Supramolecular architecture based on the self-assembling of multiwall carbon nanotubes dispersed in polyhistidine and glucose oxidase: Characterization and analytical applications for glucose biosensing.
We report for the first time the development of a sensitive and selective glucose biosensor based on the self-assembling of multiwall carbon nanotubes (MWCNTs) dispersed in polyhistidine (Polyhis) and glucose oxidase (GOx) on glassy carbon electrodes (GCE). The supramolecular architecture was characterized by SEM, FT-IR and electrochemical techniques. The optimum multistructure was obtained with five (MWCNT-Polyhis/GOx) bilayers and one layer of Nafion as anti-interferent barrier. The sensitivity at 0.700V was (1.94±0.03) mAM(-1) (r=0.9991), with a linear range between 0.25 and 5.00mM, a detection limit of 2.2μM and a quantification limit of 6.7μM with minimum interference from lactose (1.5%), maltose (5.7%), galactose (1.2%), ascorbic acid (1.0%), and uric acid (3.3%). The biocatalytic layer demonstrated to be highly reproducible since the R.S.D. for 10 successive amperometric calibrations using the same surface was 3.6%. The sensitivity of the biosensor after 15 day storage at 4°C remained at 90% of its original value. The combination of the excellent dispersing properties and polycationic nature of polyhistidine, the stability of the MWCNT-Polyhis dispersion, the electrocatalytic properties of MWCNTs, the biocatalytic specificity of GOx, and the permselective properties of Nafion have allowed building up a sensitive, selective, robust, reproducible and stable glucose amperometric biosensor for the quantification of glucose in milk samples.


I405V polymorphism of CETP gene and lipid profile in women with endometriosis.
Genetic factors have an important role in the pathophysiology of endometriosis. In addition, abnormalities in lipid profile and intrinsic inflammatory status are associated with disease progression. The purpose of this study was to evaluate the effect of the I405V polymorphism of cholesteryl ester transfer protein (CETP) gene and lipid profile with the risk of endometriosis in women. Ninety-seven women with laparoscopy-diagnosed endometriosis were recruited for this study, and 107 patients with no evidence of endometriosis confirmed by laparoscopy served as controls. Samples were analyzed for polymorphism of the CETP gene using polymerase chain reaction-restriction fragment length polymorphism-based methods. After adjustment for body mass index, high-density lipoprotein-C and low-density lipoprotein-C, the risk of endometriosis in patients with normal genotype homozygous was more of the rare allele (p < 0.001, odds ratio = 0.21, 95% confidence interval = 0.09-0.45). Our results suggest that I405V polymorphism of CETP gene plays an important role as independent factor in the risk of endometriosis in women.


Effect of oestradiol and insulin on the proliferative pattern and on oestrogen and progesterone receptor contents in MCF-7 cells.
In a long-term experiment MCF-7 cells showed an exponential proliferation rate in fetal calf serum while they become quiescent in growth-factor- and steroid-free serum. The mitogenic activity of this cell line was increased by oestradiol and insulin, exposure to both hormones showing a synergic effect. These mitogen-mediated effects are inhibited by genistein a phytoestrogen which, by itself at the doses used, did not affect either the basal proliferation rate or the total protein content. Immunoblotting and Scatchard analysis reveal respectively that insulin increases the oestrogen receptor (ER) content and its binding capacity. The presence of genistein decreases the ER content and completely abolishes the binding capacity of this protein. Insulin is able to increase progesterone receptor (PR) levels while, in the presence of genistein, the above-reported effect was completely abolished. On the basis of the latter data the authors suggest that insulin is able to affect the phosphorylation status of ER and PR, inducing functional changes in both proteins, although this was in the absence of their natural ligands. Indeed, the presence in the medium of a tyrosine kinase inhibitor such as genistein could substantially decrease both ER and PR levels in these cells.


Effect of perinatal exposure to methadone on brain opioid and alpha 2-adrenergic receptors.
Prenatal exposure to psychotropic drugs has recently been shown to induce behavioral and brain neurochemical changes in the newborn. The long-term effects of pre- and postnatal exposure to methadone on the opioid (delta and mu) and adrenergic (alpha 2) receptors were investigated in two areas of the rat brain. Rat pups received prenatal treatment with methadone through a minipump implanted in the pregnant dams and were cross-fostered to methadone- or non-methadone-implanted dams until weaned. The rats were sacrificed when fully grown and brains removed. Receptor binding assays using 3H-D-ala-leu-enkephalin and 3H-naloxone for delta and mu opioid receptors respectively and 3H-rauwolscine for alpha 2-adrenergic receptors were performed in two areas of the brain. In the hypothalamus, prenatal methadone treatment induced sustained decreases in both delta and mu opioid receptors but did not cause significant changes in the alpha 2-adrenergic receptors. In the cerebral cortex, prenatal methadone treatment induced significant decreases not only in delta and mu opioid receptors but also in alpha 2-adrenergic receptors. Postnatal methadone treatment alone did not cause significant changes in opioid receptors in the two brain areas, but did induce reduction in the cerebral cortical alpha 2-adrenergic receptors. The significance of these results is discussed.


Sequential inverse dysregulation of the RNA helicases DDX3X and DDX3Y facilitates MYC-driven lymphomagenesis.
DDX3X is a ubiquitously expressed RNA helicase involved in multiple stages of RNA biogenesis. DDX3X is frequently mutated in Burkitt lymphoma, but the functional basis for this is unknown. Here, we show that loss-of-function DDX3X mutations are also enriched in MYC-translocated diffuse large B cell lymphoma and reveal functional cooperation between mutant DDX3X and MYC. DDX3X promotes the translation of mRNA encoding components of the core translational machinery, thereby driving global protein synthesis. Loss-of-function DDX3X mutations moderate MYC-driven global protein synthesis, thereby buffering MYC-induced proteotoxic stress during early lymphomagenesis. Established lymphoma cells restore full protein synthetic capacity by aberrant expression of DDX3Y, a Y chromosome homolog, the expression of which is normally restricted to the testis. These findings show that DDX3X loss of function can buffer MYC-driven proteotoxic stress and highlight the capacity of male B cell lymphomas to then compensate for this loss by ectopic DDX3Y expression.


Structural characterization of the interaction of α-synuclein nascent chains with the ribosomal surface and trigger factor.
The ribosome is increasingly becoming recognized as a key hub for integrating quality control processes associated with protein biosynthesis and cotranslational folding (CTF). The molecular mechanisms by which these processes take place, however, remain largely unknown, in particular in the case of intrinsically disordered proteins (IDPs). To address this question, we studied at a residue-specific level the structure and dynamics of ribosome-nascent chain complexes (RNCs) of α-synuclein (αSyn), an IDP associated with Parkinson's disease (PD). Using solution-state nuclear magnetic resonance (NMR) spectroscopy and coarse-grained molecular dynamics (MD) simulations, we find that, although the nascent chain (NC) has a highly disordered conformation, its N-terminal region shows resonance broadening consistent with interactions involving specific regions of the ribosome surface. We also investigated the effects of the ribosome-associated molecular chaperone trigger factor (TF) on αSyn structure and dynamics using resonance broadening to define a footprint of the TF-RNC interactions. We have used these data to construct structural models that suggest specific ways by which emerging NCs can interact with the biosynthesis and quality control machinery.


[Chronic pulmonary hypertension of vascular origin, plexogenic pulmonary arteriopathy and the appetite depressant aminorex: addenda to an epidemic].
An epidemic of chronic pulmonary hypertension occurred in Austria, the Federal Republic of Germany, and Switzerland, starting in 1967, peaking in 1968/69, and disappearing after 1972. The mechanism leading to pulmonary hypertension was precapillary vascular obstruction due to plexogenic pulmonary arteriopathy. There was a close geographic and temporal relationship between the epidemic and the marketing and intake of the appetite-depressing drug aminorex fumarate (Menocil). The epidemic was limited to the three above countries where aminorex had been on sale. In the individual patient the symptoms, usually dyspnea, angina pectoris and syncope on exertion, used to follow the beginning of the drug in-take after one year. A similar phase shift could be observed between marketing of the anoretic and the incidence of patients with chronic pulmonary hypertension of vascular origin. The new disease is compared with known forms of pulmonary vascular obstruction. It cannot be distinguished from classical primary pulmonary hypertension or from recurrent silent pulmonary thromboembolism on either clinical or functional grounds; it has plexogenic pulmonary arteriography in common with the former. The prognosis, however, is different: survival is considerably longer in patients with aminorex-associated pulmonary hypertension, and a marked decrease in the pulmonary vascular obstruction after 10 years is no exception. Considering the closeness of the various associations between the event (i.e. the epidemic) and its suspected cause (the anoretic aminorex) from the viewpoint of epidemiological, pharmacological, morphological and prognostic findings and considerations, there is little doubt that aminorex, besides other partly known and partly unknown factors, can in fact favour or cause the development of plexogenic pulmonary arteriography and pulmonary hypertension.(ABSTRACT TRUNCATED AT 250 WORDS)


Epidermal growth factor receptor plays a role in the regulation of liver and plasma lipid levels in adult male mice.
Dsk5 mice have a gain of function in the epidermal growth factor receptor (EGFR), caused by a point mutation in the kinase domain. We analyzed the effect of this mutation on liver size, histology, and composition. We found that the livers of 12-wk-old male Dsk5 heterozygotes (+/Dsk5) were 62% heavier compared with those of wild-type controls (+/+). The livers of the +/Dsk5 mice compared with +/+ mice had larger hepatocytes with prominent, polyploid nuclei and showed modestly increased cell proliferation indices in both hepatocytes and nonparenchymal cells. An analysis of total protein, DNA, and RNA (expressed relative to liver weight) revealed no differences between the mutant and wild-type mice. However, the livers of the +/Dsk5 mice had more cholesterol but less phospholipid and fatty acid. Circulating cholesterol levels were twice as high in adult male +/Dsk5 mice but not in postweaned young male or female mice. The elevated total plasma cholesterol resulted mainly from an increase in low-density lipoprotein (LDL). The +/Dsk5 adult mouse liver expressed markedly reduced protein levels of LDL receptor, no change in proprotein convertase subtilisin/kexin type 9, and a markedly increased fatty acid synthase and 3-hydroxy-3-methyl-glutaryl-CoA reductase. Increased expression of transcription factors associated with enhanced cholesterol synthesis was also observed. Together, these findings suggest that the EGFR may play a regulatory role in hepatocyte proliferation and lipid metabolism in adult male mice, explaining why elevated levels of EGF or EGF-like peptides have been positively correlated to increased cholesterol levels in human studies.


Genetic relatedness, antimicrobial and biocide susceptibility comparative analysis of methicillin-resistant and -susceptible Staphylococcus pseudintermedius from Portugal.
Forty methicillin-resistant and -susceptible Staphylococcus pseudintermedius (MRSP and MSSP, respectively) from colonization and infection in dogs and cats were characterized for clonality, antimicrobial, and biocide susceptibility. MSSP were genetically more diverse than MRSP by multi-locus sequence typing and pulsed-field gel electrophoresis. Three different spa types (t06, t02, t05) and two SCCmec types (II-III and V) were detected in the MRSP isolates. All MRSP and two MSSP strains were multidrug-resistant. Several antibiotic resistance genes (mecA, blaZ, tet(M), tet(K), aac(6')-Ie-aph(2')-Ia, aph(3')-III, ant(6)-Ia, sat4, erm(B), lnu(A), dfr(G), and catp(C221)) were identified by microarray and double mutations in the gyrA and grlA genes and a single mutation in the rpoB gene were detected by sequence analysis. No differences were detected between MSSP and MRSP in the chlorhexidine acetate (CHA) minimum inhibitory concentrations (MICs). However, two MSSP had elevated MIC to triclosan (TCL) and one to benzalkonium chloride and ethidium bromide. One MSSP isolate harboured a qacA gene, while in another a qacB gene was detected. None of the isolates harboured the sh-fabI gene. Three of the biocide products studied had high bactericidal activity (Otodine(®), Clorexyderm Spot Gel(®), Dermocanis Piocure-M(®)), while Skingel(®) failed to achieve a five log reduction in the bacterial counting. S. pseudintermedius have become a serious therapeutic challenge in particular if methicillin- resistance and/or multidrug-resistance are involved. Biocides, like CHA and TCL, seem to be clinically effective and safe topical therapeutic options.


Severe aortic thrombosis in the neonate--successful treatment with low-molecular-weight heparin: two case reports and review of the literature.
Small- to moderate-sized aortic thrombi in the sick newborn are frequently demonstrated, however, severe aortic thrombosis is rarely encountered. We report two newborn infants presenting with signs of decreased lower limb perfusion shortly after birth. An occlusive infrarenal aortic thrombus was demonstrated in both infants by ultrasound. No predisposing condition for thrombosis could be found in the first infant; in the second, an umbilical arterial catheter was the likely cause. The infants were successfully treated with low-molecular-weight heparin. In both infants resolution of the thrombus was observed by ultrasound. No complications of treatment were encountered. The clinical spectrum of neonatal aortic thrombosis and treatment options are reviewed.


Tumor necrosis factor-alpha-converting enzyme (TACE) levels in periodontal diseases.
Tumor necrosis factor-alpha-converting enzyme (TACE) is a metalloprotease which can shed several cytokines from the cell membrane, including receptor activator of NF-kappaB ligand (RANKL). This study aimed to investigate the hypothesis that TACE would be elevated in the gingival crevicular fluid (GCF) of persons with periodontitis. Total TACE amounts in GCF were higher in persons with chronic and aggressive periodontitis than in those with gingivitis or in healthy persons. TACE concentrations in GCF were higher in persons with chronic and aggressive periodontitis than in those with gingivitis, although not significantly higher than in healthy persons. Persons with chronic periodontitis receiving immunosuppressive treatment exhibited over 10-fold lower TACE levels than the other periodontitis groups. TACE was positively correlated with probing pocket depth, clinical attachment levels, and RANKL concentrations in GCF. In conclusion, the increased GCF TACE levels in persons with periodontitis and their positive correlation with RANKL may indicate an association of this enzyme with alveolar bone loss, and may warrant special attention in future therapeutic approaches.


p53 represses c-Myc through induction of the tumor suppressor miR-145.
The tumor suppressor p53 negatively regulates a number of genes, including the proto-oncogene c-Myc, in addition to activating many other genes. One mechanism of the p53-mediated c-Myc repression may involve transcriptional regulation. However, it is not clear whether microRNAs (miRNAs) play a role in the p53-mediated posttranscriptional regulation of c-Myc. In this study, we show that a putative tumor suppressor, miR-145, is expressed through the phosphoinositide-3 kinase (PI-3K)/Akt and p53 pathways. Importantly, p53 transcriptionally induces the expression of miR-145 by interacting with a potential p53 response element (p53RE) in the miR-145 promoter. We further show that c-Myc is a direct target for miR-145. Although miR-145 silences the expression of c-Myc, anti-miR-145 enhances its expression. This specific silencing of c-Myc by miR-145 accounts at least in part for the miR-145-mediated inhibition of tumor cell growth both in vitro and in vivo. Finally, the blockade of miR-145 by anti-miR-145 is able to reverse the p53-mediated c-Myc repression. Together, these results define the role of miR-145 in the posttranscriptional regulation of c-Myc by p53 and suggest that, as a new member of the p53 regulatory network, miR-145 provides a direct link between p53 and c-Myc in this gene regulatory network.


1980 Volvo award in basic science. Proteoglycans in experimental intervertebral disc degeneration.
An animal model of intervertebral disc degeneration induced surgically by ventral nuclear herniation in the rabbit produces morphologic changes of disc degeneration. Histologic characteristics and proteoglycan changes have been studied at various times after herniation. After injury, there was metaplasia into fibrocartilage originating from the cells along the margins of the annular wound, with proliferation of cells changing almost the entire disc space into fibrocartilage. A vertebral osteophyte occurred through an endochondral ossification sequence. Aggregating proteoglycans had two periods of repletion in the early course of degeneration. The water content of the disc was rapidly but only transiently restored in the first two days after herniation, whilst the changes in the total proteoglycan content of the disc paralleled these changes. Hyaluronic acid content decreased rapidly after herniation, but the size of the proteoglycan monomers did not change with degeneration. It is suggested that loss of confined fluid mechanics signals an abortive repair attempt rather than that of biochemical changes in proteoglycans initiate disc degeneration.


Predictability of the future development of aggressive behavior of cranial dural arteriovenous fistulas based on decision tree analysis.
The severity of clinical signs and symptoms of cranial dural arteriovenous fistulas (DAVFs) are well correlated with their pattern of venous drainage. Although the presence of cortical venous drainage can be considered a potential predictor of aggressive DAVF behaviors, such as intracranial hemorrhage or progressive neurological deficits due to venous congestion, accurate statistical analyses are currently not available. Using a decision tree data mining method, the authors aimed at clarifying the predictability of the future development of aggressive behaviors of DAVF and at identifying the main causative factors. Of 266 DAVF patients, 89 were eligible for analysis. Under observational management, 51 patients presented with intracranial hemorrhage/infarction during the follow-up period. The authors created a decision tree able to assess the risk for the development of aggressive DAVF behavior. Evaluated by 10-fold cross-validation, the decision tree's accuracy, sensitivity, and specificity were 85.28%, 88.33%, and 80.83%, respectively. The tree shows that the main factor in symptomatic patients was the presence of cortical venous drainage. In its absence, the lesion location determined the risk of a DAVF developing aggressive behavior. Decision tree analysis accurately predicts the future development of aggressive DAVF behavior.


Respiratory gene expression in soybean cotyledons during post-germinative development.
Gene expression for nuclear- and mitochondrial-encoded subunits of respiratory chain components was measured in developing soybean cotyledons and compared to the abundance of the relevant proteins. Overall respiratory gene expression peaked at day 16, close to the peak in cytochrome chain and TCA cycle activities from day 10 to 15. Protein abundance followed transcript abundance for all components examined with the exception of the F1beta subunit of ATP synthase. A dramatic peak in F1beta transcript levels early in development (day 5 to 7) was not mirrored by an increase in protein suggesting translational or post-translational control. Mitochondrial-encoded transcripts were at least 10-fold more abundant than nuclear-encoded transcripts. The pattern of transcript and protein abundance for uncoupling proteins displayed a trend similar to other respiratory proteins examined, implicating similar control mechanisms. The expression of alternative oxidase differed, increasing throughout development with protein peaking at day 20, perhaps suggesting a role in senescence. Overall, this study indicated that respiratory gene expression and protein abundance is co-ordinated with respiratory activity for most components but that some components, such as the F1beta subunit may be under discrete forms of regulation.


Prognostic factors associated with the response to sunitinib in patients with metastatic renal cell carcinoma.
We investigated the prognostic clinicopathologic factors associated with overall survival (os) and progression-free survival (pfs) in the once-daily continuous administration of first-line sunitinib in a consecutive cohort of Turkish patients with metastatic renal cell carcinoma (rcc). The study enrolled 77 Turkish patients with metastatic rcc who received sunitinib in a continuous once-daily dosing regimen between April 2006 and April 2011. Univariate analyses were performed using the log-rank test. Median follow-up was 18.5 months. In univariate analyses, poor pfs and os were associated with 4 of the 5 factors in the Memorial Sloan-Kettering Cancer Center (mskcc) score: Eastern Cooperative Oncology Group performance status of 2 or higher, low hemoglobin, high corrected serum calcium, and high lactate dehydrogenase. In addition to those factors, hypoalbuminemia, more than 2 metastatic sites, liver metastasis, non-clear cell histology, and the presence of sarcomatoid features on pathology were also associated with poor pfs; and male sex, hypoalbuminemia, prior radiotherapy, more than 2 metastatic sites, lung metastasis, nuclear grade of 3 or 4 for the primary tumour, and the presence of sarcomatoid features were also associated with poorer os. The application of the mskcc model distinctly separated the pfs and os curves (p < 0.001). Our study identified prognostic factors for pfs and os with the use sunitinib as first-line metastatic rcc therapy and confirmed that the mskcc model still appears to be valid for predicting survival in metastatic rcc in the era of molecular targeted therapy.


Prediction of Prophages and Their Host Ranges in Pathogenic and Commensal Neisseria Species.
The genus Neisseria includes two pathogenic species, N. gonorrhoeae and N. meningitidis, and numerous commensal species. Neisseria species frequently exchange DNA with one another, primarily via transformation and homologous recombination and via multiple types of mobile genetic elements (MGEs). Few Neisseria bacteriophages (phages) have been identified, and their impact on bacterial physiology is poorly understood. Furthermore, little is known about the range of species that Neisseria phages can infect. In this study, we used three virus prediction tools to scan 248 genomes of 21 different Neisseria species and identified 1,302 unique predicted prophages. Using comparative genomics, we found that many predictions are dissimilar from prophages and other MGEs previously described to infect Neisseria species. We also identified similar predicted prophages in genomes of different Neisseria species. Additionally, we examined CRISPR-Cas targeting of each Neisseria genome and predicted prophage. While CRISPR targeting of chromosomal DNA appears to be common among several Neisseria species, we found that 20% of the prophages we predicted are targeted significantly more than the rest of the bacterial genome in which they were identified (i.e., backbone). Furthermore, many predicted prophages are targeted by CRISPR spacers encoded by other species. We then used these results to infer additional host species of known Neisseria prophages and predictions that are highly targeted relative to the backbone. Together, our results suggest that we have identified novel Neisseria prophages, several of which may infect multiple Neisseria species. These findings have important implications for understanding horizontal gene transfer between members of this genus. IMPORTANCE Drug-resistant Neisseria gonorrhoeae is a major threat to human health. Commensal Neisseria species are thought to serve as reservoirs of antibiotic resistance and virulence genes for the pathogenic species N. gonorrhoeae and N. meningitidis. Therefore, it is important to understand both the diversity of mobile genetic elements (MGEs) that can mediate horizontal gene transfer within this genus and the breadth of species these MGEs can infect. In particular, few bacteriophages (phages) are known to infect Neisseria species. In this study, we identified a large number of candidate phages integrated in the genomes of commensal and pathogenic Neisseria species, many of which appear to be novel phages. Importantly, we discovered extensive interspecies targeting of predicted phages by Neisseria CRISPR-Cas systems, which may reflect their movement between different species. Uncovering the diversity and host range of phages is essential for understanding how they influence the evolution of their microbial hosts.


Single Residue Substitutions That Confer Voltage-Gated Sodium Ion Channel Subtype Selectivity in the NaV1.7 Inhibitory Peptide GpTx-1.
There is interest in the identification and optimization of new molecular entities selectively targeting ion channels of therapeutic relevance. Peptide toxins represent a rich source of pharmacology for ion channels, and we recently reported GpTx-1 analogs that inhibit NaV1.7, a voltage-gated sodium ion channel that is a compelling target for improved treatment of pain. Here we utilize multi-attribute positional scan (MAPS) analoging, combining high-throughput synthesis and electrophysiology, to interrogate the interaction of GpTx-1 with NaV1.7 and related NaV subtypes. After one round of MAPS analoging, we found novel substitutions at multiple residue positions not previously identified, specifically glutamic acid at positions 10 or 11 or lysine at position 18, that produce peptides with single digit nanomolar potency on NaV1.7 and 500-fold selectivity against off-target sodium channels. Docking studies with a NaV1.7 homology model and peptide NMR structure generated a model consistent with the key potency and selectivity modifications mapped in this work.


Masticatory stress, orbital orientation and the evolution of the primate postorbital bar.
A postorbital bar is one of a suite of derived features which distinguishes basal primates from their putative sister taxon, plesiadapiforms. Two hypotheses have been put forward to explain postorbital bar development and variation in circumorbital form: the facial torsion model and visual predation hypothesis. To test the facial torsion model, we employ strain data on circumorbital and mandibular loading patterns in representative primates with a postorbital bar and masticatory apparatus similar to basal primates. To examine the visual predation hypothesis, we employ metric data on orbit orientation in Paleocene and Eocene primates, as well as several clades of visual predators and foragers that vary interspecifically in postorbital bar formation.A comparison of galago circumorbital and mandibular peak strains during powerful mastication demonstrates that circumorbital strains are quite low. This indicates that, as in anthropoids, the strepsirhine circumorbital region is excessively overbuilt for countering routine masticatory loads. The fact that circumorbital peak-strain levels are uniformly low in both primate suborders undermines any model which posits that masticatory stresses are determinants of circumorbital form, function and evolution. This is interpreted to mean that sufficient cortical bone must exist to prevent structural failure due to non-masticatory traumatic forces. Preliminary data also indicate that the difference between circumorbital and mandibular strains is greater in larger taxa.Comparative analyses of several extant analogs suggest that the postorbital bar apparently provides rigidity to the lateral orbital margins to ensure a high level of visual acuity during chewing and biting. The origin of the primate postorbital bar is linked to changes in orbital convergence and frontation at smaller sizes due to nocturnal visual predation and increased encephalization. By incorporating in vivo and fossil data, we reformulate the visual predation hypothesis of primate origins and thus offer new insights into major adaptive transformations in the primate skull.


Occurrence of Shiga toxin-producing Escherichia coli O157 in selected dairy and meat products marketed in the city of Rabat, Morocco.
Samples of meat and dairy products taken from the city of Rabat, Morocco, were examined for the presence of Escherichia coli O157 by the selective enrichment procedure followed by plating on cefixime-tellurite-sorbitol MacConkey agar and a latex agglutination test. The ability of isolates to produce Shiga toxins (ST1 or ST2) was also tested by an agglutination test using sensitized latex. Dairy samples (n = 44) included different products commonly consumed in the country. Meat samples (n = 36) were taken from traditional butchers because these products are generally marketed in this way. Random samples were taken from each product during the period of January through May. Of the 80 samples tested, 8 (10%) harbored E. coli O157. Four dairy and four meat samples were contaminated (9.1 and 11.1%, respectively). Of 10 E. coli O157 isolates from contaminated samples demonstrating true antigen-antibody agglutination, 5 (50%) produced either ST2 alone or ST2 plus ST1. Four of the five strains (80%) were meat isolates and produced ST2 with or without ST1, and the fifth was a dairy isolate producing ST2.


Mapping the DNA Damaging Effects of Polypyridyl Copper Complexes with DNA Electrochemical Biosensors.
Several classes of copper complexes are known to induce oxidative DNA damage that mediates cell death. These compounds are potentially useful anticancer agents and detailed investigation can reveal the mode of DNA interaction, binding strength, and type of oxidative lesion formed. We recently reported the development of a DNA electrochemical biosensor employed to quantify the DNA cleavage activity of the well-studied [Cu(phen)2]2+ chemical nuclease. However, to validate the broader compatibility of this sensor for use with more diverse-and biologically compatible-copper complexes, and to probe its use from a drug discovery perspective, analysis involving new compound libraries is required. Here, we report on the DNA binding and quantitative cleavage activity of the [Cu(TPMA)(N,N)]2+ class (where TPMA = tris-2-pyridylmethylamine) using a DNA electrochemical biosensor. TPMA is a tripodal copper caging ligand, while N,N represents a bidentate planar phenanthrene ligand capable of enhancing DNA interactions through intercalation. All complexes exhibited electroactivity and interact with DNA through partial (or semi-) intercalation but predominantly through electrostatic attraction. Although TPMA provides excellent solution stability, the bulky ligand enforces a non-planar geometry on the complex, which sterically impedes full interaction. [Cu(TPMA)(phen)]2+ and [Cu(TPMA)(DPQ)]2+ cleaved 39% and 48% of the DNA strands from the biosensor surface, respectively, while complexes [Cu(TPMA)(bipy)]2+ and [Cu(TPMA)(PD)]2+ exhibit comparatively moderate nuclease efficacy (ca. 26%). Comparing the nuclease activities of [Cu(TPMA)(phen)] 2+ and [Cu(phen)2]2+ (ca. 23%) confirms the presence of TPMA significantly enhances chemical nuclease activity. Therefore, the use of this DNA electrochemical biosensor is compatible with copper(II) polypyridyl complexes and reveals TPMA complexes as a promising class of DNA damaging agent with tuneable activity due to coordinated ancillary phenanthrene ligands.


Nuclear factor (NF)-κB and its associated pathways are major molecular regulators of blood-induced joint damage in a murine model of hemophilia.
The present study was designed to investigate the molecular signaling events from onset of bleeding through the development of arthropathy in a murine model of hemophilia A. A sharp-injury model of hemarthrosis was used. A global gene expression array on joint-specific RNA isolated 3 h post-injury revealed nuclear factor-kappa B (NF-κB) as the major transcription factor triggering inflammation. As a number of genes encoding the cytokines, growth factors and hypoxia regulating factors are known to be activated by NF-κB and many of these are part of the pathogenesis of various joint diseases, we reasoned that NF-κB-associated pathways may play a crucial role in blood-induced joint damage. To further understand its role, we screened NF-κB-associated pathways between 1 h to 90 days after injury. After a single articular bleed, distinct members of the NF-κB family (NF-κB1/NF-κB2/RelA/RelB) and their responsive pro-inflammatory cytokines (IL-1β/IL-6/IFNγ/TNFα) were significantly up-regulated (> 2 fold, P < 0.05) in injured vs. control joints at the various time-points analyzed (1 h/3 h/7 h/24 h). After multiple bleeds (days 30/60/75/90), there was increased expression of NF-κB-associated factors that contribute to hypoxia (HIF-1α, 3.3-6.5 fold), angiogenesis (VEGF-α, 2.5-4.4 fold) and chondrocyte damage (matrix metalloproteinase-13, 2.8-3.8 fold) in the injured joints. Micro RNAs (miR) that are known to regulate NF-κB activation (miRs-9 and 155), inflammation (miRs-16, 155 and 182) and apoptosis (miRs-19a, 155 and 186) were also differentially expressed (-4 to +13-fold) after joint bleeding, indicating that the small RNAs could modulate the arthropathy phenotype. These data suggest that NF-κB-associated signaling pathways are involved in the development of hemophilic arthropathy.


Burying beetles regulate the microbiome of carcasses and use it to transmit a core microbiota to their offspring.
Necrophagous beetles utilize carrion, a highly nutritious resource that is susceptible to intense microbial competition, by treating it with antimicrobial anal and oral secretions. However, how this regulates the carcass microbiota remains unclear. Here, we show that carcasses prepared by the burying beetle Nicrophorus vespilloides undergo significant changes in their microbial communities subsequent to their burial and "preparation." Prepared carcasses hosted a microbial community that was more similar to that of beetles' anal and oral secretions than to the native carcass community or the surrounding soil, indicating that the beetles regulated the carcass microbiota. A core microbial community (Xanthomonadaceae, Enterococcaceae, Enterobacteriaceae and Yarrowia yeasts) was transmitted by the beetles to the larvae via the anal and oral secretions and the carcass surface. These core taxa proliferated on the carcass, indicating a growth conducive environment for these microbes when associated with beetles. However, total bacterial loads were higher on decomposing carcasses without beetles than on beetle-prepared carcasses, indicating that the beetles and/or their associated symbionts suppress the growth of competing microbes. Thus, apart from being a nutritional resource, the carcass provides a medium for vertical transmission of a tightly regulated symbiotic microbiota, whose activity on the carcass and in the larval gut may involve carcass preservation as well as digestion.


Nurses' knowledge of pharmacological measures on acute pain management in Western Saudi Arabia.
To assess the nursing knowledge of acute pain management in a tertiary hospital. Three hundred closed-answer questionnaires were distributed in various hospital departments at King Khalid National Guard Hospital in Jeddah, Saudi Arabia in 2007. Three main topics were tested in the questionnaire; nursing opinion on patient self-report of pain as a main indicator of pain intensity, the need to increase opioids dose when the first dose had been safe but ineffective, and nurses' attitude toward the incidence of addiction, tolerance, and physical dependence. Of the respondents, 45.8% used the patient self-report of pain as an indicator of pain intensity in the smiling patient (A), and 55.1% relied on that in the grimacing patient (B). Fifty percent of respondents in patient A and 30.5% in patient B decided to give no more morphine to both patients despite their pain, while 7.6% and 19.5% of nurses selected the option of giving higher morphine dose to both patients. Only 38.1% of nurses chose the correct answer for risk of addiction (<1%) and 41.6% selected an exaggerated response range from 25-100% as a chance of addiction. Very few nurses recognized the problem of tolerance and physical dependence and picked the right answers. Nurses' knowledge of acute pain management is deficient in many aspects. Proper education is needed to improve their knowledge of pain.


Metabolism in patients with small cell lung carcinoma compared with patients with non-small cell lung carcinoma and healthy controls.
Weight loss is a frequently occurring problem in patients with lung cancer due to an increased resting energy expenditure (REE) and a decreased energy intake. The aim of the present study was to compare the metabolic and inflammatory characteristics of patients with small cell lung carcinoma (SCLC) and non-small cell lung carcinoma (NSCLC). The metabolic parameters of the lung cancer population were compared with those of a healthy control group. REE was measured in 66 patients with lung cancer, subdivided according to their histology, and in 33 healthy controls matched for sex, age, and fat free mass (FFM). Inflammatory mediators were measured in the plasma of the patients with lung cancer. An increased REE adjusted for FFM was found in the patients with lung cancer. Those with small cell lung carcinoma (SCLC) had an increased REE adjusted for FFM (mean 1925 kcal/day) compared with those with non-small cell lung carcinoma (NSCLC) (mean 1789 kcal/day, 95% CI for difference 36 to 236). FFM accounted for 69% and 48% of the inter-individual variation in REE in controls and those with NSCLC, respectively, while FFM accounted for only 25% of the variation in REE in patients with SCLC in whom the fat mass (FM) also contributed significantly (28%) to the variation in REE. Increased concentrations of soluble TNF-receptor 75 (sTNF-R75) and cortisol were found in patients with SCLC compared with those with NSCLC. Lipopolyasccharide binding protein (LBP) and sTNF-R55 were related to plasma levels of cortisol. An enhanced REE adjusted for FFM occurred in patients with SCLC compared with those with NSCLC.


Teasing apart the host-related, nutrient-related and temperature-related effects shaping the phenology and microbiome of the tropical seagrass Halophila stipulacea.
Halophila stipulacea seagrass meadows are an ecologically important and threatened component of the ecosystem in the Gulf of Aqaba. Recent studies have demonstrated correlated geographic patterns for leaf epiphytic community composition and leaf morphology, also coinciding with different levels of water turbidity and nutrient concentrations. Based on these observations, workers have suggested an environmental microbial fingerprint, which may reflect various environmental stress factors seagrasses have experienced, and may add a holobiont level of plasticity to seagrasses, assisting their acclimation to changing environments and through range expansion. However, it is difficult to tease apart environmental effects from host-diversity dependent effects, which have covaried in field studies, although this is required in order to establish that differences in microbial community compositions among sites are driven by environmental conditions rather than by features governed by the host. In this study we carried out a mesocosm experiment, in which we studied the effects of warming and nutrient stress on the composition of epiphytic bacterial communities and on some phenological traits. We studied H. stipulacea collected from two different meadows in the Gulf of Aqaba, representing differences in the host and the environment alike. We found that the source site from which seagrasses were collected was the major factor governing seagrass phenology, although heat increased shoot mortality and nutrient loading delayed new shoot emergence. Bacterial diversity, however, mostly depended on the environmental conditions. The most prominent pattern was the increase in Rhodobacteraceae under nutrient stress without heat stress, along with an increase in Microtrichaceae. Together, the two taxa have the potential to maintain nitrate reduction followed by an anammox process, which can together buffer the increase in nutrient concentrations across the leaf surface. Our results thus corroborate the existence of environmental microbial fingerprints, which are independent from the host diversity, and support the notion of a holobiont level plasticity, both important to understand and monitor H. stipulacea ecology under the changing climate.


Effects of adding fat to the sow lactation diet on lactation and rebreeding performance.
One-hundred-three multiparous sows were randomly assigned to one of two lactation diets containing either no supplemental animal fat (C) or 10% added fat (F) during two seasons, summer (S) and winter (W), in a 2 X 2 factorial arrangement of treatments. Sows were placed on their respective dietary treatments 1 wk prior to farrowing and were fed these diets ad libitum throughout the 28-d lactation period. Weekly feed intake and total feed intake were not affected by diet or season, while weekly metabolizable energy (ME) intake tended to be higher during week 1 and 3, and was higher (P less than .04) during wk 2 for sows fed diet F. Sow weight loss from farrowing to 21 d of lactation and to weaning (28 d) were unaffected by diet or season. Average pig birth weight was .15 kg higher (P less than .01) for pigs born during S compared with those born in W. Sows receiving diet F had heavier litters at 21 d (P less than .01) and heavier average pig 21-d weights (P less than .01). This was primarily due to the 13.1% increase (P less than .04) in estimated milk yield and the higher fat concentration (P less than .001) of milk consumed by the pigs nursing sows fed diet F. Interval between weaning and rebreeding was shortened by 5.9 d (P less than .01) for sows during W than during S, and tended to be lower for sows fed diet F (7.3 d) compared with that of sows fed diet C (9.7 d).(ABSTRACT TRUNCATED AT 250 WORDS)


Development of surface polarity in mouse eggs.
Investigations were carried out to determine what effects components of the cytoskeletal system and meiotic spindle have on the development and maintenance of surface polarity in mouse ova. The surface of the mature egg possessed numerous microvilli except for a region (microvillus-free area) adjacent to the meiotic spindle. In contrast, the surface of the immature oocyte was covered uniformly with a dense population of microvilli. When cultured in vitro immature oocytes spontaneously underwent maturation; a meiotic spindle formed in the center of the ovum which then moved to the cortex. Coincident with the cortical localization of the meiotic spindle was the formation of a microvillus-free area and subjacent layer of microfilaments. A microvillus-free area did not form when meiotic maturation was inhibited with dibutyryl cyclic AMP or chloroquine. If immature oocytes were incubated in cytochalasin B a meiotic spindle developed, but it did not become localized to the egg cortex and a microvillus-free area failed to form. Oocytes incubated in colchicine underwent germinal vesicle breakdown and chromosome condensation, a meiotic spindle did not form but the chromosomes became localized to the ovum cortex where a microvillus-free area developed. These results and observations of mature ova treated with cytochalasin B or colchicine indicate that mechanisms involving the movement of the meiotic spindle to the oocyte cortex and development and maintenance of surface polarity are cytochalasin B sensitive. Cortical localization of meiotic chromosomes brings about the formation of a microvillus-free area.


Comparative purification strategies for Drosophila and human mitochondrial DNA replication proteins: DNA polymerase gamma and mitochondrial single-stranded DNA-binding protein.
The mitochondrion is the eukaryotic organelle that carries out oxidative phosphorylation, fulfilling cellular requirements for ATP production. Disruption of mitochondrial energy metabolism can occur by genetic and biochemical mechanisms involving nuclear-encoded proteins that are required at the mitochondrial DNA replication fork, which often leads to human disorders and to animal lethality during development. DNA polymerase gamma (pol gamma), the mitochondrial replicase, and the mitochondrial single-stranded DNA-binding protein (mtSSB) have been the focus of study in our lab for a number of years. Here we describe the purification strategies that we developed for obtaining the recombinant forms of pol gamma and mtSSB from both Drosophila melanogaster and humans. Despite the fact that similar approaches can be used for purifying the homologous proteins, we have observed that there are differences in the behavior of the proteins in some specific steps that may reflect differences in their structural and biochemical properties. Their purification in homogeneous, active form represents the first step toward our long-term goal to understand their biochemistry, biology, and functions at the mitochondrial DNA replication fork.


Human brain glial cells synthesize thrombospondin.
Thrombospondin, a 450-kDa multinodular glycoprotein with lectin-type activity, is found in human platelets, endothelial cells, fibroblasts, smooth muscle cells, monocytes, and granular pneumocytes. Thrombospondin interacts with heparin, fibrinogen, fibronectin, collagen, histidine-rich glycoprotein, and plasminogen. Recently, thrombospondin synthesis by smooth muscle cells has been reported to be augmented by platelet-derived growth factor. We present evidence that thrombospondin is present within and synthesized by astrocytic neuroglial cells. Heparin-Sepharose affinity chromatography of material derived from a human brain homogenate yielded a protein that, when reduced, had an apparent size of 180 kDa and comigrated with reduced platelet thrombospondin on NaDodSO4/PAGE. Immunoblot analysis with monospecific anti-thrombospondin confirmed the presence of immunoreactive thrombospondin. Indirect immunofluorescence of cultured human glial cells indicated the presence of thrombospondin. Metabolic labeling of glial cell cultures with [35S]methionine followed by immunoprecipitation with monospecific anti-thrombospondin revealed synthesis of a 180-kDa polypeptide that comigrated with platelet thrombospondin on NaDodSO4/PAGE. Cultured human glial cells were incubated for 48 hr in serum-free medium with purified platelet-derived growth factor at concentrations up to 50 ng/ml. Aliquots taken at intervals were analyzed by a quantitative double-antibody ELISA. The growth factor stimulated the release of thrombospondin into the culture medium by as much as 10-fold over control cultures. The presence of thrombospondin within glial cells of the central nervous system and the augmentation of its synthesis by platelet-derived growth factor suggest that thrombospondin may play an important role in regulating cell-cell and cell-matrix interactions during periods of cell division and growth.


[Subdural, extra-arachnoid block as a complication of stellate ganglion block: documentation with ultrasound].
We present a patient who developed a high central neural block after stellate-ganglion-blockade. The underlying pathophysiology was assessed via sonographic imaging. Stellate ganglion block was performed in a 19-year old patient according to the standard technique. Multiple aspiration tests were negative and a test dose of 3 ml bupivacaine 0.25% was injected. After a 3 min interval another 5 ml were injected. Two minutes after the local anaesthetic administration the patient reported nausea and sensations in the upper extremity. Spontaneous respiration efforts stopped, and the patient became unconscious. Tracheal intubation was performed, and the patient was ventilated in a controlled mode for two hours. Heart rate as well as blood pressure remained within the normal range. Neurologic recovery occurred rapidly and extubation was performed about two and a half hours after the event. Our sonographic studies demonstrated a local anaesthetic depot directly at the root of C 6, with a mean diameter of 10 mm and a length of 5 to 6 cm (about a third smaller than expected). Sonographic studies and clinical symptoms of our patient are most likely to occur with a subdural extra-arachnoidal block. Ultrasonographic guided puncture enhances the patient's safety by the opportunity to directly visualise transverse process, nerval root as well as local anaesthetic depot. In case of depot formation directly at the nerval root, injection may be stopped and the needle repositioned. Furthermore, direct visualisation of the great vessels (A. vertebralis) prevents intravascular injection and haematoma formation.


[Facial paralysis of obstetrical origin. Etiology, evolution].
In the case of 17 obstetrical facial paralyses occurring in a total of 20,409 births, the authors have studied the circumstances of their appearance and the prognosis for this lesion. It is a rare lesion favoured by asymmetric purchase when using articulated forceps. The authors insist that use of Thierry's forceps do not harm the facial nerve. They emphasise the possibility that progression may not always be favourable.


IS10 promotes adjacent deletions at low frequency.
Some transposable elements move by a replicative mechanism involving cointegrate formation. Intramolecular cointegration can generate a product called an "adjacent deletion" in which a contiguous chromosomal segment adjacent to the transposon is deleted while the element responsible remains intact. Insertion sequence IS10 is thought to transpose by a nonreplicative mechanism. In the simplest models, nonreplicative transposition cannot give rise to an adjacent deletion because an intrinsic feature of such transposition is excision of the IS element from the donor location. We report here that IS10 can generate adjacent deletions, but at a frequency which is approximately 1/30th the frequency of transposition for the same element. We suggest that these deletions might arise either by nonreplicative transposition events that involve two IS10 elements located on sister chromosomes or by aberrant nonreplicative events involving cleavage and ligation at only one end of the element.


Environmental factors responsible for switching on the SO₄²⁻ excretory system in the kidney of seawater eels.
Eels are unique in that they maintain lower plasma SO(4)(2-) concentration in SO(4)(2-)-rich (∼30 mM) seawater (SW) than in SO(4)(2-)-poor (<0.3 mM) freshwater (FW), showing drastic changes in SO(4)(2-) regulation between FW and SW. We previously showed that the expression of renal SO(4)(2-) transporter genes, FW-specific Slc13a1 and SW-specific Slc26a6a, changes profoundly after transfer of FW eels to SW, which results in the decrease in plasma SO(4)(2-) concentration after 3 days in SW. In this study, we attempted to identify the environmental factor(s) that trigger the switching of SO(4)(2-) regulation using changes in plasma and urine SO(4)(2-) concentrations and expression of the transporter genes as markers. Transfer of FW eels to 30 mM SO(4)(2-) or transfer of SW eels to SO(4)(2-)-free SW did not change the SO(4)(2-) regulation. Major divalent cations in SW, Mg(2+) (50 mM) and Ca(2+) (10 mM), were also ineffective, but 50 mM NaCl was effective for switching the SO(4)(2-) regulation. Further analyses using choline-Cl and Na-gluconate showed that Cl(-) is a primary factor and Na(+) is permissive for the Cl(-) effect. Since plasma SO(4)(2-) and Cl(-) concentrations were inversely correlated, we injected various solutions into the blood and found that Cl(-) alone triggered the switching from FW to SW-type regulation. Furthermore, the inhibitor of Na-Cl cotransporter (NCC) added to media significantly impaired the expression of SW-specific Slc26a6a in 150 mM NaCl. In summary, it appears that Cl(-) ions in SW are taken up into the circulation via the NCC together with Na(+), and the resultant increase in plasma Cl(-) concentration enhances SO(4)(2-) excretion by the kidney through downregulation of absorptive Slc13a1 and upregulation of excretory Slc26a6a, resulting in low plasma SO(4)(2-) concentration in SW.


PCBP2 knockdown promotes ferroptosis in malignant mesothelioma.
Malignant mesothelioma (MM) is still increasing worldwide. The pathogenesis depends on asbestos-induced iron accumulation, which eventually leads to ferroptosis-resistance of mesothelial cells via somatic mutations. Poly (rC)-binding proteins 1 and 2 (PCBP1/2) are recently recognized cytosolic Fe(II) chaperones. Here we studied the role of PCBP1/2 in rat/human mesothelial and MM cells as well as rat/human MM specimens. Normal peritoneal mesothelial cells in rats exhibited PCBP1 but not PCBP2 immunopositivity whereas primary/immortalized mesothelial cells showed PCBP1/2 immunopositivity. Rat MM specimens induced by intraperitoneal injection of chrysotile, including in situ lesion, revealed PCBP1/2 immunopositivity (90% for both) in the nucleus and cytoplasm with a tendency of higher expression in epithelioid subtype. Knockdown of PCBP2 but not PCBP1 significantly decreased both TfR1 and FTH expression in MM cells with inhibition of proliferation, indicating stagnation of intracellular iron transport. Erastin, a cysteine-deprivation type ferroptosis inducer, decreased the expression of both PCBP1/2 in MM cells. Furthermore, PCBP2 knockdown significantly increased the sensitivity of MM cells to erastin-induced ferroptosis with increased catalytic Fe(II). In conclusion, PCBP2 works for ferroptosis-resistance not only during mesothelial carcinogenesis but also in MM, which warrants further investigation as a novel therapeutic target.


Stochastic interference in a dispersive nonlinear optical fiber system.
Stochastic power fluctuation in a fiber optic system due to the interplay among dispersion, nonlinearity and partial coherence of the source is investigated. An analytical expression for the power fluctuation of a signal pulse due to its interference with an echo pulse generated due to the nonlinear interaction of signal pulses in a fiber optic system excited by a partially coherent source is obtained. The analytical results show that the mean nonlinear distortion decreases as the coherence time of the source reduces is consistent with our numerical simulations.


Mapping the interaction sites of Mucin 1 and DNA aptamer by atomic force microscopy.
Mucin 1 (MUC1) is an attractive tumor marker for cancer diagnosis. An advanced atomic force microscopy (AFM) mode, peak-force tapping AFM with an aptamer functionalized tip, was introduced to map the specific interaction sites of an aptamer and MUC1. Single molecular force spectroscopy (SMFS) was used to investigate dynamic parameters of the aptamer-MUC1.


Current status of Anopheles stephensi response to various insecticides in some areas of the Thar desert.
Investigations on the current response of A. stephensi. to six insecticides viz. DDT, dieldrin, malathion, fenitrothion, propoxur and permethrin, were carried out in 3 districts i.e. Barmer, Jodhpur and Pali, of the Thar desert. The species was found resistant to DDT and dieldrin, partially resistant to malathion and susceptible to fenitrothion, propoxur and permethrin. Dieldrin and malathion resistance has been detected for the first time in the Thar desert. Lethal concentrations (LC50 & LC95) of DDT and dieldrin and lethal exposure times (LT50 & LT95) of malathion, fenitrothion, propoxur and permethrin have been determined. In some areas, the differences in LC50 and LT50 values of tested insecticides, except fenitrothion, were found statistically significant. Chi-square and regression tests have revealed the homogeniety and linear trend respectively in the response of A. stephensi to insecticides. The findings of the study indicate that organochlorine compounds can be used alternately in the spray operations.


Ras-driven transcriptome analysis identifies aurora kinase A as a potential malignant peripheral nerve sheath tumor therapeutic target.
Patients with neurofibromatosis type 1 (NF1) develop malignant peripheral nerve sheath tumors (MPNST), which are often inoperable and do not respond well to current chemotherapies or radiation. The goal of this study was to use comprehensive gene expression analysis to identify novel therapeutic targets. Nerve Schwann cells and/or their precursors are the tumorigenic cell types in MPNST because of the loss of the NF1 gene, which encodes the RasGAP protein neurofibromin. Therefore, we created a transgenic mouse model, CNP-HRas12V, expressing constitutively active HRas in Schwann cells and defined a Ras-induced gene expression signature to drive a Bayesian factor regression model analysis of differentially expressed genes in mouse and human neurofibromas and MPNSTs. We tested functional significance of Aurora kinase overexpression in MPNST in vitro and in vivo using Aurora kinase short hairpin RNAs (shRNA) and compounds that inhibit Aurora kinase. We identified 2,000 genes with probability of linkage to nerve Ras signaling of which 339 were significantly differentially expressed in mouse and human NF1-related tumor samples relative to normal nerves, including Aurora kinase A (AURKA). AURKA was dramatically overexpressed and genomically amplified in MPNSTs but not neurofibromas. Aurora kinase shRNAs and Aurora kinase inhibitors blocked MPNST cell growth in vitro. Furthermore, an AURKA selective inhibitor, MLN8237, stabilized tumor volume and significantly increased survival of mice with MPNST xenografts. Integrative cross-species transcriptome analyses combined with preclinical testing has provided an effective method for identifying candidates for molecular-targeted therapeutics. Blocking Aurora kinases may be a viable treatment platform for MPNST.


The Prevalence of Delirium in An Older Acute Surgical Population and Its Effect on Outcome.
With an ageing population, an increasing number of older adults are admitted for assessment to acute surgical units. Older adults have specific factors that may influence outcomes, one of which is delirium (acute cognitive impairment). To establish the prevalence of delirium on admission in an older acute surgical population and its effect on mortality. Secondary outcomes investigated include hospital readmission and length of hospital stay. This observational multi-centre study investigated consecutive patients, ≥65 years, admitted to the acute surgical units of five UK hospitals during an eight-week period. On admission the Confusion Assessment Method (CAM) score was performed to detect delirium. The effect of delirium on important clinical outcomes was investigated using tests of association and logistic regression models. The cohort consisted of 411 patients with a mean age of 77.3 years (SD 8.1). The prevalence of admission delirium was 8.8% (95% CI 6.2-11.9%) and cognitive impairment was 70.3% (95% CI 65.6-74.7%). The delirious group were not more likely to die at 30 or 90 days (OR 1.1, 95% CI 0.2 to 5.1, p = 0.67; OR 1.4, 95% CI 0.4 to 4.1. p = 0.82) or to be readmitted within 30 days of discharge (OR 0.9, 95% CI 0.4 to 2.2, p = 0.89). Length of hospital stay was significantly longer in the delirious group (median 8 vs. 5 days respectively, p = 0.009). Admission delirium occurs in just under 10% of older people admitted to acute surgical units, resulting in significantly longer hospital stays.


A morphologic and fluorophotometric analysis of the corneal endothelium in type I diabetes mellitus and cystic fibrosis.
Seven patients with type I diabetes mellitus (Group 1), seven with normoglycemic cystic fibrosis (Group 2), seven with hyperglycemic cystic fibrosis (Group 3), and ten age-matched control subjects underwent corneal fluorophotometry and quantitative specular microscopy. Group 1 had background microangiopathic retinopathy but no evidence of proliferative disease by fluorescein angiography. Significant increases in mean corneal endothelial permeability and mean pump rate occurred in Group 1, indicating a defect in the endothelial barrier function early in type I diabetes mellitus. Similar significant increases in mean corneal endothelial permeability and mean pump rate occurred in both cystic fibrosis groups. The greatest increase was found in Group 3, suggesting a primary defect in the endothelial barrier function in cystic fibrosis, aggravated by the hyperglycemic state. No morphologic abnormalities were noted in Group 1, but both cystic fibrosis groups had smaller mean cell areas than did the control group. There were significant differences in the morphologic and functional correlations between Groups 1 and 3, suggesting different mechanisms for the increased endothelial permeability in these two disorders.


Production of metallothionein and heat-shock proteins in response to metals.
Acute stress, such as heat, and some metals, such as arsenite, will induce a specific group of stress proteins referred to as heat-shock proteins. The heat-shock proteins contribute to the survival of cells following a variety of stresses. Similarly, metals such as cadmium and zinc, will increase the levels of metallothionein (MT). The metal-binding protein, MT, has also been found to have a protective role in the cellular response to acute stresses like heavy metals. The purpose of the present study was to examine the production of these proteins in response to metals. Rat hepatocytes were maintained in monolayer culture for 22 hr, and subsequently treated with various concentrations of metals for 4 hr, or incubated at 43.5 degrees C for 15-60 min. Following two washes with fresh media, the cells were labeled with [35S]-methionine (25 microCi/ml) in methionine-free media for 4 hr for determination of heat-shock protein production, or reincubated in fresh media for 20 hr for MT determination. Heat-shock protein production was determined by SDS-polyacrylamide gel electrophoresis followed by autoradiography. The autoradiograms were quantified by densitometric scanning. MT was determined by the Cd/hemoglobin affinity assay. Three metals (arsenite, cadmium, and zinc) strongly increased the heat-shock proteins. Whereas arsenite was a much less effective inducer of MT than was cadmium or zinc, arsenite was as effective as the other metals in inducing heat-shock proteins. Nickel was a good inducer of MT; however, it resulted in only a slight increase in the levels of heat-shock proteins.(ABSTRACT TRUNCATED AT 250 WORDS)


Junctional Adhesion Molecules in Cancer: A Paradigm for the Diverse Functions of Cell-Cell Interactions in Tumor Progression.
Tight junction (TJ) proteins are essential for mediating interactions between adjacent cells and coordinating cellular and organ responses. Initial investigations into TJ proteins and junctional adhesion molecules (JAM) in cancer suggested a tumor-suppressive role where decreased expression led to increased metastasis. However, recent studies of the JAM family members JAM-A and JAM-C have expanded the roles of these proteins to include protumorigenic functions, including inhibition of apoptosis and promotion of proliferation, cancer stem cell biology, and epithelial-to-mesenchymal transition. JAM function by interacting with other proteins through three distinct molecular mechanisms: direct cell-cell interaction on adjacent cells, stabilization of adjacent cell surface receptors on the same cell, and interactions between JAM and cell surface receptors expressed on adjacent cells. Collectively, these diverse interactions contribute to both the pro- and antitumorigenic functions of JAM. In this review, we discuss these context-dependent functions of JAM in a variety of cancers and highlight key areas that remain poorly understood, including their potentially diverse intracellular signaling networks, their roles in the tumor microenvironment, and the consequences of posttranslational modifications on their function. These studies have implications in furthering our understanding of JAM in cancer and provide a paradigm for exploring additional roles of TJ proteins.


Colorectal Cancer Screening in a Nationwide High-deductible Health Plan Before and After the Affordable Care Act.
Little is known about the effect of the Affordable Care Act's (ACA) elimination of out-of-pocket costs for preventive services. This policy likely reduced out-of-pocket colonoscopy costs most for high-deductible health plan (HDHP) members. Determine the ACA's impact on colorectal cancer screening among HDHP members. Pre-post with comparison group, constructed before and after the ACA. We studied 2003-2012 administrative claims data of a large national health insurer. HDHP members had 1 year of low-deductible (≤$500) plan enrollment followed by 1 year of HDHP (≥$1000) enrollment after an employer-mandated switch; HDHP enrollment occurred fully after the ACA for 21,605 members and fully before the ACA for 106,609 members. We propensity score-matched contemporaneous low-deductible (≤$500) control group members to both the before-ACA and after-ACA HDHP groups. We examined the 1-year impact of the HDHP switch separately in the before-ACA and after-ACA study cohorts, then compared these changes to estimate ACA effects. Overall colorectal cancer screening, colonoscopy, and fecal-occult blood testing annual rates. Before the ACA, colorectal cancer screening tests declined by 37/10,000 (-71, -4) among HDHP members versus controls; after the ACA, HDHP members experienced a nonsignificant increase in screening [+52/10,000 (-19,124)]. Corresponding changes in colonoscopy were -55/10,000 (-81, -29) before and +20/10,000 (-38, 78) after the ACA. Thus, the ACA was associated with increased colorectal cancer screening rates [+89/10,000 (11, 168); relative: +9.1% (0.5-17.8)] and screening colonoscopies [+75/10,000 (12-139); relative: +16.4% (2.5-30.3)] among HDHP members. The ACA was associated with improved colorectal cancer screening among HDHP members.


Strategies to inhibit entry of HBV and HDV into hepatocytes.
Although there has been much research into the pathogenesis and treatment of hepatitis B virus (HBV) and hepatitis D virus (HDV) infections, we still do not completely understand how these pathogens enter hepatocytes. This is because in vitro infection studies have only been performed in primary human hepatocytes. Development of a polarizable, HBV-susceptible human hepatoma cell line and studies of primary hepatocytes from Tupaia belangeri have provided important insights into the viral and cellular factors involved in virus binding and infection. The large envelope (L) protein on the surface of HBV and HDV particles has many different functions and is required for virus entry. The L protein mediates attachment of virions to heparan sulfate proteoglycans on the surface of hepatocytes. The myristoylated N-terminal preS1 domain of the L protein subsequently binds to the sodium taurocholate cotransporting polypeptide (NTCP, encoded by SLC10A1), the recently identified bona fide receptor for HBV and HDV. The receptor functions of NTCP and virus entry are blocked, in vitro and in vivo, by Myrcludex B, a synthetic N-acylated preS1 lipopeptide. Currently, the only agents available to treat chronic HBV infection target the viral polymerase, and no selective therapies are available for HDV infection. It is therefore important to study the therapeutic potential of virus entry inhibitors, especially when combined with strategies to induce immune-mediated killing of infected hepatocytes.


Determination of human serum alpha1-acid glycoprotein and albumin binding of various marketed and preclinical kinase inhibitors.
There are about 380 protein kinase inhibitors in drug development as of today and 15 drugs have been marketed already for the treatment of cancer. This time 139 validated kinase targets are in the focus of drug research of pharmaceutical companies and big efforts are made for the development of new, druglike kinase inhibitors. Plasma protein binding is an important factor of the ADME profiling of a drug compound. Human serum albumin (HSA) and alpha(1)-acid glycoprotein (AAG) are the most relevant drug carriers in blood plasma. Since previous literature data indicated that AAG is the principal plasma binding component of some kinase inhibitors the present work focuses on the comprehensive evaluation of AAG binding of a series of marketed and experimental kinase inhibitors by using circular dichroism (CD) spectroscopy approach. HSA binding was also evaluated by affinity chromatography. Protein binding interactions of twenty-six kinase inhibitors are characterized. The contribution of AAG and HSA binding data to the pharmacokinetic profiles of the investigated therapeutic agents is discussed. Structural, biological and drug binding properties of AAG as well as the applicability of the CD method in studying drug-protein binding interactions are also briefly reviewed.


Steroid receptor coactivator-1 from brain physically interacts differentially with steroid receptor subtypes.
In vitro studies reveal that nuclear receptor coactivators enhance the transcriptional activity of steroid receptors, including estrogen (ER) and progestin receptors (PR), through ligand-dependent interactions. Whereas work from our laboratory and others shows that steroid receptor coactivator-1 (SRC-1) is essential for efficient ER and PR action in brain, very little is known about receptor-coactivator interactions in brain. In the present studies, pull-down assays were used to test the hypotheses that SRC-1 from hypothalamic and hippocampal tissue physically associate with recombinant PR or ER in a ligand-dependent manner. SRC-1, from hypothalamus or hippocampus, interacted with PR-A and PR-B in the presence of an agonist, but not in the absence of ligand or in the presence of a selective PR modulator, RU486. Interestingly, SRC-1 from brain associated more with PR-B, the stronger transcriptional activator, than with PR-A. In addition, SRC-1 from brain, which was confirmed by mass spectrometry, interacted with ERalpha and ERbeta in the presence of agonist but not when unliganded or in the presence of the selective ER modulator, tamoxifen. Furthermore, SRC-1 from hypothalamus, but not hippocampus, interacted more with ERalpha than ERbeta, suggesting distinct expression patterns of other cofactors in these brain regions. These findings suggest that interactions of SRC-1 from brain with PR and ER are dependent on ligand, receptor subtype, and brain region to manifest the pleiotropic functional consequences that underlie steroid-regulated behaviors. The present findings reveal distinct contrasts with previous cell culture studies and emphasize the importance of studying receptor-coactivator interactions using biologically relevant tissue.


ECG dispersion mapping predicts clinical deterioration, measured by increase in the Simple Clinical Score.
ECG dispersion mapping (ECG-DM) is a novel technique that reports abnormal ECG microalternations. We report the ability of ECG-DM to predict clinical deterioration of acutely ill medical patients, as measured by an increase in the Simple Clinical Score (SCS) the day after admission to hospital. 453 acutely ill medical patients (mean age 69.7 +/- 14.0 years) had the SCS recorded and ECGDM performed immediately after admission to hospital. 46 patients had an SCS increase 20.8 +/- 7.6 hours after admission. Abnormal micro-alternations during left ventricular re-polarization had the highest association with SCS increase (p=0.0005). Logistic regression showed that only nursing home residence and abnormal micro-alternations during re-polarization of the left ventricle were independent predictors of SCS increase with an odds ratio of 2.84 and 3.01, respectively. ECG-DM changes during left ventricular re-polarization are independent predictors of clinical deterioration the day after hospital admission.


[Spectral studies on nano-sized SO4(2-)/TiO2 photocatalyst prepared by hydrothermal heating the solution of Ti(SO4)2].
In this paper, the nano-sized SO4(2-)/TiO2 solid superacid photocatalysts have been prepared by the method of hydrothermal heating the mixed solution of Ti(S04)2 and urea, heating-decomposing, wet impregnating-calcining. The middle products and the ultimate products have been characterized by XRD, BET, FTIR, DRS and FS. The degradation of rhodamine B by photocatalyst was regarded as a model reaction. The optimum condition of preparing SO4(2-)/TiO2 photocatalysts has been chosen. The results show that hydrothermal treating the solution of Ti(SO4)2 can get the anatase nano-sized TiO2 in the weak alkalinity medium; at 300 degrees C by-products of hydrothermal reaction, such as (NH4)2SO4 and so on, can be decomposed by calcining for 4 hours, avoiding much loss of H2SO4; the most important factors that influence the photocatalystic activities of SO4(2-)/TiO2 are the concentration of SO4(2-) on TiO2 and the calcination time. SO4(2-)/TiO2 photocatalysts have the best photocatalystic activities, up to the level of P-25 photocatalyst, when the concentration of SO4(2-) on TiO2 is 11% and the calcination temperature is 450 degrees C.


Matching of postcontraction perfusion to oxygen consumption across submaximal contraction intensities in exercising humans.
Studying the magnitude and kinetics of blood flow, oxygen extraction, and oxygen consumption at exercise onset and during the recovery from exercise can lead to insights into both the normal control of metabolism and blood flow and the disturbances to these processes in metabolic and cardiovascular diseases. The purpose of this study was to examine the on- and off-kinetics for oxygen delivery, extraction, and consumption as functions of submaximal contraction intensity. Eight healthy subjects performed four 1-min isometric dorsiflexion contractions, with two at 20% MVC and two at 40% MVC. During one contraction at each intensity, relative perfusion changes were measured by using arterial spin labeling, and the deoxyhemoglobin percentage (%HHb) was estimated using the spin- and gradient-echo sequence and a previously published empirical calibration. For the whole group, the mean perfusion did not increase during contraction. The %HHb increased from ∼28 to 38% during contractions of each intensity, with kinetics well described by an exponential function and mean response times (MRTs) of 22.7 and 21.6 s for 20 and 40% MVC, respectively. Following contraction, perfusion increased ∼2.5-fold. The %HHb, oxygen consumption, and perfusion returned to precontraction levels with MRTs of 27.5, 46.4, and 50.0 s, respectively (20% MVC), and 29.2, 75.3, and 86.0 s, respectively (40% MVC). These data demonstrate in human subjects the varied recovery rates of perfusion and oxygen consumption, along with the similar rates of %HHb recovery, across these exercise intensities.


Historical and organizational sources of Córrego do Feijão dam disaster.
Man-made disasters, such as tailings dam failures, raise countless questions. Such is the case of the Vale S.A. dam in Brumadinho, which failed on 25 January 2019. Based on the Accident Analysis and Prevention Model (AAPM), in the present essay we raise some issues deserving of consideration in an in-depth analysis of this disaster. Following AAPM four axes - analysis of the normal operation, barrier analysis, change analysis and conceptual broadening - we sought to contribute to the understanding of the human, technological and organizational dimensions of the disaster. Investigations restricted to technical explanations of the dam failure do not seem to be acceptable. In the present case, analysis should elucidate decision making at several levels of the company management that possibly culminated in normalization of deviance and migration of the system toward accidents. The company's influence on control and overseeing agencies evidences the weaknesses of the Brazilian disaster prevention model.


Intraoperative SD-OCT in macular surgery.
Intraoperative use of spectral-domain optical coherence tomography (SD-OCT) using an OCT-mounted surgical microscope could provide additional information to predict visual outcomes of macular surgery and identify intraoperative changes in the macular anatomy that affect visual recovery. This interventional retrospective case study included 5 eyes of 5 consecutive patients with various macular pathologies. All patients underwent the standard surgery for idiopathic macular holes and epiretinal membranes (ERMs) using the same 25-gauge three-port pars plana technique. If present, the ERM was removed. The dye procedure was then repeated to stain and remove the internal limiting membrane (ILM). OCT images were obtained with the retinal map or cross-section program preincision, post-hyaloid elevation (if applicable), post-residual ERM peel (if applicable), and post-ILM peel. In 3 of 5 patients, the intraoperative scan revealed an increased area of subretinal hyporeflectivity corresponding with a clinically visible "cuff" of retinal detachment. In 2 patients, the SD-OCT scan taken immediately after ERM removal helped find a cleavage plane for the subsequent ILM peeling, which could be further confirmed by a scan taken immediately after it. In 2 cases, the scans identified intraoperative complications that otherwise would not have been noted and allowed the surgeon to modify the next procedures during surgery. Use of this technology in an operative setting can provide greater insight into the changes in retinal anatomy created during macular surgery.


Monoclonal antibody mapping of structural and functional plectin epitopes.
To map structural and functional epitopes of the cytomatrix protein plectin, a set of mAbs was prepared by immunization of mice. Using immunoblot analysis of plectin fragments obtained after limited digestion with various proteases, two groups of mAbs were distinguished. The epitopes of one group (1) were located on a 130-kD terminal segment of the plectin 300-kD polypeptide chain, whereas those of the other group (2) bound within a 40kD segment confined to a central domain of the polypeptide chain. Domains containing the epitopes of group 2 mAbs were shown to include in vitro phosphorylation sites for kinase A, whereas kinase C phosphorylation sites were found on the same terminal segment that contained group 1 mAb epitopes. Rotary shadowing EM of mAb (Fab fragment) -decorated plectin molecules at various states of aggregation, ranging from characteristic dumbbell-shaped single molecules to highly complex multimeric structures, revealed that the epitopes of group 1 as well as those of group 2 mAbs were located on plectin's roughly 200-nm long rod domain interlinking its two globular end domains. Epitopes of group 1 mAbs were localized within a region near the center of the rod, those of group 2 in more peripheral sections near the globular end domains. Solid-phase binding assays carried out in the presence of Fab fragments of mAbs demonstrated an interference of certain group 1 mAbs in the interactions of plectin with vimentin and lamin B. On the other hand, plectin's self-interaction was inhibited mainly by Fab fragments with epitopes in the peripheral rod domain (group 2 mAbs). Together, these results suggested that the molecular binding sites of plectin for vimentin and lamin B, as well as the phosphorylation sites for kinase C, were confined to a defined central section of plectin's rod domain. In addition, they suggest an involvement of peripheral rod sections in plectin self-association.


High-accuracy measurement of the focal length and distortion of optical systems based on interferometry.
A figure measuring interferometer (FMI) method is proposed for high-accuracy measurement of the focal length and distortion of optical systems simultaneously. FMI uses the Zernike coefficients of interference fringe to identify the image point position precisely, and then measures the distance between the image points under the different fields to determine the image height. The field of view can also be accurately obtained by a precise rotating platform. Linear fitting between the field of view and the image height is used to calculate the focal length and distortion. The experimental results indicate that FMI has a relative expanded standard uncertainty of less than 0.01% for focal length and 0.02% for distortion. In brief, the proposed method is feasible for measurement of the focal length and distortion with high accuracy, promising further industrial applications.


Endocrine stress response in rats subjected to singular orbital puncture while under diethyl-ether anaesthesia.
In an attempt to assess possible discomfort in rats subjected to orbital puncture while under diethyl-ether anaesthesia, their endocrine stress response was determined. Concentrations of corticosterone, adrenaline and noradrenaline were measured in plasma obtained via a jugular catheter from rats subjected to diethyl-ether anaesthesia with or without orbital puncture. No statistically significant differences were found between the punctured and non-punctured rats as to peak levels of plasma corticosterone and adrenaline as well as for the times required by the increased concentrations to return to baseline values. The rate by which the plasma noradrenaline level returned to baseline values was somewhat decreased by orbital puncture. Diethyl-ether anaesthesia alone produced a marked endocrine response when compared with handling and novelty stress associated with the induction of anaesthesia. It is concluded that diethyl-ether anaesthesia causes pronounced increases in the plasma levels of the selected stress hormones and that orbital puncture does not amplify this response. It is suggested that diethyl-ether anaesthesia masks any effects of orbital puncture.


Medical education: a neglectful and abusive family system.
Conceptualizing medical education as a family system facilitates recognition of the dynamics within the system which perpetuate many of the problems facing teachers, students, and practicing physicians. Neglectful and abusive families are often characterized by their unrealistic expectations, denial, indirect communication patterns, rigidity, and isolation. The medical education system has similar patterns of behavior that contribute to problems at all levels of the training process and include practicing physicians. The communication patterns within the teaching hospital reinforce trainees' strivings for perfectionism and devalue the contributions of nonphysician staff to the supervision of the physician-in-training. Excluding the potentially healing influence of "outsiders" contributes to the rigidity within the system. Like parents who raise their children as they themselves were raised, each generation teaches as they were taught, and the patterns are loyally perpetuated. Teachers will need to address their own training experiences, acknowledging the dysfunctional behavior patterns learned and the pain those behaviors cause. Then educators can begin to change how they teach and break the cycle of physical neglect and emotional abuse that has been inherited.


Chemical sensing plate with a laser-terahertz monitoring system.
A new type of laser-terahertz emission system for noncontact investigations of chemical solutions has been developed. The system monitors terahertz emission from a sensing plate, which consists of silicon oxide and silicon thin film layers on a sapphire substrate. Sensing of chemical solutions with pH values between 1.68 and 10.01 was demonstrated. The amplitude of the terahertz emission from the sensing plate increased with increasing pH value. This change in the amplitude was caused by a change in the depletion layers of the silicon thin film when protons were adsorbed on the surface of the sensing plate. This study demonstrates that full noncontact monitoring of chemical solutions is possible using the laser-terahertz emission system.


Current epidemiology of intracranial abscesses: a prospective 5 year study.
Intracranial abscesses remain a significant health-care problem in developing countries. In view of this, we undertook a comprehensive study to determine the demographics and bacteriological spectrum of brain abscesses in our hospital. Bacteriological profiles and antibiograms were studied by conventional microbiological methods. Seventy-five patients were admitted with brain abscesses over a 5 year period (2001-2005). There was 9.5% mortality in patients included in this study. The most important factors influencing mortality from intracranial abscess were the age and neurological condition of the patient at the time of admission. Brain abscess could develop at any age but there was a preponderance of males over females. Chronic suppurative otitis media was the most common predisposing factor for temporal lobe infections. Forty-one (54.70%) abscesses were found to be due to pyogenic organisms, 4% due to Mycobacterium tuberculosis and 1.3% were due to Cladophialophora bantiana. The majority of microbial isolates were sensitive to the therapeutic regime adopted in our neurosurgery unit (cefotaxime, gentamicin and metronidazole). Chloramphenicol is another antibiotic with in vitro activity against the isolates.


[Carbohydrate component of Penicillium funiculosum dextranase].
The carbohydrate composition of dextranase from Penicillium funiculosum 15, as well as the composition of products of dextran deep hydrolysis by the enzyme were studied. The products are normally used to stabilize the enzyme during its purification. Using the methods available, it was possible to identify only part of strongly bound (adsorbed) carbohydrates. It was found that dextranase from Pen. funiculosum 15 contained two types of carbohydrates strongly bound with protein: adsorbed and covalently bound carbohydrates. A procedure allowing a complete separation of adsorbed carbohydrates was developed. The procedure is based on the use of stabilizing additives of readily separable carbohydrates. The enzyme, which is shown by polyacrylamide gel electrophoresis in the presence of Na-dodecyl sulfate and beta-mercaptoethanol to be homogeneous, consists of 313 amino acid residues, 3 glucosamine residues and residues of mannose, galactose and fucose in the ratio 6:2:1.


Noninvasive diagnosis of deep venous thrombosis.
Eighty-five patients suspected of having lower-extremity deep venous thrombosis (DVT) participated in a prospective study to test the diagnostic accuracy of four noninvasive techniques: Doppler ultrasonic flow study, electrical impedance plethysmography, the serial dilution protamine sulfate test, and an extensive physical examination. Ascending radiocontrast phlebography was the diagnostic standard of reference. We found that (1) when both Doppler and impedance examinations were positive, the diagnosis of DVT could be considered virtually certain; (2) impedance and Doppler examinations, when used in combination, were reliable screening tests capable of establishing or excluding the presence of thigh DVT; (3) physical examination and the serial dilution protamine sulfate test were unreliable screening techniques for DVT; (4) techniques other than the noninvasive methods investigated were needed to reliably detect or to exclude popliteal and call DVT.


A novel approach to implant screw-retained restorations: adhesive combination between zirconia frameworks and monolithic lithium disilicate.
The use of zirconia is an esthetic alternative to metal for implant-supported frameworks, and it has increased primarily for its high biocompatibility, low bacterial surface adhesion, high flexural strength and high mechanical features. The zirconia frameworks in fixed prosthetic restorations that are supported by implants is commonly covered with hand-layered overlay porcelain. This technical procedure is highly esthetic but it can cause some complications, such as porcelain fractures. The purpose of this article is to introduce an innovative approach to create an esthetic fixed ceramic implant restoration to minimize and facilitate the repair of the mechanical complications, by combining the adhesive-cementation of lithium disilicate full coverage restorations on implant screw-retained zirconia frameworks.


The Analysis of Three-Body Contact Temperature under the Different Third Particle Size, Density, and Value of Friction.
Recently, many studies have investigated the friction, wear, and temperature characteristics of the interface between two relative movements. Such analyses often set the coefficient of friction as a fixed value and are analyzed in cases of two-body contact; however, the interface is often a three-body contact and the coefficient of friction varies depending on the operating conditions. This is a significant error in the analysis of contact characteristics, therefore, in this study, the actual interface and the change of the coefficient of friction were analyzed based on three-body micro-contact theory where the contact temperature was also analyzed and the difference between the generally assumed values were compared. The results showed that under three-body contact, the coefficient of total friction increased with an increase in particle size; and at a different particle size and area density of particles, the surface contact temperature increased with the plasticity index and load increases, and the particle contact temperature increased with the increasing particle size. The surface temperature rise was mainly affected by the ratio of the average temperature between surface 1 and surface 2 to the multiplication between the 100th root of the area density of particles and the square root of the equivalent surface roughness (Ts1s2_ave*/ηa0.01σ0.5) and the ratio of the 10th root of the mean particle diameter to the 100th root of the equivalent surface roughness (xa0.1/σ0.001). Particle temperature was mainly affected by the ratio of the 10th root of the mean particle diameter to the 100th root of the equivalent surface roughness (xa0.1/σ0.001) and the area density of particles ηa. Our study indicated that when the contact of surface with surface and the contact of the particles with the surface, the resulting heat balance was assigned to the particles and the surface in a three-body contact situation. Under this contact behavior, it could avoid a too high a rise in micro-contact temperature to achieve the material failure temperature.


Identification of distinct steps during tubule formation by the movement protein of Cowpea mosaic virus.
The movement protein (MP) of Cowpea mosaic virus (CPMV) forms tubules through plasmodesmata in infected plants thus enabling virus particles to move from cell to cell. Localization studies of mutant MPs fused to GFP in protoplasts and plants identified several functional domains within the MP that are involved in distinct steps during tubule formation. Coinoculation experiments and the observation that one of the C-terminal deletion mutants accumulated uniformly in the plasma membrane suggest that dimeric or multimeric MP is first targeted to the plasma membrane. At the plasma membrane the MP quickly accumulates in peripheral punctuate spots, from which tubule formation is initiated. One of the mutant MPs formed tubules containing virus particles on protoplasts, but could not support cell-to-cell movement in plants. The observations that this mutant MP accumulated to a higher level in the cell than wt MP and did not accumulate in the cell wall opposite infected cells suggest that breakdown or disassembly of tubules in neighbouring, uninfected cells is required for cell-to-cell movement.


Locally adaptive activation functions with slope recovery for deep and physics-informed neural networks.
We propose two approaches of locally adaptive activation functions namely, layer-wise and neuron-wise locally adaptive activation functions, which improve the performance of deep and physics-informed neural networks. The local adaptation of activation function is achieved by introducing a scalable parameter in each layer (layer-wise) and for every neuron (neuron-wise) separately, and then optimizing it using a variant of stochastic gradient descent algorithm. In order to further increase the training speed, an activation slope-based slope recovery term is added in the loss function, which further accelerates convergence, thereby reducing the training cost. On the theoretical side, we prove that in the proposed method, the gradient descent algorithms are not attracted to sub-optimal critical points or local minima under practical conditions on the initialization and learning rate, and that the gradient dynamics of the proposed method is not achievable by base methods with any (adaptive) learning rates. We further show that the adaptive activation methods accelerate the convergence by implicitly multiplying conditioning matrices to the gradient of the base method without any explicit computation of the conditioning matrix and the matrix-vector product. The different adaptive activation functions are shown to induce different implicit conditioning matrices. Furthermore, the proposed methods with the slope recovery are shown to accelerate the training process.


Hyperhexosemia-Induced Retinal Vascular Pathology in a Novel Primate Model of Diabetic Retinopathy.
The paucity of animal models exhibiting full pathology of diabetic retinopathy (DR) has impeded understanding of the pathogenesis of DR and the development of therapeutic interventions. Here, we investigated whether hyperhexosemic marmosets (Callithrix jacchus) develop characteristic retinal vascular lesions including macular edema (ME), a leading cause of vision loss in DR. Marmosets maintained on 30% galactose (gal)-rich diet for 2 years were monitored for retinal vascular permeability, development of ME, and morphological characteristics including acellular capillaries (AC) and pericyte loss (PL), vessel tortuosity, and capillary basement membrane (BM) thickness. Excess vascular permeability, increased number of AC and PL, vascular BM thickening, and increased vessel tortuosity were observed in the retinas of gal-fed marmosets. Optical coherence tomography (OCT) images revealed significant thickening of the retinal foveal and the juxtafoveal area, and histological analysis showed incipient microaneurysms in retinas of gal-fed marmosets. Findings from this study indicate that hyperhexosemia can trigger retinal vascular changes similar to those seen in human DR including ME and microaneurysms. The striking similarities between the marmoset retina and the human retina, and the exceptionally small size of the monkey, offer significant advantages to this primate model of DR.


Iron catalyzed conversion of NO into nitrosonium (NO+) and nitroxyl (HNO/NO-) species.
The conversion of NO into its congeners, nitrosonium (NO+) and nitroxyl (HNO/NO-) species, has important consequences in NO metabolism. Dinitrosyl iron complex (DNIC) combined with thiol ligands was shown to catalyze the conversion of NO into NO+, resulting in the synthesis of S-nitrosothiols (RSNO) both in vitro and in vivo. The formation mechanism of DNIC was proposed to involve the intermediate release of nitroxyl. Since the detection of hydroxylamine (as the product of a rapid reaction of HNO/NO- with thiols) is taken as the evidence for nitroxyl generation, we examined the formation of hydroxylamine, RSNO, and nitrite (the product of a rapid reaction of NO+ with water) in neutral solutions containing iron ions and thiols exposed to NO under anaerobic conditions. Hydroxylamine was detected in NO treated solutions of iron ions in the presence of cysteine, but not glutathione (GSH). The addition of urate, a major "free" iron-binding agent in humans, to solutions of GSH and iron ions, and the subsequent treatment of these solutions with NO increased the synthesis of GSNO and resulted in the formation of hydroxylamine. This caused a loss of urate and yielded a novel nitrosative/nitration product. GSH attenuated the urate decomposition to such a degree that it could be reflected as the function of GSH:urate. Results described here contribute to the understanding of the role of iron ions in catalyzing the conversion of NO into HNO/NO- and point to the role of uric acid not previously described.


Strong ultrasonic microwaves in ferroelectric ceramics.
It is well known that ferroelectric materials have piezoelectric properties which allow the transformation of electrical signals into mechanical signals and vice versa. The transducer action normally is restricted to frequencies up to the mechanical resonance frequency of the sample. There are, however, two mechanisms which allow transducer action in ferroelectric ceramics at much higher frequencies: one is the normal piezoelectric effect in a ferroelectric ceramic in which the crystallites have periodic domain structures, the other is a domain wall effect in which ferroelastic domain walls in a periodic domain structure are powerful shear wave emitters. Both mechanisms give rise to extensive dielectric losses in ceramics at microwave frequencies.