBP profile strongly suggests an increased IGF bioavailability in the bone marrow microenvironment in MGUS and MM, despite no change in growth factor concentration.


End-to-end sensor simulation for spectral band selection and optimization with application to the Sentinel-2 mission.
An end-to-end sensor simulation is a proper tool for the prediction of the sensor's performance over a range of conditions that cannot be easily measured. In this study, such a tool has been developed that enables the assessment of the optimum spectral resolution configuration of a sensor based on key applications. It employs the spectral molecular absorption and scattering properties of materials that are used for the identification and determination of the abundances of surface and atmospheric constituents and their interdependence on spatial resolution and signal-to-noise ratio as a basis for the detailed design and consolidation of spectral bands for the future Sentinel-2 sensor. The developed tools allow the computation of synthetic Sentinel-2 spectra that form the frame for the subsequent twofold analysis of bands in the atmospheric absorption and window regions. One part of the study comprises the assessment of optimal spatial and spectral resolution configurations for those bands used for atmospheric correction, optimized with regard to the retrieval of aerosols, water vapor, and the detection of cirrus clouds. The second part of the study presents the optimization of thematic bands, mainly driven by the spectral characteristics of vegetation constituents and minerals. The investigation is performed for different wavelength ranges because most remote sensing applications require the use of specific band combinations rather than single bands. The results from the important "red-edge" and the "short-wave infrared" domains are presented. The recommended optimum spectral design predominantly confirms the sensor parameters given by the European Space Agency. The system is capable of retrieving atmospheric and geobiophysical parameters with enhanced quality compared to existing multispectral sensors. Minor spectral changes of single bands are discussed in the context of typical remote sensing applications, supplemented by the recommendation of a few new bands for the next generation of optical Sentinel sensors.


Scaling Tableting Processes from Compaction Simulator to Rotary Presses-Mind the Sub-Processes.
Compaction simulators are frequently used in the formulation and process development of tablets, bringing about the advantages of flexibility, low material consumption, and high instrumentation to generate the most possible process understanding. However, their capability of resembling general aspects of rotary press compaction and their precision in simulating or mimicking sub-processes such as feeding and filling need to be systematically studied. The effect of material deformation behavior, blend composition, and feeding on tensile strength and simulation precision as compared with rotary presses of different scales is evaluated in this study. Generally, good simulation performance was found for the studied compaction simulator. Compaction profile-sensitivity was demonstrated for highly visco-plastic materials while shear-sensitivity in feeding was demonstrated for lubricated blends of ductile particles. Strategies for the compensation of both in compaction simulator experiments are presented by careful investigation of the compaction stress over time profiles and introduction of a compaction simulator-adapted shear number approach to account for differences in layout and operation mode between compaction simulator and rotary press, respectively. These approaches support the general aim of this study to provide a more straightforward determination of scaling process parameters between rotary press and compaction simulator and facilitate a quicker and more reliable process transfer.


Increased immunosuppression, not anticoagulation, extends cardiac xenograft survival.
Cardiac xenograft function is lost due to delayed xenograft rejection (DXR) characterized by microvascular thrombosis and myocardial necrosis. The cause of DXR is unknown but may result from thrombosis induced by antibody-mediated activation of endothelial cells and/or by incompatibilities in thromboregulatory interactions. To examine these issues, a series (Groups 1-6) of previous transgenic CD46 pig-to-baboon heterotopic cardiac transplants were reanalyzed for baseline immunosuppressive levels, graft survival and infectious complications with and without systemic anticoagulation. Groups 1-4 received low dose tacrolimus and sirolimus maintenance therapy, with splenectomy, anti-CD20 and daily alpha-Gal polymer. Group 1 recipients received no anticoagulation. Groups 2-4 were anticoagulated with aspirin and Plavix, Lovenox, or Coumadin, respectively. Group 5 was treated with Lovenox and high dose tacrolimus and sirolimus maintenance therapy. Group 6 recipients received no postoperative anticoagulation but the same immunosuppression as group 5. Median survival (15-22 days) within groups 1-4 was not significantly different. At rejection all tissues exhibited microvascular thrombosis, coagulative necrosis and similar levels of platelet and fibrin deposition. Groups 5 and 6 median survival (76 days) was significantly increased compared to groups 1-4. There was no significant difference in median survival between Lovenox treated recipients (68 days) and anticoagulant free recipients (96 days). Rejected tissues showed vascular antibody deposition, microvascular thrombosis, and myocyte necrosis. Significant prolongation in xenograft survival is achieved by improved immunosuppression. These results suggest that ongoing immune responses remain the major stimulus for DXR.


Beta-amyloid precursor protein mutants respond to gamma-secretase modulators.
Pathogenic generation of the 42-amino acid variant of the amyloid beta-peptide (Abeta) by beta- and gamma-secretase cleavage of the beta-amyloid precursor protein (APP) is believed to be causative for Alzheimer disease (AD). Lowering of Abeta(42) production by gamma-secretase modulators (GSMs) is a hopeful approach toward AD treatment. The mechanism of GSM action is not fully understood. Moreover, whether GSMs target the Abeta domain is controversial. To further our understanding of the mode of action of GSMs and the cleavage mechanism of gamma-secretase, we analyzed mutations located at different positions of the APP transmembrane domain around or within the Abeta domain regarding their response to GSMs. We found that Abeta(42)-increasing familial AD mutations of the gamma-secretase cleavage site domain responded robustly to Abeta(42)-lowering GSMs, especially to the potent compound GSM-1, irrespective of the amount of Abeta(42) produced. We thus expect that familial AD patients carrying mutations at the gamma-secretase cleavage sites of APP should respond to GSM-based therapeutic approaches. Systematic phenylalanine-scanning mutagenesis of this region revealed a high permissiveness to GSM-1 and demonstrated a complex mechanism of GSM action as other Abeta species (Abeta(41), Abeta(39)) could also be lowered besides Abeta(42). Moreover, certain mutations simultaneously increased Abeta(42) and the shorter peptide Abeta(38), arguing that the proposed precursor-product relationship of these Abeta species is not general. Finally, mutations of residues in the proposed GSM-binding site implicated in Abeta(42) generation (Gly-29, Gly-33) and potentially in GSM-binding (Lys-28) were also responsive to GSMs, a finding that may question APP substrate targeting of GSMs.


First Report of Strawberry necrotic shock virus in China.
Strawberry necrotic shock virus (SNSV) is an economically important viral pathogen that infects Fragaria and Rubus spp. SNSV was first identified in the 1950s and early studies indicated that SNSV was a strain of Tobacco streak virus (TSV). Recently, it was shown that SNSV was a distinct virus based on molecular characterization (2). Currently, SNSV is a tentative member of the Ilarvirus genus in the Bromoviridae family. In 2008, a small sampling survey for SNSV was done in Heilongjiang Province of China, and 15 strawberry samples were collected from symptomless strawberries in a home garden that had more than 5 years of strawberry cultivation history. Total nucleic acid was extracted from strawberry leaflets by modified cetyltrimethylammoniumbromide methods (3). Reverse transcription (RT)-PCR was operated with the published primer pair CPbeg F/CPend R (2). Amplified DNA fragments with the predicted size were obtained only in one strawberry sample, which was further cloned and sequenced. The sequence (GenBank Accession No. HQ830017) was closely related and highly homologous (89.7 to 98.5% identity) to that of viral isolates (GenBank Accession Nos. AY363228-AY363242) from Fragaria and Rubus spp. Phylogenetic analysis based on nucleotide sequence of the coat protein gene was done with the neighbor-joining method of MEGA 4.0 software. The result showed that all the isolates of SNSV fell into two distinct clades. The Chinese isolate formed one small clade with Japanese isolate 1291. The isolate was also transmitted to Chenopodium quinoa by mechanical inoculation in the greenhouse, and the symptom of chlorotic mottling could be found in C. quinoa and detected by RT-PCR. To determine whether the sample was infected by other strawberry viruses, RT-PCR assays with the published primer pairs SVBVdeta/SVBVdetb, SMoVdeta/SMoVdetb, and SMYEVdeta/SMYEVdetb were also performed for detection of Strawberry vein banding virus, Strawberry mottle virus, and Strawberry mild yellow edge virus using total nucleic acid extracted from the SNSV-positive sample as a template (1). The result indicated that it had been also infected by Strawberry mild yellow edge virus, although no visible symptoms were observed. To our knowledge, this is the first report of SNSV in strawberry in China. Additional work is needed to elucidate the biological characterization and significance of the finding. References: (1) J. R. Thompson et al. J. Virol. Methods 111:85, 2003. (2) I. E. Tzanetakis et al. Arch. Virol. 149:2001, 2004. (3) H. Y. Yang et al. Acta Hortic. 764:127, 2007.


SIRT1 and SIRT3 deacetylate homologous substrates: AceCS1,2 and HMGCS1,2.
SIRT1 and SIRT3 are NAD+-dependent protein deacetylases that are evolutionarily conserved across mammals. These proteins are located in the cytoplasm/nucleus and mitochondria, respectively. Previous reports demonstrated that human SIRT1 deacetylates Acetyl-CoA Synthase 1 (AceCS1) in the cytoplasm, whereas SIRT3 deacetylates the homologous Acetyl-CoA Synthase 2 (AceCS2) in the mitochondria. We recently showed that 3-hydroxy-3-methylglutaryl CoA synthase 2 (HMGCS2) is deacetylated by SIRT3 in mitochondria, and we demonstrate here that SIRT1 deacetylates the homologous 3-hydroxy-3-methylglutaryl CoA synthase 1 (HMGCS1) in the cytoplasm. This novel pattern of substrate homology between cytoplasmic SIRT1 and mitochondrial SIRT3 suggests that considering evolutionary relationships between the sirtuins and their substrates may help to identify and understand the functions and interactions of this gene family. In this perspective, we take a first step by characterizing the evolutionary history of the sirtuins and these substrate families.


A calcineurin-dependent transcriptional pathway for cardiac hypertrophy.
In response to numerous pathologic stimuli, the myocardium undergoes a hypertrophic response characterized by increased myocardial cell size and activation of fetal cardiac genes. We show that cardiac hypertrophy is induced by the calcium-dependent phosphatase calcineurin, which dephosphorylates the transcription factor NF-AT3, enabling it to translocate to the nucleus. NF-AT3 interacts with the cardiac zinc finger transcription factor GATA4, resulting in synergistic activation of cardiac transcription. Transgenic mice that express activated forms of calcineurin or NF-AT3 in the heart develop cardiac hypertrophy and heart failure that mimic human heart disease. Pharmacologic inhibition of calcineurin activity blocks hypertrophy in vivo and in vitro. These results define a novel hypertrophic signaling pathway and suggest pharmacologic approaches to prevent cardiac hypertrophy and heart failure.


Antibiotic resistance--what's dosing got to do with it?
This review seeks to identify original research articles that link antibiotic dosing and the development of antibiotic resistance for different antibiotic classes. Using this data, we seek to apply pharmacodynamic principles to assist clinical practice for suppressing the emergence of resistance. Concepts such as mutant selection window and mutant prevention concentration will be discussed. PubMed, EMBASE, and the Cochrane Controlled Trial Register. All articles that related antibiotic doses and exposure to the formation of antibiotic resistance were reviewed. The escalation of antibiotic resistance continues worldwide, most prominently in patients in intensive care units. Data are emerging from in vitro and in vivo studies that suggest that inappropriately low antibiotic dosing may be contributing to the increasing rate of antibiotic resistance. Fluoroquinolones have widely been researched and publications on other antibiotic classes are emerging. Developing dosing regimens that adhere to pharmacodynamic principles and maximize antibiotic exposure is essential to reduce the increasing rate of antibiotic resistance. Antibiotic dosing must aim to address not only the bacteria isolated, but also the most resistant subpopulation in the colony, to prevent the advent of further resistant infections because of the inadvertent selection pressure of current dosing regimens. This may be achieved by maximizing antibiotic exposure by administering the highest recommended dose to the patient.


Separate mechanisms activate sigma B of Bacillus subtilis in response to environmental and metabolic stresses.
sigma B is a secondary sigma factor that controls the general stress response of Bacillus subtilis. sigma B-dependent transcription is induced by the activation of sigma B itself, a process that involves release of sigma B from an inhibitory complex with its primary regulator, RsbW. sigma B becomes available to RNA polymerase when RsbW forms a complex with an additional regulatory protein (RsbV) and, because of this, fails to bind sigma B. Using Western blot (immunoblot) analyses, reporter gene fusion assays, and measurements of nucleotide pool sizes, we provide evidence for two independent processes that promote the binding of RsbW to RsbV. The first occurs during carbon limitation or entry into stationary phase. Activation of sigma B under these circumstances correlates with a drop in the intracellular levels of ATP and may be a direct consequence of ATP levels on RsbW's binding preference. The second activation process relies on the product of a third regulatory gene, rsbU. RsbU is dispensable for sigma B activation during carbon limitation or stationary phase but is needed for activation of sigma B in response to any of a number of different environmental insults (ethanol treatment, salt or acid shock, etc.). RsbU, or a process dependent on it, alters RsbW binding without regard for intracellular levels of ATP. In at least some instances, the effects of multiple inducing stimuli are additive. The data are consistent with RsbW being a regulator at which distinct signals from separate effectors can be integrated to modulate sigma B activity.


Life lost due to premature deaths in New South Wales, Australia.
This study attempts to measure premature mortality, in addition to overall death rates, in order to provide more information that can be used to develop and monitor health programmes that are aimed at reducing premature (often preventable) mortality in New South Wales (NSW), Australia. Premature years of potential life lost (PYPLL) and valued years of potential life lost methods are applied for mortality data in NSW from 1990 to 2002. Variations in these measures for 2001 are studied further in terms of age, sex, urban/rural residence, and socio-economic status. PYPLL rates for all leading causes of death have declined. It is shown that the average male to female ratio of PYPLLs is highest for accidents, injury and poisoning (3.4:1) followed by mental disorders (2.7:1) and cardiovascular diseases (2.6:1). Although fewer women than men die of cardiovascular diseases, there is a greater proportionate importance of cerebrovascular mortality among women. In order to further reduce premature deaths, programs are required to improve the health of people living in lower socio-economic status areas, especially in rural NSW. Targeted regional or community level programs are required to reduce avoidable deaths due to accidents, injury and poisoning occasioned by motor vehicle accidents, poisoning and suicide among young adults.


Tick histamine-binding proteins: lipocalins with a second binding cavity.
Tick histamine-binding proteins (HBPs) are lipocalins with two binding pockets. One of these binds histamine with a high affinity and is found at the position expected from other lipocalins, adjacent to the omega-loop at the open-end of the beta-barrel. A second binding cavity, which is a low-affinity site for histamine in one of the HBPs, is located at the end of the barrel that is closed off in other lipocalins. In order to create the second site, the 'closed-end' region has undergone a major reconstruction. Typical lipocalin characteristics, such as the 3(10) helix and a structural cluster of highly conserved residues, have been lost, while an alpha-helix now shields the cavity from the exterior. The prominence of acidic residues in the binding pockets is another distinctive characteristic of HBPs. Whereas most lipocalins have highly hydrophobic binding cavities designed to bind lipophilic compounds, HBPs have evolved to trap cationic, hydrophilic molecules.


Intra-articular transplantation of atsttrin-transduced mesenchymal stem cells ameliorate osteoarthritis development.
Osteoarthritis (OA) remains an intractable clinical challenge. Few drugs are available for reversing this degenerative disease, although some promising candidates have performed well in preclinical studies. Tumor necrosis factor α (TNFα) has been identified as a crucial effector modulating OA pathogenesis. This study aimed to investigate the therapeutic effects of Atsttrin, a novel TNFα blocker, on OA treatment. We developed genetically modified mesenchymal stem cells (MSCs) that expressed recombinant Atsttrin (named as MSC-Atsttrin). Expression levels of ADAMTS-5, MMP13, and iNOS of human chondrocytes were analyzed when cocultured with MSC-GFP/Atsttrin. OA animal models were induced by anterior cruciate ligament transection, and MSC-GFP/Atsttrin were injected into the articular cavity 1 week postsurgery. The results showed that MSC-Atsttrin significantly suppressed TNFα-driven up-regulation of matrix proteases and inflammatory factors. Intra-articular injection of MSC-Atsttrin prevented the progression of degenerative changes in the surgically induced OA mouse model. Additionally, levels of detrimental matrix hydrolases were significantly diminished. Compared with nontreated OA samples at 8 weeks postsurgery, the percentages of MMP13- and ADAMTS-5-positive cells were significantly reduced from 91.33% ± 9.87% to 24.33% ± 5.7% (p < .001) and from 91.33% ± 7.1% to 16.67% ± 3.1% (p < .001), respectively. Our results thus indicated that suppression of TNFα activity is an effective strategy for OA treatment and that intra-articular injection of MSCs-Atsttrin could be a promising therapeutic modality.


Instrumented Trail-Making Task: Application of Wearable Sensor to Determine Physical Frailty Phenotypes.
The physical frailty assessment tools that are currently available are often time consuming to use with limited feasibility. To address these limitations, an instrumented trail-making task (iTMT) platform was developed using wearable technology to automate quantification of frailty phenotypes without the need of a frailty walking test. Sixty-one older adults (age = 72.8 ± 9.9 years, body mass index [BMI] = 27.4 ± 4.9 kg/m2) were recruited. According to the Fried Frailty Criteria, 39% of participants were determined as robust and 61% as non-robust (pre-frail or frail). In addition, 17 young subjects (age = 29.0 ± 7.2 years, BMI = 26.2 ± 4.6 kg/m2) were recruited to determine the healthy benchmark. The iTMT included reaching 5 indexed circles (including numbers 1-to-3 and letters A&B placed in random orders), which virtually appeared on a computer-screen, by rotating one's ankle-joint while standing. By using an ankle-worn inertial sensor, 3D ankle-rotation was estimated and mapped into navigation of a computer-cursor in real-time (100 Hz), allowing subjects to navigate the computer-cursor to perform the iTMT. The ankle-sensor was also used for quantifying ankle-rotation velocity (representing slowness), its decline during the test (representing exhaustion), and ankle-velocity variability (representing movement inefficiency), as well as the power (representing weakness) generated during the test. Comparative assessments included Fried frailty phenotypes and gait assessment. All subjects were able to complete the iTMT, with an average completion time of 125 ± 85 s. The iTMT-derived parameters were able to identify the presence and absence of slowness, exhaustion, weakness, and inactivity phenotypes (Cohen's d effect size = 0.90-1.40). The iTMT Velocity was significantly different between groups (d = 0.62-1.47). Significant correlation was observed between the iTMT Velocity and gait speed (r = 0.684 p < 0.001). The iTMT-derived parameters and age together enabled significant distinguishing of non-robust cases with area under curve of 0.834, sensitivity of 83%, and specificity of 67%. This study demonstrated a non-gait-based wearable platform to objectively quantify frailty phenotypes and determine physical frailty, using a quick and practical test. This platform may address the hurdles of conventional physical frailty phenotypes methods by replacing the conventional frailty walking test with an automated and objective process that reduces the time of assessment and is more practical for those with mobility limitations.


The clinical epidemiology of childhood pulmonary tuberculosis: a critical review of literature from the pre-chemotherapy era.
The pre-chemotherapy literature represents an impressive body of evidence that clarifies important epidemiological concepts in childhood tuberculosis. Reports describe the major transitions in tuberculosis, from exposure to infection and from infection to disease (morbidity and mortality), without the influence of chemotherapy. Children with household exposure to a sputum smear-positive source case experienced the greatest risk of becoming infected and of developing subsequent disease. Household exposure to a sputum smear-negative source case or non-household exposure still posed an appreciable, although greatly reduced, risk. Infection in children less than 2 years of age indicated a probable household source case. The majority of older children who were infected did not have a household source identified, and presumably became infected in the community. The annual risk of infection (ARI) was not constant across all ages, but seemed to increase during periods of widening social contact. Infants and adolescents were the groups at highest risk for disease development and death following primary infection.


A deceptive mass on neonatal ultrasound and a fetal brain MRI-confirmed complex dural arteriovenous fistula postnatally: illustrative case.
Dural arteriovenous fistulas (dAVFs) are direct, aberrant connections between dural arteries and cerebral veins. In neonates, delayed diagnosis results in grim outcomes. Treatment involves endovascular management because of its success and tolerability. Here, the authors present a case of a complex dAVF initially recognized with an in utero neurosonogram and fetal magnetic resonance imaging (MRI). A 21-week fetal ultrasound of a nonspecific brain mass was confirmed with fetal MRI as a 2.7-cm T1-hyperintense posterior fossa mass. Although a large flow void in the left middle cranial fossa was present, postnatal computed tomography angiography ultimately revealed a high-flow dAVF communicating with the left transverse sinus. In the early postnatal period, the patient developed hydrocephalus. After successful partial embolization, 6-week postangiogram brain MRI indicated disease progression with the development of a venous varix causing brainstem compression. Repeat embolization resulted in complete cessation of early venous drainage. Neonatal dAVFs are exceedingly rare and result in futile outcomes; however, detection in utero is possible. Although definitive therapy must be performed postnatally, constant monitoring and early delivery can prevent complications. Attention to fetal ultrasound is essential, and knowledge of fetal MRI in the detection of these complex lesions can significantly improve outcomes.


Address entry while driving: speech recognition versus a touch-screen keyboard.
A driving simulator experiment was conducted to determine the effects of entering addresses into a navigation system during driving. Participants drove on roads of varying visual demand while entering addresses. Three address entry methods were explored: word-based speech recognition, character-based speech recognition, and typing on a touch-screen keyboard. For each method, vehicle control and task measures, glance timing, and subjective ratings were examined. During driving, word-based speech recognition yielded the shortest total task time (15.3 s), followed by character-based speech recognition (41.0 s) and touch-screen keyboard (86.0 s). The standard deviation of lateral position when performing keyboard entry (0.21 m) was 60% higher than that for all other address entry methods (0.13 m). Degradation of vehicle control associated with address entry using a touch screen suggests that the use of speech recognition is favorable. Speech recognition systems with visual feedback, however, even with excellent accuracy, are not without performance consequences. Applications of this research include the design of in-vehicle navigation systems as well as other systems requiring significant driver input, such as E-mail, the Internet, and text messaging.


Morphometric data to FIGO stage and histological type and grade for prognosis of ovarian tumours.
The prognostic value of using histological typing, grading, and morphology, in addition to clinical staging, was assessed in 98 cases of invasive ovarian cancer of the common epithelial types (serous, mucinous, and endometrial). All of these cases had at least five years of follow up. When regression analysis was used, the International Federation of Gynaecology and Obstetrics' (FIGO) staging system was the best indicator for prognosis. Analysis of a combination of morphometric features was the second best indicator, being especially useful for the those patients with stage I disease. Variables that indicated a relatively poor prognostic outcome were mitotic index above 30; volume percentage epithelium above 65%; shortest nuclear axis above a mean of 1 X 1 micrometers. Histological typing of ovarian tumours was of limited value; mucinous tumours have a somewhat better prognosis than serous tumours, but the prognostic value of typing alone was found to be limited. Qualitative histological grading was useful, but the prognostic value of morphometric grading was better. Measurement of morphological features with an interactive computer program is simple and can be done by a pathologist or a technician: in future it is likely that such automated systems of measurement will improve the objectivity of tissue analysis.


Range of Motion Improvement Following Surgical Management of Knee Arthrofibrosis in Children and Adolescents.
Arthrofibrosis of the knee is well-described in adults as a potentially debilitating postoperative complication following anterior cruciate ligament reconstruction, total knee arthroplasty, or fracture fixation. Knee arthrofibrosis in children and adolescents, however, has received little attention. The primary purpose of this study was to report improvements in range of motion (ROM) following lysis of adhesions and manipulation under anesthesia (LOA/MUA) in children and adolescents with knee arthrofibrosis, and, secondarily, to evaluate for any effect of preoperative dynamic splinting on ROM outcomes. Ninety patients aged 18 years and below (mean, 14.4±3.5) and 31% male who underwent LOA/MUA at an urban tertiary care hospital following prior knee surgery were evaluated. Demographic, clinical, ROM, and revision data were compiled. Primary outcome was absolute ROM. Secondarily, ROM was analyzed as a categorical variable with "Full ROM" defined to be -5 to 130 degrees or better, "functional" ROM was defined as unable to obtain -5 to 130 degrees but not requiring revision, and "failure" defined as resulting in revision arthrofibrosis surgery. t tests and χ analyses were used to compare ROM and count variables between dynamic splinting subgroups. Mean time from index surgery to LOA/MUA was 6.0±4.4 months, and follow-up was 42±56 months. Index procedures included anterior cruciate ligament reconstruction (N=33), tibial spine arthroscopic reduction and internal fixation (N=18), fracture fixation (N=17), soft tissue repair (N=17), and multiligament reconstruction (N=5). In total, 68 subjects (76%) had any flexion loss, 57 subjects (63%) had any extension loss, and 40 subjects (44%) had both flexion and extension loss.Fifty-six subjects (62%) had full ROM at final follow-up, 25 subjects (28%) had functional ROM, and 9 subjects (10%) required revision. No demographic, clinical, or surgical variable was predictive of treatment failure. Patients who underwent dynamic splinting preoperatively (N=46; 51%) had greater preoperative flexion (99±16 vs.77±34 degrees; P=0.001), but no difference in flexion at final follow-up (121±20 vs.128±11 degrees; P=0.08). Failure was not associated with time from index procedure to LOA/MUA, and the proportion who regained full ROM postoperatively was equivalent between those who had dynamic splinting and those who did not (65% vs. 59%; P=0.70). LOA/MUA for children with arthrofibrotic knees results in significant improvements in ROM with 90% revision-free success. Preoperative dynamic or static progressive splinting improves preoperative flexion but does not affect postoperative range of motion or failure rate. Level II.


Right heart dysfunction after left ventricular assist device implantation: a comparison of the pulsatile HeartMate I and axial-flow HeartMate II devices.
Right heart dysfunction confers significant morbidity and mortality after left ventricular assist device implantation and historically occurs in as many as a third of patients. It is unknown whether newer axial flow pumps have a different impact on postimplant right heart dysfunction. We compared the incidence of right heart dysfunction after implantation of the pulsatile HeartMate I (XVE) and the continuous flow HeartMate II left ventricular assist device. We retrospectively reviewed patients who underwent HeartMate I or HeartMate II implantation between June 2000 and March 2007. Right heart dysfunction was defined as inotropic/vasodilator support for 14 or more consecutive days or the need for a right ventricular assist device, or both. Seventy-seven patients underwent HeartMate implantation; 43 received a HeartMate I and 34 received a HeartMate II, for a mean left ventricular assist device support time of 202 and 160 days, respectively. Operative mortality was lower for HeartMate II patients (28% versus 15%; p = 0.26). The HeartMate II patients had lower preoperative right ventricular stroke work index. Pulmonary vascular resistance index, right ventricular stroke work index, and pulmonary and right atrial pressures improved and were similar between groups postoperatively. Overall, right heart dysfunction developed in 35% of HeartMate I patients (15 of 43) and 41% of HeartMate II patients (14 of 34; p = 0.63). Fewer HeartMate II patients (2) than HeartMate I patients (5) required 7 or more days of epinephrine, whereas more HeartMate II patients (7) than HeartMate I patients (5) required 7 or more days of milrinone. Six HeartMate I and 3 HeartMate II patients required right ventricular assist device implantation for right heart failure. Survival was similar (p = 0.7) between groups at, respectively, 3 (63% versus 62%), 6 (58% versus 58%), and 12 months (49% versus 48%). Right heart dysfunction is a persistent clinical problem after left ventricular assist device placement. We report the first study comparing the incidence of right heart dysfunction after HeartMate I versus HeartMate II implantation. Although the incidence of right heart dysfunction was similar, fewer HeartMate II patients required right ventricular assist device placement and fewer required pure inotropic support for right heart failure.


Colorful surface architectures with three different types of dynamic covalent bonds: integration of anthocyanins, tritylium ions and flavins.
Although they combine the best of covalent and non-covalent bonds, dynamic covalent bonds are usually not used together. Building on pioneering examples for functional systems with two orthogonal dynamic covalent bonds, we herein elaborate on multicomponent surface architectures that operate with three different types of dynamic covalent bonds. Disulfide exchange under basic conditions is used to grow single π stacks directly on oxide surfaces, hydrazone exchange under acidic conditions to add a second string or stack, and boronic-ester exchange under neutral conditions to build the third one. In this study, we show that this synthetic approach to complex systems provides access to emergent properties, as exemplified with ordered stacks of anthocyanins, pyrocatchol violet and riboflavins. The integration of anthocyanins, the central component of the pigments of plant flowers, is interesting to protect the blue flavylium cation against deprotonation, deplanarization and degradation. The integration of pyrocatchol violet is of interest to stabilize the blue, disfavored tritylium cation. The red riboflavin stacks are attractive because they generate high photocurrent. These colorful examples hint at the potential of synthetic methods that use three different types of dynamic covalent bonds in concert to build complex systems with emergent properties.


Cost-effectiveness of red blood cell transfusion vs. non-intervention in women with acute anaemia after postpartum haemorrhage.
Red blood cell (RBC) transfusion is frequently used to treat women with acute anaemia after postpartum haemorrhage. We aimed to assess the economic consequences of red blood cell transfusion compared to non-intervention in these women. A trial-based cost-effectiveness analysis was performed alongside the Well-Being of Obstetric patients on Minimal Blood transfusions (WOMB) trial. Women with acute anaemia [Hb 4·8-7·9 g/dl (3·0-4·9 mm)] after postpartum haemorrhage, without severe anaemic symptoms, were randomly allocated to RBC transfusion or non-intervention. Primary outcome of the trial was physical fatigue (Multidimensional Fatigue Inventory, scale 4-20; 20 represents maximal fatigue). Total costs per arm were calculated using a hospital perspective with a 6 weeks time horizon. Per woman, mean costs in the RBC transfusion arm (n = 258) were €1957 compared to €1708 in the non-intervention arm (n = 261; P = 0·024). The 13% difference in costs between study arms predominantly originated from costs of RBC units, as costs of RBC units were six times higher in the RBC transfusion arm. RBC transfusion led to a small improvement in physical fatigue of 0·58 points per day; thus, the costs to improve the physical fatigue score with one point would be €431. In women with acute anaemia after postpartum haemorrhage (PPH), RBC transfusion is on average €249 more expensive per woman than non-intervention, with only a small gain in HRQoL after RBC transfusion. Taking both clinical and economic consequences into account, implementation of a non-intervention policy seems justified.


Multi-scale multi-level marine spatial planning: A novel methodological approach applied in South Africa.
This study proposes and discusses a multi-scale spatial planning method implemented simultaneously at local and national level to prioritize ecosystem management actions across landscapes and seascapes. Mismatches in scale between the occurrence of biodiversity patterns and ecological processes, and the size and nature of the human footprint, and the different levels and scope of governance, are a significant challenge in conservation planning. These scale mismatches are further confounded by data resolution disparities across and amongst the different scales. To address this challenge, we developed a multi-resolution scale-linked marine spatial planning method. We tested this approach in the development of a Conservation Plan for a significant portion of South Africa's exclusive economic zone, adjacent to the east coast province of KwaZulu-Natal (the SeaPlan project). The study's dataset integrated the geographic distribution of 390 biodiversity elements (species, habitats, and oceanographic processes) and 38 human activities. A multi-resolution system of planning unit layers (PUL), with individual PUs ranging in resolution from 0.2 to 10 km, was designed to arrange and analyse these data. Spatial priorities for conservation were selected incrementally at different scales, contributing conservation targets from the fine-, medium- and large-scale analyses, and from the coast to the offshore. Compared to a basic single-resolution scale-unlinked plan, our multi-resolution scale-linked method selects 6% less conservation area to achieve the same targets. Compared to a multi-resolution scale-unlinked plan, our method requires only an additional 5% area. Overall, this method reflects the multi-scale nature of marine social-ecological systems more realistically, is relatively simple and replicable, and serves to better connect fine-scale and large-scale spatial management policies. We discuss the impacts of this study on protected area expansion planning processes in South Africa. This study showcases a methodological advance that has the potential to impact marine spatial planning practices and policies.


Comparison between automatic ribotyping and random amplified polymorphic DNA analysis of Bacillus cereus isolates from the dairy industry.
Discrimination by automatic ribotyping and random amplified polymorphic DNA PCR, RAPD, was compared for 40 different B. cereus dairy isolates, 4 different B. mycoides isolates and 6 culture collection strains. RAPD-PCR has previously shown to be useful for tracing contamination routes of B. cereus to milk. Automatic ribotyping using EcoRI and PvuII separated the B. cereus and B. mycoides isolates/strains into 36 different ribotypes. RAPD-typing with primers generated 40 different RAPD-profiles. However, 17 isolates clustered into eight groups, irrespective of the primer and restriction enzyme used, and in all but one case, the isolates with the same pattern were isolated from the same dairy. Automatic ribotyping proved to be a useful, standardized and quick method to discriminate between B. cereus strains, only slightly less discriminatory than RAPD-typing.


Antisocial process screening device: validation on a Russian sample of juvenile delinquents with the emphasis on the role of personality and parental rearing.
The objectives of the present study were 1) to validate the Antisocial Process Screening Device (APSD) in a sample of Russian juvenile delinquents; 2) to examine subgroups of delinquents with higher versus lower levels of childhood problem behaviors with respect to the APSD subscales, personality traits, and parental rearing; and 3) to attempt to replicate the previous finding that the APSD subscale measuring callous/unemotional traits can differentiate subgroups of delinquents with different precursors for problem behaviors (predominantly biological versus predominantly social). A group of 250 Russian juvenile inmates (mean age=16.4) was examined by means of the APSD completed by the staff at the correctional institution. The inmates completed several self-reports assessing their current and childhood behavior problems, personality traits and experienced parental rearing practices. A factor structure of the APSD was obtained that is similar, albeit not identical, to that from the original studies by Frick and colleagues [Frick, P.J., O'Brien, B.S., Wootton, J.M., McBurnett, K., (1994). Psychopathy and conduct problems in children. Journal of Abnormal Psychology, 103, 700-707]; [Frick, P.J., Barry, C.T., Bodin, S.D., (1999). Applying the concept of psychopathy to children: Implications for the Assessment of antisocial youth. In Gacono, C.B. (Ed), The clinical and forensic assessment of psychopathy: A practitioners guide. Hillsdale, NJ: Erlbaum]; [Frick, P.J., Lilienfeld, S.O., Ellis, M., Loney, B., Silverthorn, P., (1999). The association between anxiety and psychopathy dimensions in children. Journal of Abnormal Child Psychology, 27, 383-392]; callous unemotional traits in the present sample were expressed in manipulative behavior. Results further disclosed higher levels of antisocial and aggressive activities, higher levels of personality attributes such as narcissism and novelty seeking, as well as lower cooperativeness, and negatively perceived parental rearing in a subgroup with higher (versus lower) number of childhood symptoms of conduct disorder and oppositional disorder. The juvenile delinquents with higher levels as compared to lower levels of callous unemotional traits also perceived their parents as using more negative rearing strategies. The findings are discussed in terms of interactional processes between personality of the juvenile delinquents and parental rearing in the development of antisocial behavior.


Single- and multiple-dose pharmacokinetics, safety and tolerability of zibotentan (ZD4054) in Chinese men with advanced solid tumors.
The endothelin axis and the endothelin A (ET(A)) receptor have been implicated in tumor development and bone metastasis. This study aimed to investigate the pharmacokinetic (PK) and safety profiles of the specific ET(A) receptor antagonist, zibotentan, in elderly, male Chinese patients with advanced solid tumors. The PK data generated in these Chinese patients were further compared with those previously reported in Japanese and Caucasian patient populations. In this Phase I, open-label study, patients received a single dose of zibotentan 10 mg on Day 1, followed by a 72-h washout period and 12 consecutive days of once-daily zibotentan 10 mg. Fifteen patients received at least one dose of zibotentan 10 mg. Exposure was demonstrated in all patients and the PK profiles following single dosing and multiple dosing showed relatively rapid absorption, decline in a monophasic manner, a modest amount of accumulation, and relatively low apparent clearance and volume of distribution. Zibotentan was well tolerated with no new safety concerns. Adverse events reported in >1 patient were pyrexia (n = 4), constipation (n = 3), headache (n = 3) and peripheral edema (n = 2). Comparative analysis found no evidence of significant differences in zibotentan exposure between the Chinese patients in our study, and the previous Japanese and Caucasian studies. The PK and safety profiles of zibotentan determined in this Chinese patient population are similar to those previously reported. Our findings suggest no clinically relevant inter-ethnic differences in zibotentan disposition between the patient populations analyzed.


Relationship between asymmetry parameter and hemispheric backscatter ratio: implications for climate forcing by aerosols.
Calculations of direct climate forcing by anthropogenic aerosols commonly use radiative transfer parameters, including asymmetry parameter g. One method of obtaining the asymmetry parameter of a particle population is to convert measured values of the hemispheric-to-total-scatter ratio (backscatter ratio b) into their corresponding g values. We compare a conversion derived from Mie calculations with one derived from the Henyey-Greenstein (HG) phase function to show that the HG method systematically overestimates g for typical size distributions of accumulation-mode aerosols. A delta-Eddington radiative transfer calculation is used to show that a 10% overestimation of g can systematically reduce climate forcing as a result of aerosols by 12% or more. Mie computations are used to derive an empirical relationship between backscatter ratio and asymmetry parameter for log-normal accumulation-mode aerosols. This relationship can be used to convert the backscatter ratio to the asymmetry parameter, independent of geometric mean diameter D(gv) or complex refractive index m, but the conversion requires knowledge of the breadth σ(g) of the size distribution.


Meditation and neurodegenerative diseases.
Neurodegenerative diseases pose a significant problem for the healthcare system, doctors, and patients. With an aging population, more and more individuals are developing neurodegenerative diseases and there are few treatment options at the present time. Meditation techniques present an interesting potential adjuvant treatment for patients with neurodegenerative diseases and have the advantage of being inexpensive, and easy to teach and perform. There is increasing research evidence to support the application of meditation techniques to help improve cognition and memory in patients with neurodegenerative diseases. This review discusses the current data on meditation, memory, and attention, and the potential applications of meditation techniques in patients with neurodegenerative diseases.


Regulation of hepatic insulin receptors by pancreatic polypeptide in fasting and feeding.
Pancreatic polypeptide (PP) increases hepatic insulin receptor (IR) binding activity in fasted PP-deficient rats, but not fasted normal animals. PP-induced alteration of hepatic IR levels in normal animals may be detectable in the fed state when IR concentrations are lower than during fasting. In the current study, the effect of exogenous PP on IR concentrations in the fed and fasted states was determined in healthy 300- to 350-g male Sprague-Dawley rats. Ten animals were administered PP 100 microgram/kg/day for 3 days by intraperitoneal injection and 10 weight-matched control animals received saline vehicle. Five PP- and five saline-administered rats were fasted for 12 hr prior to organ procurement, while 5 PP- and 5 saline-treated rats were given free access to food for this period. Livers were removed and snap-frozen. IRs were isolated from solubilized hepatocyte membranes by affinity chromatography with agarose-bound wheat germ agglutinin. Western blots were performed using a specific antibody to the beta subunit of the IR, which was detected by a chemiluminescence technique after 45-min exposure to X-ray film. Exposed films were examined by scanning densitometry and IR concentration was expressed as absorbance units per milligram of hepatic protein (mean +/- SE). Statistical comparisons were by Student's t test with significance taken at P < 0.05. Feeding was associated with a significantly lower IR concentration in saline-administered animals compared with the fasted state (24.2 +/- 4.0 vs 53.3 +/- 11.1). PP administration in fed rats resulted in significantly increased IR concentration as compared with that seen in saline-administered fed animals (43.8 +/- 8.9 vs 24.2 +/- 4.0). This difference may be due to increased IR synthesis with long-term PP administration, and supports the role of PP as a regulatory factor in hepatic carbohydrate metabolism.


A synthetic peptide ELISA for the screening of rubella virus neutralizing antibodies in order to ascertain immunity.
One hundred and fifty-one human sera from patients exposed to rubella virus (RV) and shown to be negative for IgM antibodies were tested for total RV-IgG, hemagglutination inhibition (HAI) and for virus neutralizing (VN) antibodies using a peptide enzyme-linked immunosorbent assay (ELISA) based on BCH-178, a peptide representing one of several known neutralizing epitopes on RV hemagglutinin (E1). The data showed that, among 39 and 51 sera with HAI and RV-IgG titres of 1/128 and >150 IU/ml, respectively, neutralizing antibody readings using the BCH-178 ELISA were above cut-off values. However, 13% of HAI positive sera (titre > or =1/16) and 16% of RV-IgG ELISA positive sera (> or =20 IU/ml) were below the cut-off value of the BCH-178 ELISA. This may explain why several cases of congenital rubella syndrome (CRS) have been observed in spite of positive titres. We suggest that a diagnosis of sufficient immunity against RV infection or reinfection may be safer if an additional test detecting antibodies against VN RV epitopes is positive as well.


Physiotherapy intervention late after stroke and mobility.
To determine whether the intervention of a physiotherapist improved mobility in patients seen more than one year after stroke. Randomised crossover trial comparing two groups offered intervention by a physiotherapist, one immediately after entry into the trial and the other after a delay of three months. The intervention consisted of identifying problems and offering advice and help to solve the problems. Patients' homes in Oxfordshire. Patients who had reduced mobility due to a stroke more than one year before entry; 60 were recruited from a community stroke register and 34 in other ways. Standard measures of mobility including gait speed, functional ambulation categories, the Nottingham extended activities of daily living index, and individual items from the Barthel activities of daily living index and the Frenchay activities index. Measures of manual dexterity, depression, and anxiety were used as controls. 94 patients entered the trial and 49 were randomised to immediate and 45 to delayed physiotherapy; 89 were compared at the crossover point. At randomisation the two groups were comparable. At three months the group given early therapy showed an improvement in gait speed whereas the untreated group had declined (differences of -3.9 v 6.4 s to walk 10 m; p less than 0.01); between three and six months the group given delayed therapy showed improvement and the previously treated group declined (differences of 6.5 v -3.9 s to walk 10 m; p less than 0.01). A 9% (95% confidence interval 0% to 18%) decrease in time taken to walk 10 m was associated with treatment and a 12% (2% to 19%) increase when patients were untreated. Other measures did not change significantly. Intervention of an experienced physiotherapist late after stroke specifically improves mobility, albeit by a small amount, but the effects do not seem to be maintained, perhaps because there is an underlying decline in mobility in these patients. Gait speed offers a simple and sensitive measure of outcome.


Identification of novel argonaute-associated proteins.
RNA silencing processes are guided by small RNAs known as siRNAs and microRNAs (miRNAs) . They reside in ribonucleoprotein complexes, which guide the cleavage of complementary mRNAs or affect stability and translation of partial complementary mRNAs . Argonaute (Ago) proteins are at the heart of silencing effector complexes and bind the single-stranded siRNA and miRNA . Our biochemical analysis revealed that Ago2 is present in a pre-miRNA processing complex that is able to transfer the miRNA into a target-mRNA cleaving complex. To gain insight into the function and composition of RNA silencing complexes, we purified Ago1- and Ago2-containing complexes from human cells. Several known Ago1- and/or Ago2-associated proteins including Dicer were identified, but also two novel factors, the putative RNA helicase MOV10, and the RNA recognition motif (RRM)-containing protein TNRC6B/KIAA1093. The new proteins localize, similar to Ago proteins, to mRNA-degrading cytoplasmic P bodies, and they are functionally required to mediate miRNA-guided mRNA cleavage.


An epigenetic signature of developmental potential in neural stem cells and early neurons.
A cardinal property of neural stem cells (NSCs) is their ability to adopt multiple fates upon differentiation. The epigenome is widely seen as a read-out of cellular potential and a manifestation of this can be seen in embryonic stem cells (ESCs), where promoters of many lineage-specific regulators are marked by a bivalent epigenetic signature comprising trimethylation of both lysine 4 and lysine 27 of histone H3 (H3K4me3 and H3K27me3, respectively). Bivalency has subsequently emerged as a powerful epigenetic indicator of stem cell potential. Here, we have interrogated the epigenome during differentiation of ESC-derived NSCs to immature GABAergic interneurons. We show that developmental transitions are accompanied by loss of bivalency at many promoters in line with their increasing developmental restriction from pluripotent ESC through multipotent NSC to committed GABAergic interneuron. At the NSC stage, the promoters of genes encoding many transcriptional regulators required for differentiation of multiple neuronal subtypes and neural crest appear to be bivalent, consistent with the broad developmental potential of NSCs. Upon differentiation to GABAergic neurons, all non-GABAergic promoters resolve to H3K27me3 monovalency, whereas GABAergic promoters resolve to H3K4me3 monovalency or retain bivalency. Importantly, many of these epigenetic changes occur before any corresponding changes in gene expression. Intriguingly, another group of gene promoters gain bivalency as NSCs differentiate toward neurons, the majority of which are associated with functions connected with maturation and establishment and maintenance of connectivity. These data show that bivalency provides a dynamic epigenetic signature of developmental potential in both NSCs and in early neurons.


Background contamination by coplanar polychlorinated biphenyls (PCBs) in trace level high resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS) analytical procedures.
The addition of the "dioxin-like" polychlorinated biphenyl (PCB) congeners to the assessment of risk associated with the 2,3,7,8-chlorine substituted dioxins and furans has dramatically increased the number of laboratories worldwide that are developing analytical procedures for their detection and quantitation. Most of these procedures are based on established sample preparation and analytical techniques employing high resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS), which are used for the analyses of dioxin/furans at low parts-per-trillion (ppt) levels. A significant and widespread problem that arises when using these sample preparation procedures for the analysis of coplanar PCBs is the presence of background levels of these congeners. Industrial processes, urban incineration, leaking electrical transformers, hazardous waste accidents, and improper waste disposal practices have released appreciable quantities of PCBs into the environment. This contamination has resulted in the global distribution of these compounds via the atmosphere and their ubiquitous presence in ambient air. The background presence of these compounds in method blanks must be addressed when determining the exact concentrations of these and other congeners in environmental samples. In this study reliable procedures were developed to accurately define these background levels and assess their variability over the course of the study. The background subtraction procedures developed and employed increase the probability that the values reported accurately represent the concentrations found in the samples and were not biased due to this background contamination.


Ischemic vagus nuclei lesions and hyperglycemia: a study in 26 patients with lateral medullary infarction and matched controls.
Hyperglycemia is common after stroke in diabetic and nondiabetic patients. Furthermore, it has been associated with infarct expansion, worse functional outcomes and higher mortality. In a previous study, infarction of the insular region was related to higher poststroke glucose levels than infarcts in other cortical areas. Experimental studies in animal models suggested that the lower brainstem nuclei of the vagus nerve modulate insulin secretion. These nuclei are usually affected in lateral medullary infarction (LMI). We evaluated whether patients with lateral medullary stroke have worse poststroke glycemic control than other stroke patients. A hospital-based stroke registry was used to identify 26 patients from the years 2000 to 2010 who fulfilled the following inclusion criteria: (1) a first-ever stroke; (2) neurological deficits compatible with LMI; (3) MRI confirmation of an ischemic lesion of the lateral medulla involving the vagus nerve nuclei, and (4) no simultaneous infarcts. Patients were excluded if they were admitted to the hospital more than 24 h after stroke onset or died in the first 24 h after hospital admission. A control group of other stroke patients was randomly selected from the same stroke registry and over the same time period, matching for the age and gender of the LMI group. The average glycemia was compared between the two groups using a linear regression model adjusted for confounders. Glycated hemoglobin at admission was used to estimate prestroke glycemic control. Prestroke glycemic averages were then compared with poststroke glycemia for the two groups using the Wilcoxon signed test for related samples. The average glycemia of the LMI patients in the first 24 h after stroke was 9.4 mmol/l (SD 3.2), and from 24 to 72 h it was 7.6 mmol/l (SD 2.8). In the comparison group, these values were 7.7 (SD 2.8) and 7.1 mmol/l (SD 2.7), respectively. As expected, diabetic patients had a significantly higher glycemia than nondiabetic patients (p < 0.0001). The adjusted linear regression model showed the average glycemia differences to be significant for the first 24 h (p = 0.001; R(2) = 55.6%) but not for the 24- 72 h period. The frequency of previous diabetes mellitus was similar in both groups. As compared to prestroke glycemic estimates, glycemia in lateral medullary stroke patients increased significantly more than in controls during the first 24 h after stroke (p = 0.01), but again there were no significant differences for the 24-72 h period. This study suggests that ischemic lesions of the vagus nerve nuclei are associated with worse early poststroke glycemic control than stroke in other locations. Confirmation of this hypothesis and the long-term implications of glucose control impairment warrant further prospective studies.


Effects of calcium and vitamin D on MLH1 and MSH2 expression in rectal mucosa of sporadic colorectal adenoma patients.
To further clarify and develop calcium and vitamin D as chemopreventive agents against colorectal cancer in humans and develop modifiable biomarkers of risk for colorectal cancer, we conducted a pilot, randomized, double-blind, placebo-controlled, 2 x 2 factorial clinical trial to test the effects of calcium and vitamin D(3), alone and in combination, on key DNA mismatch repair proteins in the normal colorectal mucosa. Ninety-two men and women with at least one pathology-confirmed colorectal adenoma were treated with 2.0 g/d calcium or 800 IU/d vitamin D(3), alone or in combination, versus placebo over 6 months. Colorectal crypt overall expression and distribution of MSH2 and MLH1 proteins in biopsies of normal-appearing rectal mucosa were detected by automated immunohistochemistry and quantified by image analysis. After 6 months of treatment, MSH2 expression along the full lengths of crypts increased by 61% (P = 0.11) and 30% (P = 0.36) in the vitamin D and calcium groups, respectively, relative to the placebo group. The estimated calcium and vitamin D treatment effects were more pronounced in the upper 40% of crypts (differentiation zone) in which MSH2 expression increased by 169% (P = 0.04) and 107% (P = 0.13) in the vitamin D and calcium groups, respectively. These findings suggest that higher calcium and vitamin D intakes may result in increased DNA MMR system activity in the normal colorectal mucosa of sporadic adenoma patients and that the strongest effects may be vitamin D related and in the differentiation zone of the colorectal crypt.


Functional alterations of myeloid cells during the course of Alzheimer's disease.
Neuroinflammation is a hallmark of neurodegenerative disease and a significant component of the pathology of Alzheimer's disease (AD). Patients present with extensive microgliosis along with elevated pro-inflammatory signaling in the central nervous system and periphery. However, the role of peripheral myeloid cells in mediating and influencing AD pathogenesis remains unresolved. Peripheral myeloid cells were isolated from peripheral blood of patients with prodromal AD (n = 44), mild AD dementia (n = 25), moderate/severe AD dementia (n = 28), and age-matched controls (n = 54). Patients were evaluated in the clinic for AD severity and categorized using Clinical Dementia Rating (CDR) scale resulting in separation of patients into prodromal AD (CDR0.5) and advancing forms of AD dementia (mild-CDR1 and moderate/severe-CDR2/3). Separation of peripheral myeloid cells into mature monocytes or immature MDSCs permitted the delineation of population changes from flow cytometric analysis, RNA phenotype analysis, and functional studies using T cell suppression assays and monocyte suppression assays. During stages of AD dementia (CDR1 and 2/3) peripheral myeloid cells increase their pro-inflammatory gene expression while at early stages of disease (prodromal AD-CDR0.5) pro-inflammatory gene expression is decreased. MDSCs are increased in prodromal AD compared with controls (16.81% vs 9.53%) and have markedly increased suppressive functions: 42.4% suppression of activated monocyte-produced IL-6 and 78.16% suppression of T cell proliferation. In AD dementia, MDSC populations are reduced with decreased suppression of monocyte IL-6 (5.22%) and T cell proliferation (37.61%); the reduced suppression coincides with increased pro-inflammatory signaling in AD dementia monocytes. Peripheral monocyte gene expression is pro-inflammatory throughout the course of AD, except at the earliest, prodromal stages when pro-inflammatory gene expression is suppressed. This monocyte biphasic response is associated with increased numbers and suppressive functions of MDSCs during the early stages and decreased numbers and suppressive functions in later stages of disease. Prolonging the early protective suppression and reversing the later loss of suppressive activity may offer a novel therapeutic strategy.


Epigenetic inactivation of inositol polyphosphate 4-phosphatase B (INPP4B), a regulator of PI3K/AKT signaling pathway in EBV-associated nasopharyngeal carcinoma.
Nasopharyngeal carcinoma (NPC) is a common viral-associated neoplasm in which multiple signaling cascades are interfered with by Epstein-Bar virus (EBV) latent proteins and various genetic alterations. Aside from the previously reported PIK3CA amplification, we examined the role of INPP4B, a negative regulator of the PI3K/AKT signaling pathway in the development of NPC. By RT-PCR and Western blotting, we revealed that the expression of INPP4B was down-regulated in all five established EBV-positive tumor lines. While INPP4B was consistently expressed in normal nasopharyngeal epithelial cells, downregulation of INPP4B was found in 32/65 (49.2%) of primary tumors by immunohistochemistry. Furthermore, our study also demonstrated the hypermethylation of the 5'CpG island of INPP4B in the tumors in which INPP4B transcription was downregulated. Notably, the re-expression of INPP4B was detected in the NPC cells treated with the demethylation agent (5-aza-2'deoxycytidine). Our study showed that promoter hypermethylation was the major mechanism for transcriptional silencing of INPP4B in NPC. Furthermore, restoration of INPP4B expression significantly suppressed PI3K/AKT downstream signals in the NPC C666-1 cells. In vivo growth inhibition was clearly demonstrated in the tumor cells stably expressing INPP4B. The findings indicate that epigenetic inactivation of INPP4B is one of the key mechanisms in activating PI3K/AKT signaling cascade and playing a role in the tumorigenesis of NPC.


Increased arachidonate in lipids after administration to man: effects on prostaglandin biosynthesis.
Ethyl arachidonate was administered orally to 4 healthy male volunteers in a dose of 6 gm daily for a 2 to 3 wk period after 10-day control period. The increased intake of this precursor of the dienoic prostaglandins resulted in significant increases in the relative and absolute amount of arachidonate in plasma triglycerides, phospholipids, and cholesteryl esters. Similar changes in lipid composition were noted in platelets. The excretion of 7alpha-hydroxy-5,11-diketotetranoprostane-1,16-dioic acid, the major urinary metabolite of E prostaglandins in man, was increased by an average of 47% in 3 of the 4 volunteers. Platelet reactivity was assessed by determining the threshold concentration of adenosine diphosphate (ADP) necessary to induce secondary, irreversible aggregation of platelet-rich plasma. This threshold concentration dropped significantly in all volunteers (10% to 60% of control values). It is concluded that the biosynthesis and function of prostaglandins can be augmented in man by oral administration of an esterified precursor fatty acid.


QCScreen: a software tool for data quality control in LC-HRMS based metabolomics.
Metabolomics experiments often comprise large numbers of biological samples resulting in huge amounts of data. This data needs to be inspected for plausibility before data evaluation to detect putative sources of error e.g. retention time or mass accuracy shifts. Especially in liquid chromatography-high resolution mass spectrometry (LC-HRMS) based metabolomics research, proper quality control checks (e.g. for precision, signal drifts or offsets) are crucial prerequisites to achieve reliable and comparable results within and across experimental measurement sequences. Software tools can support this process. The software tool QCScreen was developed to offer a quick and easy data quality check of LC-HRMS derived data. It allows a flexible investigation and comparison of basic quality-related parameters within user-defined target features and the possibility to automatically evaluate multiple sample types within or across different measurement sequences in a short time. It offers a user-friendly interface that allows an easy selection of processing steps and parameter settings. The generated results include a coloured overview plot of data quality across all analysed samples and targets and, in addition, detailed illustrations of the stability and precision of the chromatographic separation, the mass accuracy and the detector sensitivity. The use of QCScreen is demonstrated with experimental data from metabolomics experiments using selected standard compounds in pure solvent. The application of the software identified problematic features, samples and analytical parameters and suggested which data files or compounds required closer manual inspection. QCScreen is an open source software tool which provides a useful basis for assessing the suitability of LC-HRMS data prior to time consuming, detailed data processing and subsequent statistical analysis. It accepts the generic mzXML format and thus can be used with many different LC-HRMS platforms to process both multiple quality control sample types as well as experimental samples in one or more measurement sequences.


[Establishment of an optimum bowel preparation method before gynecologic laparoscopic surgery].
To establish a new method for preoperative bowel preparation that facilitates nursing care and minimizes the patient's discomfort during the clinical pathway of laparoscopic surgery. A randomized controlled trial was conducted for the following two preparation methods. Twenty cases were assessed with Method 1 and 18 cases with Method 2. Method 1 (the conventional procedure): oral magnesium citrate is given in the afternoon of the day before surgery, followed by a glycerin enema in the night of the day before surgery and in the morning of the day of surgery. Method 2 (a new procedure): oral magnesium citrate is given in the afternoon of the day before surgery, followed by oral picosulfate in the night before the day of surgery and a bisacodyl suppository in the morning of the day of surgery. To evaluate the two methods we sent questionnaires to the surgeons (blinded to the method used), nurses, and patients. No statistical difference existed between the two methods in their effectiveness as a preoperative treatment. Facilitation of nursing care was significantly better in Method 2, and patients had considerably reduced discomfort with Method 2. Patients who received oral picosulfate and a bisacodyl suppository experienced much less discomfort and nursing care was easier when compared with the conventional method of administering a glycerin enema. Since an enema is disliked by young women and an effect comes out with discomfort very shortly after the administration, the degree of discomfort of patients would have become high. Picosulfate is an oral medicine and thereby the effect comes out mildly. That would be the reason why the degree of discomfort of patients was low. In the nursing care, an enema requires time for preparation and administration, while picosulfate is easy to administer, making the nursing care easier. Therefore, Method 2 was chosen as a preoperative bowel treatment for the clinical pathway. Thus, we could establish a new evidence-based method useful for the preoperative bowel preparation in the clinical pathway of laparoscopic surgery.


Improvements in early mortality and coagulopathy are sustained better in patients with blunt trauma after institution of a massive transfusion protocol in a civilian level I trauma center.
Transfusion practices across the country are changing with aggressive use of plasma (fresh-frozen plasma [FFP]) and platelets during massive transfusion with current military recommendations to use component therapy at a 1:1:1 ratio of packed red blood cells to FFP to platelets. A massive transfusion protocol (MTP) was designed to achieve a packed red blood cell:FFP:platelet ratio of 1:1:1 We prospectively gathered demographic, transfusion, and patient outcome data during the first year of the MTP and compared this with a similar cohort of injured patients (pre-MTP) receiving > or = 10 red blood cell (RBC) in the first 24 hours of hospitalization before instituting the MTP. One hundred sixteen MTP activations occurred. Twelve non-trauma patients and 31 who did not receive 10 RBC (15 deaths, 16 early bleeding controls) were excluded. Seventy-three MTP patients were compared with 84 patients with pre-MTP who had similar demographics and injury severity score (29 vs. 29, p = 0.99). MTP patients received an average of 23.7 RBC and 15.6 FFP transfusions compared with 22.8 RBC (p = 0.67) and 7.6 FFP (p < 0.001) transfusions in pre-MTP patients. Early crystalloid usage dropped from 9.4 L (pre-MTP) to 6.9 L (MTP) (p = 0.006). Overall patient mortality was markedly improved at 24 hours, from 36% in the pre-MTP group to 17% in the MTP group (p = 0.008) and at 30 days (34% mortality MTP group vs. 55% mortality in pre-MTP group, p = 0.04). Blunt trauma survival improvements were more marked and more sustained than victims of penetrating trauma. Early deaths from coagulopathic bleeding occurred in 4 of 13 patients in the MTP group vs. 21 of 31 patients in the pre-MTP group (p = 0.023). In the civilian setting, aggressive use of FFP and platelets drastically reduces 24-hour mortality and early coagulopathy in patients with trauma. Reduction in 30 day mortality was only seen after blunt trauma in this small subset.


Feasibility and validity of telephone triage for adverse events during a voluntary medical male circumcision campaign in Swaziland.
Voluntary medical male circumcision (VMMC) reduces HIV acquisition among heterosexual men by approximately 60%. VMMC is a surgical procedure and some adverse events (AEs) are expected. Swaziland's Ministry of Health established a toll-free hotline to provide general information about VMMC and to manage post-operative clinical AEs through telephone triage. We retrospectively analyzed a dataset of telephone calls logged by the VMMC hotline during a VMMC campaign. The objectives were to determine reasons clients called the VMMC hotline and to ascertain the accuracy of telephone-based triage for VMMC AEs. We then analyzed VMMC service delivery data that included date of surgery, AE type and severity, as diagnosed by a VMMC clinician as part of routine post-operative follow-up. Both datasets were de-identified and did not contain any personal identifiers. Proportions of AEs were calculated from the call data and from VMMC service delivery data recorded by health facilities. Sensitivity analyses were performed to assess the accuracy of phone-based triage compared to clinically confirmed AEs. A total of 17,059 calls were registered by the triage nurses from April to December 2011. Calls requesting VMMC education and counseling totaled 12,492 (73.2%) and were most common. Triage nurses diagnosed 384 clients with 420 (2.5%) AEs. According to the predefined clinical algorithms, all moderate and severe AEs (153) diagnosed through telephone-triage were referred for clinical management at a health facility. Clinicians at the VMMC sites diagnosed 341 (4.1%) total clients as having a mild (46.0%), moderate (47.8%), or severe (6.2%) AE. Eighty-nine (26%) of the 341 clients who were diagnosed with AEs by clinicians at a VMMC site had initially called the VMMC hotline. The telephone-based triage system had a sensitivity of 69%, a positive predictive value of 83%, and a negative predictive value of 48% for screening moderate or severe AEs of all the AEs. The use of a telephone-based triage system may be an appropriate first step to identify life-threatening and urgent complications following VMMC surgery.


Effect of NGF treatment on outcome measures in a rat model of middle cerebral artery occlusion.
Ischemic insults to the brain result in a time-dependent increase in neuronal death that is responsible for some of the functional deficits associated with stroke. Our working hypothesis is that ischemia results in a prompt depletion of high energy phosphate species resulting in decreased pH and glutathione levels in brain in a temporal and spatial pattern that disrupts nerve growth factor homeostasis and increases neuronal apoptosis. Here we show hemispheric depletion of active phosphate species after ischemia. Also, we observed that the striatum is an early target for oxidative stress that is followed by energy metabolic impairment and altered neurotrophin levels that were detected by noninvasive magnetic resonance imaging (MRI) measurements of cytotoxicity and conventional biochemical determinations of apoptosis, glutathione, and nerve growth factor (NGF) protein levels in a pattern distinct from that observed in the hippocampus. Furthermore, early assessment of intracellular pH by 31P-magnetic resonance spectroscopy (31P-MRS) was a predictor of later infarct development as determined by MRI. We also show that pretreatment with pharmacological doses of NGF did not have overall significant beneficial consequences on irreversible ischemia in an intraluminal unilateral irreversible model of stroke in rat brain.


How can we improve crop genotypes to increase stress resilience and productivity in a future climate? A new crop screening method based on productivity and resistance to abiotic stress.
The need to accelerate the selection of crop genotypes that are both resistant to and productive under abiotic stress is enhanced by global warming and the increase in demand for food by a growing world population. In this paper, we propose a new method for evaluation of wheat genotypes in terms of their resilience to stress and their production capacity. The method quantifies the components of a new index related to yield under abiotic stress based on previously developed stress indices, namely the stress susceptibility index, the stress tolerance index, the mean production index, the geometric mean production index, and the tolerance index, which were created originally to evaluate drought adaptation. The method, based on a scoring scale, offers simple and easy visualization and identification of resilient, productive and/or contrasting genotypes according to grain yield. This new selection method could help breeders and researchers by defining clear and strong criteria to identify genotypes with high resilience and high productivity and provide a clear visualization of contrasts in terms of grain yield production under stress. It is also expected that this methodology will reduce the time required for first selection and the number of first-selected genotypes for further evaluation by breeders and provide a basis for appropriate comparisons of genotypes that would help reveal the biology behind high stress productivity of crops.


Electromagnetic shielding effectiveness of thin film with composite carbon nanotubes and stainless steel fibers.
Using the polymer blending method, conductive materials and waterborne polyurethane (WPU) were mixed to fabricate conductive composite films for application in electromagnetic shielding. First, nitric acid was used to purify the multi-walled carbon nanotubes (MWCNT). Second, sodium dodecyl sulfate (SDS) was utilized to disperse the carbon nanotubes, and then they were mixed with 8 microm diameter and 2 mm long stainless steel fibers (SSF) in the WPU by the polymer blending method. Finally, the thickness of 0.25 mm of conductive composite film was fabricated by means of coating. According to the ASTM D4935-99 standard, a coaxial transmission line was used to measure the electromagnetic shielding effectiveness (EMSE) of conductive composite film within the range of 50 MHz approximately 3.0 GHz. Moreover, the influence of the prior and posterior dispersion of carbon nanotubes dispersed on electromagnetic shielding was dealt with in the paper. Results demonstrated that the conductive composite film, within 50 MHz approximately 3.0 GHz, fabricated by the 15 wt% of the multi-walled carbon nanotubes and 30 wt% of the stainless steel fibers can achieve the maximum of the electromagnetic shielding effectiveness, 34.86 dB, and its shielding effect, 99.9%.


Self-organization of bidisperse colloids in water droplets.
Most of the colloidal clusters have been produced from oil-in-water emulsions with identical microspheres dispersed in oil droplets. Here, we present new types of binary colloidal clusters from phase-inverted water-in-oil emulsions using various combinations of two different colloids with several size ratios: monodisperse silica or polystyrene microspheres for larger particles and silica or titania nanoparticles for smaller particles. Obviously, a better understanding of how finite groups of different colloids self-organize in a confined geometry may help us control the structure of matter at multiple length scales. In addition, since aqueous dispersions have much better phase stability, we could produce much more diverse colloidal materials from water-in-oil emulsions rather than from oil-in-water emulsions. Interestingly, the configurations of the large microspheres were not changed by the presence of the small particles. However, the arrangement of the smaller particles was strongly dependent on the nature of the interparticle interactions. The experimentally observed structural evolutions were consistent with the numerical simulations calculated using Surface Evolver. These clusters with nonisotropic structures can be used as building blocks for novel colloidal structures with unusual properties or by themselves as light scatterers, diffusers, and complex adaptive matter exhibiting emergent behavior.


Corneal Deformation Response in Patients With Primary Open-Angle Glaucoma and in Healthy Subjects Analyzed by Corvis ST.
To compare corneal deformation response between patients with primary open-angle glaucoma (POAG) and healthy subjects. A prevalent case-control study was conducted, followed by an integrated meta-analysis. Primary open-angle glaucoma was confirmed by presence of glaucomatous optic disc damage with corresponding visual field defect after excluding secondary causes. Age-matched controls were recruited. Corvis ST (CST) was used to measure 10 parameters of corneal deformation response and central corneal thickness (CCT). The association between parameters of deformation response and clinical factors was assessed by the linear regression analyses. Differences in deformation response between POAG and healthy subjects were compared after adjusting for other factors. Eligible studies were identified by a systematic search of the PubMed, ISI Web of Science, and Embase databases; Web sites of professional associations; and the Google Scholar engine. This study included 37 patients with POAG and 36 healthy controls. Significant associations were found between IOP and the first applanation time (A1T), first applanation velocity (A1V), second applanation time (A2T), and second applanation velocity (A2V) in each group. The deformation amplitude (DA), A1V, and A2T were lower in the POAG group, whereas the A1T, A2V and peak distance (PD) were greater in the POAG group than in the healthy controls (all P < 0.05). Ten separate studies plus the present study, involving 691 patients with POAG and 740 controls, were ultimately meta-analyzed. The significant differences in the DA, A1T, and A2T were further confirmed, with pooled weighted mean difference (WMD) of 0.13 mm (95% CI 0.04-0.23; P = 0.008) for A1T, -0.13 mm (95% CI -0.21 to -0.05; P = 0.001) for DA, and -0.46 ms (95% CI -0.70 to -0.22; P < 0.001) for A2T. The corneal response parameters provided by CST are informative for the assessment of corneal biomechanics. Patients with POAG showed significantly greater A1T and lower DA and A2T values than healthy controls, indicating a less deformable cornea in POAG patients.


PLANT DIVERSITY OF WESTERN CHITWAN FLORISTIC APPROACH.
This paper identifies and documents the plant biodiversity of western Chitwan, Nepal. Specifically, our attention was focussed on the plants of forests, grasslands and common lands based on our "Reciprocal Relation of Population and Environment Study Project" conducted during January-April 1996. This species-diversity of trees, shrubs and herbaceous flora was recorded from 117, 117, and 1049 sampling quadrats of 10×10, 3×3 and 1×1 m(2), respectively. The flora of our study plots contains 236 species that belong to 191 genera and 66 families. Of 236 species of plants, 119, 113, 59, 35 and 119 species were recorded from Tikauli forest, National Park forest, forests along the Narayani river, grasslands of National Park and common lands of the western Chitwan, respectively. Dicotyledons represent 184 (77.97%) species of the total flora species followed by monocotyledons (46 spp., 19.49%) and ferns (6 spp., 2.54%), respectively. The five largest families are Leguminosae (38 spp.), Poaceae (27 spp.), Asteraceae (22 spp.), Rubiaceae (10 spp.), and Scrophulariaceae (9 spp.). Hedyotis, Grewia and Lindernia, each with 4 spp., are the most speciose genera in the flora.


Identification and expression of NEU3, a novel human sialidase associated to the plasma membrane.
Several mammalian sialidases have been described so far, suggesting the existence of numerous polypeptides with different tissue distributions, subcellular localizations and substrate specificities. Among these enzymes, plasma-membrane-associated sialidase(s) have a pivotal role in modulating the ganglioside content of the lipid bilayer, suggesting their involvement in the complex mechanisms governing cell-surface biological functions. Here we describe the identification and expression of a human plasma-membrane-associated sialidase, NEU3, isolated starting from an expressed sequence tag (EST) clone. The cDNA for this sialidase encodes a 428-residue protein containing a putative transmembrane helix, a YRIP (single-letter amino acid codes) motif and three Asp boxes characteristic of sialidases. The polypeptide shows high sequence identity (78%) with the membrane-associated sialidase recently purified and cloned from Bos taurus. Northern blot analysis showed a wide pattern of expression of the gene, in both adult and fetal human tissues. Transient expression in COS7 cells permitted the detection of a sialidase activity with high activity towards ganglioside substrates at a pH optimum of 3.8. Immunofluorescence staining of the transfected COS7 cells demonstrated the protein's localization in the plasma membrane.