Quantification of oxygen-induced retinopathy in the mouse.
To describe a quantifiable model of vascular proliferation in oxygen-induced retinopathy (OIR) in the mouse. Neonate Quackenbush mice were subjected to high-ambient oxygen (approximately 95%) for the first 5 days after birth, effecting a total inhibition of retinal vascular growth. Animals then were returned to room air, and the rates of subsequent vascular development in the plane of the retina and estimates of retinal capillary density were measured from flatmounts of ink-perfused eyes. Observations were confirmed with fluorescein isothiocyanate-lectin labeling of the peripheral vasculature. Abnormal growth of vascular sprouts into the vitreous was recorded from cross-sections. Observations in OIR were compared against those of age-matched control animals. The slower rate of retinal revascularization in OIR mice was quantified and compared against the normal rate. Lectin-binding studies confirmed the reliability of ink preparations. The number of vitreous sprouts in OIR peaked 8 to 10 days after animals were returned to room air (13 to 15 postnatal days). Sprouts then regressed, to disappear by postnatal day 20. In all respects, bar a slightly lower peripheral capillary density, the normal retinal vascular pattern was achieved in OIR within 15 days of exposure to room air (as opposed to the 10 days required in control mice). The protocol described for quantifying retinal proliferation in mouse OIR is reproduced readily, and the data recorded here will allow the effectiveness of subsequent treatments that may affect retinal vascular growth to be evaluated better.


Molecular characterization of a long range haplotype affecting protein yield and mastitis susceptibility in Norwegian Red cattle.
Previous fine mapping studies in Norwegian Red cattle (NRC) in the region 86-90.4 Mb on Bos taurus chromosome 6 (BTA6) has revealed a quantitative trait locus (QTL) for protein yield (PY) around 88 Mb and a QTL for clinical mastitis (CM) around 90 Mb. The close proximity of these QTLs may partly explain the unfavorable genetic correlation between these two traits in NRC. A long range haplotype covering this region was introduced into the NRC population through the importation of a Holstein-Friesian bull (1606 Frasse) from Sweden in the 1970s. It has been suggested that this haplotype has a favorable effect on milk protein content but an unfavorable effect on mastitis susceptibility. Selective breeding for milk production traits is likely to have increased the frequency of this haplotype in the NRC population. Association mapping for PY and CM in NRC was performed using genotypes from 556 SNPs throughout the region 86-97 Mb on BTA6 and daughter-yield-deviations (DYDs) from 2601 bulls made available from the Norwegian dairy herd recording system. Highest test scores for PY were found for single-nucleotide polymorphisms (SNPs) within and surrounding the genes CSN2 and CSN1S2, coding for the β-casein and α(S2)-casein proteins. High coverage re-sequencing by high throughput sequencing technology enabled molecular characterization of a long range haplotype from 1606 Frasse encompassing these two genes. Haplotype analysis of a large number of descendants from this bull indicated that the haplotype was not markedly disrupted by recombination in this region. The haplotype was associated with both increased milk protein content and increased susceptibility to mastitis, which might explain parts of the observed genetic correlation between PY and CM in NRC. Plausible causal polymorphisms affecting PY were detected in the promoter region and in the 5'-flanking UTR of CSN1S2. These polymorphisms could affect transcription or translation of CSN1S2 and thereby affect the amount of α(S2)-casein in milk. Highest test scores for CM were found in the region 89-91 Mb on BTA6, very close to a cluster of genes coding for CXC chemokines. Expression levels of some of these CXC chemokines have previously been shown to increase in bovine mammary gland cell lines after exposure to bacterial cell wall components. Molecular characterization of the long range haplotype from the Holstein-Friesian bull 1606 Frasse, imported into NRC in the 1970s, revealed polymorphisms that could affect transcription or translation of the casein gene CSN1S2. Sires with this haplotype had daughters with significantly elevated milk protein content and selection for milk production traits is likely to have increased the frequency of this haplotype in the NRC population. The haplotype was also associated with increased mastitis susceptibility, which might explain parts of the genetic correlation between PY and CM in NRC.


RECK expression in pancreatic cancer: its correlation with lower invasiveness and better prognosis.
The reversion-inducing cysteine-rich protein with Kazal motifs (RECK) gene was initially isolated as a transformation suppressor gene. The RECK gene is expressed widely in normal organs but is undetectable in many tumor-derived cell lines. When artificially expressed in such cell lines, RECK negatively regulates at least matrix metalloprotease (MMP)-9, MMP-2, and MT1-MMP activation and suppresses the invasive and metastatic potentials of these cells. Clinical relevance of these observations, however, is yet to be established. The aim of this study was to examine RECK expression in pancreatic cancer, where intensive invasiveness and metastasis are frequently observed, and investigate its clinical significance. We also analyzed the correlation between RECK expression and MMP activation. (a) RECK expression in surgically resected tissue samples of invasive ductal carcinomas of the pancreas (n = 50) was examined immunohistochemically, and its correlation with clinicopathological factors was analyzed; and (b) gelatin zymography was used for the detection of latent and activated forms of MMP-2 and MMP-9 in some of the tissue samples (n = 33). The gelatinase activity was quantified by densitometory, and the ratio of intensity of the active MMP-2 band to the total intensity of the pro- and active MMP-2 bands was evaluated as an indicator of MMP-2 activation. The MMP-9 activation was also studied. Among the 50 ductal carcinoma samples, 26 (52%) were stained positive for RECK. In the normal pancreas, both acinar and beta cells were stained positive, but ductal cells did not. Tumors with positive RECK staining were significantly less invasive as compared with RECK-negative tumors (P = 0.0438). Importantly, patients who had tumors with high RECK expression showed significantly better prognosis than those who had RECK-negative tumors (P = 0.0463, by Log-rank test). Zymographic analysis indicated significant inverse correlation between the level of RECK expression and extent of MMP-2 activation (P = 0.0374). Our findings support the hypothesis that the RECK protein has negative effects on the invasiveness of pancreatic cancer by inhibiting MMP-2 activation and suggest the potential value of RECK as a prognostic molecular marker for pancreatic cancer.


[Ventricular septal defect repair in children during first year of life in a public hospital].
Ventricular septal defect (VSD) is the most common congenital heart disease; primary surgical closure is the usual strategy for repairing it. Our objective is to describe results of surgical repair in children under 1 year of age and analyze risk factors for morbidity and mortality. Retrospective study; all patients with VSD repaired between 2004 and 2011 were included. Demographic, surgical procedure and postoperative variables were recorded: age, weight, genetic syndrome, type of VSD, length of stay, complications and outcome. Risk factors of mortality and morbidity: age < 6 m, weight < 3 kg, Down, malnutrition and respiratory infection prior syndrome were analyzed. 256 patients, age 5.3 months (21d-1y), weight 4.75 kg (2.2 to 13), 32% with Down syndrome and 17.5% with preoperative mechanical ventilation were operated. Perimembranous VSD was the most frequent type (62%). 28% experienced complications and 7% required reoperation for bleeding, infection or defect. The median hospital stay was 6 days (1-185). Postoperative 30 days mortality was 3%. Age < 6 m, weight < 3 kg, malnutrition and prior respiratory viral infection were associated with prolonged hospitalization, but no risk factors for mortality were identified. The primary surgical closure of the VSD is a procedure with satisfactory results at our institution.


Identification of a cluster of residues in transmembrane segment 6 of domain III of the cockroach sodium channel essential for the action of pyrethroid insecticides.
A phenylalanine residue (Phe1519) in the sixth transmembrane segment of domain III (IIIS6) of the cockroach BgNa(v) sodium channel is required for the binding and action of pyrethroids. However, whether or not other residues in IIIS6 participate in the action of pyrethroids remains to be determined. In the present study, we conducted a systematic analysis of 20 residues in IIIS6 of the BgNa(v) channel using alanine-scanning mutagenesis. Our results show that alanine substitutions of four residues, Ile1514, Gly1516, Phe1518 and Asn1522, altered sodium channel sensitivity to pyrethroid insecticides. Whereas the G1516A, F1518A and N1522A substitutions diminished sodium channel sensitivity to all seven pyrethroids examined, including four type I (lacking the alpha-cyano group at the phenoxybenzyl alcohol) and three type II (containing the alpha-cyano group) pyrethroids, the I1514A substitution enhanced sodium channel sensitivity to four type I and type II pyrethroids that contain the phenoxybenzyl alcohol only. We also show that alanine/lysine substitutions of Leu1521 and Ser1517 affected the action of BTX (batrachotoxin), but not pyrethroids. In the Kv1.2-based homology model of the open sodium channel, side chains of Ile1514, Phe1518 and Asn1522 are exposed towards helix IIS5 and linker IIS4-IIS5, which contain previously identified pyrethroid-interacting residues, whereas Ser1517 and Leu1521 face the inner pore where the BTX receptor is located. Thus the present study provides further evidence for structural models in which pyrethroids bind to the lipid-exposed interface formed by helices IIIS6, IIS5 and linker helix IIS4-IIS5, whereas BTX binds to the pore-exposed side of the IIIS6 helix.


Stat3 activation regulates the expression of vascular endothelial growth factor and human pancreatic cancer angiogenesis and metastasis.
Expression of vascular endothelial growth factor (VEGF), a key angiogenic protein, has been linked with pancreatic cancer progression. However, the molecular basis for VEGF overexpression remains unclear. Immunohistochemical studies have indicated that VEGF overexpression coincides with elevated Stat3 activation in human pancreatic cancer specimens. In our study, more than 80% of the human pancreatic cancer cell lines used exhibited constitutively activated Stat3, with Stat3 activation correlated with the VEGF expression level. Blockade of activated Stat3 via ectopic expression of dominant-negative Stat3 significantly suppressed VEGF expression, angiogenesis, tumor growth, and metastasis in vivo. Furthermore, constitutively activated Stat3 directly activated the VEGF promoter, whereas dominant-negative Stat3 inhibited the VEGF promoter. A putative Stat3-responsive element on the VEGF promoter was identified using a protein-DNA binding assay and confirmed using a promoter mutagenesis assay. These results indicate that Stat3 directly regulates VEGF expression and hence angiogenesis, growth, and metastasis of human pancreatic cancer, suggesting that Stat3 signaling may be targeted for treatment of pancreatic cancer.


MRI quantification of muscle activity after volitional exercise and neuromuscular electrical stimulation.
The efficacy, and even the depth, of muscle stimulation during surface electrode neuromuscular electrical stimulation (NMES) is a matter of debate. This study addresses these issues by assessing the utility of a magnetic resonance imaging (MRI) technique in localizing and quantitating changes in the nature of MRI signals in the quadriceps muscle after volitional exercise and NMES. Volitional isometric and NMES-evoked quadriceps muscle activity was evaluated in two controlled trials. In the first, isometric quadriceps strength was determined during NMES and maximal volitional isometric exercise in six healthy men. In the second, changes in the ratio of MRI T2 signal intensities before and after volitional isometric exercise and NMES were used to quantitate MRI signal changes associated with muscle activation in 12 additional healthy men. MRI clearly detected quadriceps muscle tissue activation after both volitional and stimulated contractions, even though the NMES knee extension torque was only 23.5% that of maximal volitional isometric exercise. In particular, the T2 intensity ratios increased 26.5% +/- 17.3% (mean +/- standard deviation) after volitional exercise and 12.9% +/- 12.8% after NMES. This pattern of volitional isometric exercise, producing larger T2 intensity ratio values than NMES, was present in both deep and superficial layers and throughout the quadriceps muscle. Although volitional muscle contractions were several times stronger than those induced by NMES in this study, our findings support the idea that MRI can provide a noninvasive way to quantitate and localize volitional and electrically stimulated muscle activation.


Ureterolithiasis: Management in an environment with limited facilities.
In the past 2-3 decades, there has been a dramatic development in the techniques of stone removal. This study highlights the management of symptomatic ureteral stones in an environment without such facilities. Sixty-nine patients, comprising 53 (76.8%) males and 16 (23.2%) females, diagnosed of symptomatic ureteric calculi within the study period in two tertiary health institutions were included in the study. Thorough history taking and physical examinations were performed. Extensive laboratory investigations using blood and urine specimens were carried out. Imaging studies, ultrasonography, intravenous urography, and computerized tomographic scan were used to locate the position and size of the calculi. Forty-six (66.7%) patients presented with excruciating flank ureteric colic radiating to the groin in 16 (23.2%) patients and hematuria in 62 (89.9%) patients. Bilateral ureteric calculi occurred in 3 (4.3%) patients. Eleven (15%) stones passed spontaneously. 33 (47.8%) patients had uneventful open surgery. The stones were mixed in nature. Management of ureteric stones in our environment is affected by delay in presentation, low level of awareness of urinary stone disease, lack of modern endourological equipment, and paucity of urological surgeons. Finally, medical treatment should be explored for stones below 10 mm in size.


Zernike expansion of derivatives and Laplacians of the Zernike circle polynomials.
The partial derivatives and Laplacians of the Zernike circle polynomials occur in various places in the literature on computational optics. In a number of cases, the expansion of these derivatives and Laplacians in the circle polynomials are required. For the first-order partial derivatives, analytic results are scattered in the literature. Results start as early as 1942 in Nijboer's thesis and continue until present day, with some emphasis on recursive computation schemes. A brief historic account of these results is given in the present paper. By choosing the unnormalized version of the circle polynomials, with exponential rather than trigonometric azimuthal dependence, and by a proper combination of the two partial derivatives, a concise form of the expressions emerges. This form is appropriate for the formulation and solution of a model wavefront sensing problem of reconstructing a wavefront on the level of its expansion coefficients from (measurements of the expansion coefficients of) the partial derivatives. It turns out that the least-squares estimation problem arising here decouples per azimuthal order m, and per m the generalized inverse solution assumes a concise analytic form so that singular value decompositions are avoided. The preferred version of the circle polynomials, with proper combination of the partial derivatives, also leads to a concise analytic result for the Zernike expansion of the Laplacian of the circle polynomials. From these expansions, the properties of the Laplacian as a mapping from the space of circle polynomials of maximal degree N, as required in the study of the Neumann problem associated with the transport-of-intensity equation, can be read off within a single glance. Furthermore, the inverse of the Laplacian on this space is shown to have a concise analytic form.


Neonatal induction of tolerance to skeletal tissue allografts without immunosuppression.
Vascularized allogeneic skeletal tissue transplantation without the need for host immunosuppression would increase reconstructive options for treating congenital and acquired defects. Because the immune system of a fetus or neonate is immature, it may be possible to induce tolerance to allogeneic skeletal tissues by alloantigen injection during this permissive period. Within 12 hours after birth, 17 neonatal Lewis rats were injected through the superficial temporal vein with 3.5 to 5 million Brown Norway bone marrow cells in 0.1 ml normal saline. Ten weeks after the injection, peripheral blood from the Lewis rats was analyzed for the presence of Brown Norway cells to determine hemopoietic chimerism. The Lewis rats then received a heterotopic, vascularized limb tissue transplant (consisting of the knee, the distal femur, the proximal tibia, and the surrounding muscle on a femoral vascular pedicle) from Brown Norway rat donors to determine their tolerance to the allogeneic tissue. A positive control group (n = 6) consisted of syngeneic transplants from Lewis rats into naive Lewis rats to demonstrate survival of transplants. A negative control group (n = 6) consisted of Brown Norway transplants into naive Lewis rats not receiving bone marrow or other immunosuppressive treatment. The animals were assessed for transplant viability 30 days after transplantation using histologic and bone fluorochrome analysis. All the syngeneic controls (Lewis to Lewis) remained viable throughout the experiment, whereas all the Brown Norway to Lewis controls had rejected. Ten of the 17 allografts transplanted into bone marrow recipients were viable at 30 days, with profuse bleeding from the ends of the bone graft and the surrounding graft muscle. The percent of chimerism correlated with survival, with 3.31 percent (SD = 1.9) of peripheral blood, Brown Norway chimerism present in the prolonged survival groups and 0.75 percent (SD = 0.5) of Brown Norway chimerism in the rejected graft group. This study demonstrated prolonged survival of allogeneic skeletal tissue without immunosuppression after early neonatal injection of allogeneic bone marrow in a rat model.


Deficiency of multidrug resistance 2 contributes to cell transformation through oxidative stress.
Multidrug resistance 2 (Mdr2), also called adenosine triphosphate-binding cassette B4 (ABCB4), is the transporter of phosphatidylcholine (PC) at the canalicular membrane of mouse hepatocytes, which plays an essential role for bile formation. Mutations in human homologue MDR3 are associated with several liver diseases. Knockout of Mdr2 results in hepatic inflammation, liver fibrosis and hepatocellular carcinoma (HCC). Whereas the pathogenesis in Mdr2 (-/-) mice has been largely attributed to the toxicity of bile acids due to the absence of PC in the bile, the question of whether Mdr2 deficiency per se perturbs biological functions in the cell has been poorly addressed. As Mdr2 is expressed in many cell types, we used mouse embryonic fibroblasts (MEF) derived from Mdr2 (-/-) embryos to show that deficiency of Mdr2 increases reactive oxygen species accumulation, lipid peroxidation and DNA damage. We found that Mdr2 (-/-) MEFs undergo spontaneous transformation and that Mdr2 (-/-) mice are more susceptible to chemical carcinogen-induced intestinal tumorigenesis. Microarray analysis in Mdr2-/- MEFs and cap analysis of gene expression in Mdr2 (-/-) HCCs revealed extensively deregulated genes involved in oxidation reduction, fatty acid metabolism and lipid biosynthesis. Our findings imply a close link between Mdr2 (-/-) -associated tumorigenesis and perturbation of these biological processes and suggest potential extrahepatic functions of Mdr2/MDR3.


Presumptive identification of streptococci by pyrrolidonyl-beta-naphthylamide (PYR) test.
Group A streptococci and enterococci can be differentiated from other streptococci on the basis of their ability to cleave L-pyrrolidonyl-beta-naphthylamide. In the present study, the L-pyrrolidonyl-beta-naphthylamide (PYR) test, pigment medium and bile esculin medium have been used to presumptively identify the streptococci. In total, 114 strains of group A streptococci, 350 strains of non-group A streptococci, 202 strains of enterococci and 197 strains of non-enterococci have been tested. The results of the present investigation show that sensitivities of different test methods are: PYR broth, 99.08%; Murex PYR, 98.48%; bacitracin, 95%; bacitracin and sulfamethoxazole/trimethoprim (SXT), 95%; pigment medium, 99.23%; bile esculin medium, 99.26%. Additionally, specificities of various tests are: PYR broth and Murex PYR, 99.82%; bacitracin, 90.90%; bacitracin and SXT, 98.87%; pigment medium and bile esculin medium, 100%, respectively. PYR test has been observed to be very easy to use and may hence be considered as a rapid, reliable and cost-effective method for presumptive identification of group A streptococci and enterococci in the clinical laboratory.


Revealing life-history traits by contrasting genetic estimations with predictions of effective population size.
Effective population size, a central concept in conservation biology, is now routinely estimated from genetic surveys and can also be theoretically predicted from demographic, life-history, and mating-system data. By evaluating the consistency of theoretical predictions with empirically estimated effective size, insights can be gained regarding life-history characteristics and the relative impact of different life-history traits on genetic drift. These insights can be used to design and inform management strategies aimed at increasing effective population size. We demonstrated this approach by addressing the conservation of a reintroduced population of Asiatic wild ass (Equus hemionus). We estimated the variance effective size (Nev ) from genetic data (N ev =24.3) and formulated predictions for the impacts on Nev of demography, polygyny, female variance in lifetime reproductive success (RS), and heritability of female RS. By contrasting the genetic estimation with theoretical predictions, we found that polygyny was the strongest factor affecting genetic drift because only when accounting for polygyny were predictions consistent with the genetically measured Nev . The comparison of effective-size estimation and predictions indicated that 10.6% of the males mated per generation when heritability of female RS was unaccounted for (polygyny responsible for 81% decrease in Nev ) and 19.5% mated when female RS was accounted for (polygyny responsible for 67% decrease in Nev ). Heritability of female RS also affected Nev ; hf2=0.91 (heritability responsible for 41% decrease in Nev ). The low effective size is of concern, and we suggest that management actions focus on factors identified as strongly affecting Nev, namely, increasing the availability of artificial water sources to increase number of dominant males contributing to the gene pool. This approach, evaluating life-history hypotheses in light of their impact on effective population size, and contrasting predictions with genetic measurements, is a general, applicable strategy that can be used to inform conservation practice.


Racial/ethnic disparities in infant mortality.
This study examines predictors of neonatal and postneonatal mortality among infants born to black, white, and Hispanic women. Linked birth/infant death records from North Carolina for the period 1999-2007 were the source of data. Logistic regression models were constructed to estimate the effect of maternal and infant characteristics on neonatal (<28 days) and postneonatal (28-364 days) mortality. Analyses revealed no racial/ethnic differences in neonatal mortality, but increased risk among black infants for postneonatal death. Inadequate prenatal care was associated with an increased risk for neonatal mortality across all three racial/ethnic groups. Younger maternal age and lower educational levels were associated with postneonatal deaths for black and white women, but not Hispanic. A previous child loss, being unmarried and labor/delivery complications increased the risk for postneonatal mortality only among white women. Black infants had higher rates of death attributed to sudden infant death syndrome (SIDS), infections, low birth weight (LBW)/prematurity, respiratory conditions, and injuries. Early initiation of prenatal care, access to risk-appropriate obstetric and neonatal services, and participation in intervention programs that support parenting of LBW/preterm infants throughout the first year of life are likely to yield the greatest impact in reducing infant mortality.


Stroke in Indigenous Africans, African Americans, and European Americans: Interplay of Racial and Geographic Factors.
The relative contributions of racial and geographic factors to higher risk of stroke in people of African ancestry have not been unraveled. We compared stroke type and contributions of vascular risk factors among indigenous Africans (IA), African Americans (AA), and European Americans (EA). SIREN (Stroke Investigative Research and Educational Network) is a large multinational case-control study in West Africa-the ancestral home of 71% AA-whereas REGARDS (Reasons for Geographic and Racial Differences in Stroke) is a cohort study including AA and EA in the United States. Using harmonized assessments and standard definitions, we compared data on stroke type and established risk factors for stroke in acute stroke cases aged ≥55 years in both studies. There were 811 IA, 452 AA, and 665 EA stroke subjects, with mean age of 68.0±9.3, 73.0±8.3, and 76.0±8.3 years, respectively (P<0.0001). Hemorrhagic stroke was more frequent among IA (27%) compared with AA (8%) and EA (5.4%; P<0.001). Lacunar strokes were more prevalent in IA (47.1%), followed by AA (35.1%) and then EA (21.0%; P<0.0001). The frequency of hypertension in decreasing order was IA (92.8%), followed by AA (82.5%) and then EA (64.2%; P<0.0001) and similarly for diabetes mellitus IA (38.3%), AA (36.8%), and EA (21.0%; P<0.0001). Premorbid sedentary lifestyle was similar in AA (37.7%) and EA (34.0%) but lower frequency in IA (8.0%). Environmental risk factors such as sedentary lifestyle may contribute to the higher proportion of ischemic stroke in AA compared with IA, whereas racial factors may contribute to the higher proportion of hypertension and diabetes mellitus among stroke subjects of African ancestry.


Moderate doping leads to high performance of semiconductor/insulator polymer blend transistors.
Polymer transistors are being intensively developed for next-generation flexible electronics. Blends comprising a small amount of semiconducting polymer mixed into an insulating polymer matrix have simultaneously shown superior performance and environmental stability in organic field-effect transistors compared with the neat semiconductor. Here we show that such blends actually perform very poorly in the undoped state, and that mobility and on/off ratio are improved dramatically upon moderate doping. Structural investigations show that these blend layers feature nanometre-scale semiconductor domains and a vertical composition gradient. This particular morphology enables a quasi three-dimensional spatial distribution of semiconductor pathways within the insulating matrix, in which charge accumulation and depletion via a gate bias is substantially different from neat semiconductor, and where high on-current and low off-current are simultaneously realized in the stable doped state. Adding only 5 wt% of a semiconducting polymer to a polystyrene matrix, we realized an environmentally stable inverter with gain up to 60.


A difficult phase for introns-early. Molecular evolution.
Close analysis of intron phase - the position of introns within codons - is claimed to provide novel evidence supporting the view that introns predate the divergence of bacteria and eukaryotes and, via 'exon shuffling', played a crucial role in protein evolution. But just how compelling is this evidence?


Structure of Te-rich Te-Ge-X (X = I, Se, Ga) glasses.
The structure of glassy Te(78)Ge(11)Ga(11), Te(79)Ge(16)Ga(5), Te(70)Ge(20)Se(10) and Te(73)Ge(20)I(7)--promising materials for far infrared applications--was investigated by means of x-ray and neutron diffraction as well as extended x-ray absorption fine structure measurements at various edges. Experimental data sets were fitted simultaneously in the framework of the reverse Monte Carlo simulation technique. Short range order in Te(85)Ge(15) was reinvestigated by fitting a new x-ray diffraction measurement together with available neutron diffraction and extended x-ray absorption fine structure data. It was found that Te(85)Ge(15) consists mostly of GeTe(4) structural units linked together directly or via bridging Te atoms. Te is predominantly twofold coordinated in Te(85)Ge(15), Te(70)Ge(20)Se(10) and Te(73)Ge(20)I(7) while in Te(78)Ge(11)Ga(11) and Te(79)Ge(16)Ga(5) the Te coordination number is significantly higher than 2. The Te-Te bond length is 2.80 ± 0.02 Å in Te(78)Ge(11)Ga(11) while it is as short as 2.70 ± 0.02 Å and 2.73 ± 0.02 Å in Te(73)Ge(20)I(7) and Te(70)Ge(20)Se(10), respectively. Our results show that the strengths of GeTe(4) (GeTe(3)I, GeTe(3)Se) 'units' are very similar in all glasses investigated but the connection between these units depends on the third component. Differences in the Te coordination number suggest that unlike Se or I, Ga does not build into the Ge-Te covalent network. Instead, it forms a covalent bond with the non-bonding p electrons of Te, which results in an increase in the average Te coordination number.


Proteomic profiling of antisense-induced exon skipping reveals reversal of pathobiochemical abnormalities in dystrophic mdx diaphragm.
The disintegration of the dystrophin-glycoprotein complex represents the initial pathobiochemical insult in Duchenne muscular dystrophy. However, secondary changes in signalling, energy metabolism and ion homeostasis are probably the main factors that eventually cause progressive muscle wasting. Thus, for the proper evaluation of novel therapeutic approaches, it is essential to analyse the reversal of both primary and secondary abnormalities in treated muscles. Antisense oligomer-mediated exon skipping promises functional restoration of the primary deficiency in dystrophin. In this study, an established phosphorodiamidate morpholino oligomer coupled to a cell-penetrating peptide was employed for the specific removal of exon 23 in the mutated mouse dystrophin gene transcript. Using DIGE analysis, we could show the reversal of secondary pathobiochemical abnormalities in the dystrophic diaphragm following exon-23 skipping. In analogy to the restoration of dystrophin, beta-dystroglycan and neuronal nitric oxide synthase, the muscular dystrophy-associated differential expression of calsequestrin, adenylate kinase, aldolase, mitochondrial creatine kinase and cvHsp was reversed in treated muscle fibres. Hence, the re-establishment of Dp427 coded by the transcript missing exon 23 has counter-acted dystrophic alterations in Ca2+-handling, nucleotide metabolism, bioenergetic pathways and cellular stress response. This clearly establishes the exon-skipping approach as a realistic treatment strategy for diminishing diverse downstream alterations in dystrophinopathy.


Nationalism and social welfare in the post-Soviet context.
This paper offers hypotheses on the role that state social welfare measures can play in reflecting nationalism and in aggravating interethnic tensions. Social welfare is often overlooked in theoretical literature on nationalism, because of the widespread assumption that the welfare state promotes social cohesion. However, social welfare systems may face contradictions between the goal of promoting universal access to all citizens on the one hand, and social pressures to recognize particular groups in distinct ways on the other. Examples from the post-Soviet context (particularly Russia) are offered to illustrate the ways in which social welfare issues may be perceived as having ethnic connotations.


Auditory brainstem response as a diagnostic tool for patients suffering from schizophrenia, attention deficit hyperactivity disorder, and bipolar disorder: protocol.
Psychiatric disorders, such as schizophrenia, attention deficit hyperactivity disorder (ADHD), and bipolar disorder, may sometimes be difficult to diagnose. There is a great need for a valid and reliable diagnostic tool to aid clinicians in arriving at the diagnoses in a timely and accurate manner. Prior studies have suggested that patients suffering from schizophrenia and ADHD may process certain sound stimuli in the brainstem in an unusual manner. When these patient groups have been examined with the electrophysiological method of brainstem audiometry, some studies have found illness-specific aberrations. Such aberrations may also exist for patients suffering from bipolar disorder. In this study, we will examine whether the method of brainstem audiometry can be used as a diagnostic tool for patients suffering from schizophrenia, ADHD, and bipolar disorder. The method includes three steps: (1) auditory stimulation with specific sound stimuli, (2) simultaneous measurement of brainstem activity, and (3) automated interpretation of the resulting brain stem audiograms with data-based signal analysis. We will compare three groups of 12 individuals with confirmed diagnoses of schizophrenia, ADHD, or bipolar disorder with 12 healthy subjects under blinded conditions for a total of 48 participants. The extent to which the method can be used to reach the correct diagnosis will be investigated. The project is now in a recruiting phase. When all patients and controls have been recruited and the measurements have been performed, the data will be analyzed according to a previously arranged algorithm. We expect the recruiting phase and measurements to be completed in early 2015, the analyses to be performed in mid-2015, and the results of the study to be published in early 2016. If the results support previous findings, this will lend strength to the idea that brainstem audiometry can offer objective diagnostic support for patients suffering from schizophrenia, ADHD, and bipolar disorder. A positive result from the study could imply that brainstem audiometry could become an important supportive tool for clinicians in their efforts to diagnose patients with these disorders in a timely and accurate manner. ClinicalTrials.gov NCT01629355; https://clinicaltrials.gov/ct2/show/NCT01629355 (Archived by WebCite at http://www.webcitation.org/6VBfTwx5H).


Computer Administered Safety Planning for Individuals at Risk for Suicide: Development and Usability Testing.
Safety planning is a brief intervention that has become an accepted practice in many clinical settings to help prevent suicide. Even though it is quick compared to other approaches, it frequently requires 20 min or more to complete, which can impede adoption. A self-administered, Web-based safety planning application could potentially reduce clinician time, help promote standardization and quality, and provide enhanced ability to share the created plan. The aim of this study was to design, build, and test the usability of a Web-based, self-administered safety planning application. We employed a user-centered software design strategy led by a multidisciplinary team. The application was tested for usability with a target sample of suicidal patients. Detailed observations, structured usability ratings, and Think Aloud procedures were used. Suicidal ideation intensity and perceived ability to cope were assessed pre-post engagement with the Web application. A total of 30 participants were enrolled. Usability ratings were generally strong, and all patients successfully built a safety plan. However, the completeness of the safety plan varied. The mean number of steps completed was 5.5 (SD 0.9) out of 6, with 90% (27/30) of participants completing at least 5 steps and 67% (20/30) completing all 6 steps. Some safety planning steps were viewed as inapplicable to some individuals. Some confusion in instructions led to modifications to improve understandability of each step. Ratings of suicide intensity after completion of the application were significantly lower than preratings, pre: mean 5.11 (SD 2.9) versus post: mean 4.46 (SD 3.0), t27=2.49, P=.02. Ratings of ability to cope with suicidal thoughts after completion of the application were higher than preratings, with the difference approaching statistical significance, pre: mean 5.93 (SD 2.9), post: mean 6.64 (SD 2.4), t27=-2.03, P=.05. We have taken the first step toward identifying the components needed to maximize usability of a self-administered, Web-based safety planning application. Results support initial consideration of the application as an adjunct to clinical contact. This allows for the clinician or other personnel to provide clarification, when needed, to help the patient build the plan, and to help review and revise the draft.


Defective platelet aggregation in myelodysplastic syndromes.
Hemorrhagic tendency in patients with myelodysplastic syndrome (MDS) is mainly attributed to thrombocytopenia. However, platelet function in these patients has not been thoroughly investigated. The aim of our study is to evaluate platelet function in patients with primary MDS. Platelet function was studied with aggregometry in response to ristocetin, collagen, ADP and adrenaline in 26 MDS patients and 15 healthy individuals. Aggregation was defective in 21 patients (80.7%). Adrenaline was the agonist with the most profound defect (45.9%), followed by ADP (58.7%), whereas aggregation with ristocetin and collagen was at the borderline. Abnormal aggregation to all four agonists was detected in 6 patients (23%). On the contrary, aggregation results were normal in only 5 patients (19.2%). RAEB-t (refractory anemia with excess blasts in transformation) patients were most seriously affected. MDS patients have impaired platelet aggregation in response to one or more stimulants. Platelet aggregation was not statistically different between MDS patients at early stages of the disease (<12 months) and those at later stages (>12 months). Defective platelet aggregation is strongly related to MDS of worse prognosis. None of our patients was detected to have hyperfunctional platelets, defined as platelets aggregating spontaneously. Functional defects in MDS do not elicit hemorrhagic tendency.


Expression of cytoplasmic islet cell antigens by rat pancreas.
A major problem in standardization of the islet cell cytoplasmic antibody (ICA) assay is variation in sensitivity of the different human pancreas substrates used in individual laboratories. To circumvent this problem, we have developed an assay that utilizes Wistar-Furth rat pancreas as substrate, an anti-islet monoclonal antibody (A2B5) to identify islets and fluorescein-conjugated protein A to identify patient autoantibodies. Sera from 85 control subjects, 27 type I diabetics, and 17 subjects at high risk for developing type I diabetes were assayed in parallel with our standard ICA assay on human pancreas substrate and with Wistar-Furth rat pancreas as substrate. Two sera from control subjects (2 of 85) were ICA positive with rat pancreas compared to 1 of 85 with human pancreas substrate. Sera from 11 of 27 type I diabetics and 15 of 17 sera from high-risk subjects were ICA positive with either rat or human pancreas substrate. A correlation between the specific islet fluorescence readings on human and rat pancreas sections was found with sera from high-risk and control subjects. Furthermore, end-point titers of an ICA-positive serum were identical with both assays. Finally, incubation of an ICA-positive serum with glycolipids, extracted from either human or Wistar-Furth rat pancreas, blocked subsequent ICA binding. These findings suggest that Wistar-Furth rat pancreas expresses an identical or similar autoantigen to human pancreas.


Interleukin-8, aquaporin-1, and inducible nitric oxide synthase in smoke and burn injured sheep treated with percutaneous carbon dioxide removal.
We previously showed that a percutaneous arteriovenous gas exchanger was effective in removing CO2 and reversing respiratory failure in an ovine model of adult respiratory distress syndrome (ARDS) produced by smoke inhalation and burn injury (Alpard et al., Ann Surg 230:215-224, 1999). In this study, we tested the hypothesis that arteriovenous CO2 removal (AVCO2R) lessened endogenous inflammation in the lung. Myeloperoxidase activity, aquaporin-1 (AQP-1), interleukin-8 (IL-8), and inducible nitric oxide synthase mRNAs as well as aquaporin-1, and IL-8 protein were measured in ovine lung tissue. Lung tissue was taken at 96 h (time of sacrifice) from animals with combined smoke inhalation and 40% third degree dermal burn and subsequently treated with AVCO2R or sham (ventilator alone) after onset of ARDS (PaO2:FiO2 ratio of < 200). Myeloperoxidase activity was 1.862 +/- 0.302 U/mg protein in the ventilator group and 0.830 +/- 0.141 in the AVCO2R plus ventilator group. AQP-1 mRNA was 140,482 +/- 31,702 copies/microg total RNA in the ventilator group and 61,854 +/- 22,433 copies/microg total RNA in the AVCO2R plus ventilator group (p = 0.076). mRNA for IL-8 mRNA in the ventilator alone treated animals was 74,000 +/- 3,300 copies/microg total RNA compared to < 1,000 copies/microg total RNA in the ventilator plus AVCO2R group. This result was highly significant (p < 0.001) Inducible nitric oxide synthase mRNA was 7,853 +/- 2,229 copies/microg total RNA for the AVCO2R group and 5,854 +/- 2,070 copies/microg total RNA for the ventilator managed animals. These differences were not statistically significant (p = 0.54). Percutaneous AVCO2R produced a specific decrease in IL-8 in the smoke and burn injured animals. Furthermore, this effect was consistent with cell signaling mechanisms that increase the expression of IL-8 by cyclic stretching and the observed reduction in the number of neutrophils in the lung parenchyma. Therefore, we speculate that the mechanism by which CO2 removal exerts a beneficial effect may be due to both decreases in ventilatory requirements, with an accompanying reduction in alveolar stretching, and reduction of neutrophil numbers in lung tissue.


Lithocholic bile acid accumulated in yeast mitochondria orchestrates a development of an anti-aging cellular pattern by causing age-related changes in cellular proteome.
We have previously revealed that exogenously added lithocholic bile acid (LCA) extends the chronological lifespan of the yeast Saccharomyces cerevisiae, accumulates in mitochondria and alters mitochondrial membrane lipidome. Here, we use quantitative mass spectrometry to show that LCA alters the age-related dynamics of changes in levels of many mitochondrial proteins, as well as numerous proteins in cellular locations outside of mitochondria. These proteins belong to 2 regulons, each modulated by a different mitochondrial dysfunction; we call them a partial mitochondrial dysfunction regulon and an oxidative stress regulon. We found that proteins constituting these regulons (1) can be divided into several "clusters", each of which denotes a distinct type of partial mitochondrial dysfunction that elicits a different signaling pathway mediated by a discrete set of transcription factors; (2) exhibit 3 different patterns of the age-related dynamics of changes in their cellular levels; and (3) are encoded by genes whose expression is regulated by the transcription factors Rtg1p/Rtg2p/Rtg3p, Sfp1p, Aft1p, Yap1p, Msn2p/Msn4p, Skn7p and Hog1p, each of which is essential for longevity extension by LCA. Our findings suggest that LCA-driven changes in mitochondrial lipidome alter mitochondrial proteome and functionality, thereby enabling mitochondria to operate as signaling organelles that orchestrate an establishment of an anti-aging transcriptional program for many longevity-defining nuclear genes. Based on these findings, we propose a model for how such LCA-driven changes early and late in life of chronologically aging yeast cause a stepwise development of an anti-aging cellular pattern and its maintenance throughout lifespan.


Two distinct auditory-motor circuits for monitoring speech production as revealed by content-specific suppression of auditory cortex.
Speech production, both overt and covert, down-regulates the activation of auditory cortex. This is thought to be due to forward prediction of the sensory consequences of speech, contributing to a feedback control mechanism for speech production. Critically, however, these regulatory effects should be specific to speech content to enable accurate speech monitoring. To determine the extent to which such forward prediction is content-specific, we recorded the brain's neuromagnetic responses to heard multisyllabic pseudowords during covert rehearsal in working memory, contrasted with a control task. The cortical auditory processing of target syllables was significantly suppressed during rehearsal compared with control, but only when they matched the rehearsed items. This critical specificity to speech content enables accurate speech monitoring by forward prediction, as proposed by current models of speech production. The one-to-one phonological motor-to-auditory mappings also appear to serve the maintenance of information in phonological working memory. Further findings of right-hemispheric suppression in the case of whole-item matches and left-hemispheric enhancement for last-syllable mismatches suggest that speech production is monitored by 2 auditory-motor circuits operating on different timescales: Finer grain in the left versus coarser grain in the right hemisphere. Taken together, our findings provide hemisphere-specific evidence of the interface between inner and heard speech.


Telecytology: Clinical applications, current challenges, and future benefits.
Telecytology is the interpretation of cytology material at a distance using digital images. For more than a decade, pioneering efforts to introduce telecytology into clinical practice have been reported. A Medline search for "telecytology" and "cytology" reveals a voluminous literature, though much of what has been published to date is based on technologies that are rapidly becoming obsolete. The technological limitations of previous techniques, including the transmission of static digital images and dynamic streaming images, have limited telecytology to minor niches. The primary problem with these technologies is that the remote viewer can only see a small fraction of the material on the original slides, introducing the possibility of diagnostic error based not only on image quality but also on image selection. Remote robotic microscopy offers one possible solution to this problem, but to date has found limited acceptance, principally attributable to slow operating times. Whole slide imaging seems to be a much more promising solution, though cytology-specific literature regarding its use is still scant. The advent of whole slide imaging opens up new possibilities for telecytology by enabling high-quality images of entire cytology specimens to be available to anyone, anywhere via the Internet. Although challenges remain, especially with regard to capturing the full microscopy experience including multiple planes of focus and sharp high-powered images, rapidly advancing technology promises to overcome these limitations. Increasing application of whole slide imaging technology in surgical pathology will undoubtedly also increase its application to cytology due to the increasing affordability and practicality of the equipment as it serves a larger number of useful roles within a pathology department. The current and expanding applications of telecytology for clinical practice, education, quality assurance, and testing will be reviewed.


Probiotic attributes and prevention of LPS-induced pro-inflammatory stress in RAW264.7 macrophages and human intestinal epithelial cell line (Caco-2) by newly isolated Weissella cibaria strains.
Probiotic lactic acid bacteria are known to modulate gut associated immune responses. Not many studies have reported on the role of Weissella species in preventing lipopolysaccharide (LPS) induced proinflammatory stress in murine macrophages as well as in human intestinal epithelial cells (Caco-2). Therefore, the present study was taken up to evaluate the probiotic attributes of four newly isolated Weissella strains (two each from fermented dosa batter and a human infant faecal sample); these attributes are cholesterol reduction, adhesion to Caco-2 cells and mucin and their ability to prevent LPS-induced nitric oxide and proinflammatory cytokine (IL-6, IL-1β and TNFα) production by the murine macrophages and IL-8 production by the human epithelial cells. Reduction in LPS induced pro-inflammatory stress was compared with a well-studied probiotic bacterium Lactobacillus rhamnosus GG. The results suggested that the strains were tolerant to gastric conditions (pH 3.0) and bile salts. In addition, the strains exhibited moderate cell surface hydrophobicity, cholesterol reduction and adhesion to Caco-2 cells and gastric mucin. All the strains could prevent LPS-induced nitric oxide and IL-6 production in murine macrophages, while strain 28 alone prevented IL-1β production. All the strains could prevent IL-8 production by the human epithelial cells. The present study led to the first line selection of W. cibaria 28 as a putative strain for future studies as it showed adhesion to Caco-2 cells and gastric mucin and cholesterol reduction besides preventing LPS-induced pro-inflammatory stress in macrophages and in human colonic epithelial cells.


Association of Testosterone Levels With Anemia in Older Men: A Controlled Clinical Trial.
In one-third of older men with anemia, no recognized cause can be found. To determine if testosterone treatment of men 65 years or older with unequivocally low testosterone levels and unexplained anemia would increase their hemoglobin concentration. A double-blinded, placebo-controlled trial with treatment allocation by minimization using 788 men 65 years or older who have average testosterone levels of less than 275 ng/dL. Of 788 participants, 126 were anemic (hemoglobin ≤12.7 g/dL), 62 of whom had no known cause. The trial was conducted in 12 academic medical centers in the United States from June 2010 to June 2014. Testosterone gel, the dose adjusted to maintain the testosterone levels normal for young men, or placebo gel for 12 months. The percent of men with unexplained anemia whose hemoglobin levels increased by 1.0 g/dL or more in response to testosterone compared with placebo. The statistical analysis was intent-to-treat by a logistic mixed effects model adjusted for balancing factors. The men had a mean age of 74.8 years and body mass index (BMI) (calculated as weight in kilograms divided by height in meters squared) of 30.7; 84.9% were white. Testosterone treatment resulted in a greater percentage of men with unexplained anemia whose month 12 hemoglobin levels had increased by 1.0 g/dL or more over baseline (54%) than did placebo (15%) (adjusted OR, 31.5; 95% CI, 3.7-277.8; P = .002) and a greater percentage of men who at month 12 were no longer anemic (58.3%) compared with placebo (22.2%) (adjusted OR, 17.0; 95% CI, 2.8-104.0; P = .002). Testosterone treatment also resulted in a greater percentage of men with anemia of known cause whose month 12 hemoglobin levels had increased by 1.0 g/dL or more (52%) than did placebo (19%) (adjusted OR, 8.2; 95% CI, 2.1-31.9; P = .003). Testosterone treatment resulted in a hemoglobin concentration of more than 17.5 g/dL in 6 men who had not been anemic at baseline. Among older men with low testosterone levels, testosterone treatment significantly increased the hemoglobin levels of those with unexplained anemia as well as those with anemia from known causes. These increases may be of clinical value, as suggested by the magnitude of the changes and the correction of anemia in most men, but the overall health benefits remain to be established. Measurement of testosterone levels might be considered in men 65 years or older who have unexplained anemia and symptoms of low testosterone levels. clinicaltrials.gov Identifier: NCT00799617.


Prevalence, evaluation and management of overactive bladder in primary care.
Patients with overactive bladder (OAB) are under-diagnosed in the primary care setting. Primary care physicians (PCP) approach to the patient and appropriate patient disclosure may contribute to under-diagnosis. An outpatient primary care setting was used to determine the prevalence and characteristics of OAB. Patients who visited the family medicine outpatient clinic were invited to answer a self-administered questionnaire. It included questions on evidence of lower urinary tract symptoms (modified Overactive Bladder-Validated 8-question Screener [OAB-V8]), relevant medical and surgical history, and demographic data. Relationship between OAB and other independent variables were analyzed using chi-square and risk ratio (RR) analysis. Of 325 questionnaires distributed, 311 were returned completed. Patients ranged from 18 to 97 years, the majority women (74.0%) and African American (74.3%). OAB was present in 60.5% of men and 48.3% of women (p = 0.058). OAB was significantly associated with obesity (BMI > or = 30) in women (p = 0.018, RR = 1.72), specifically obese premenopausal women (age < 55 years) (p = 0.011, RR = 1.98). OAB prevalence is more than double and higher in men than previously reported. The relative risk for OAB is significantly greater in obese premenopausal women.


Accessing the genomic information of unculturable oceanic picoeukaryotes by combining multiple single cells.
Pico-sized eukaryotes play key roles in the functioning of marine ecosystems, but we still have a limited knowledge on their ecology and evolution. The MAST-4 lineage is of particular interest, since it is widespread in surface oceans, presents ecotypic differentiation and has defied culturing efforts so far. Single cell genomics (SCG) are promising tools to retrieve genomic information from these uncultured organisms. However, SCG are based on whole genome amplification, which normally introduces amplification biases that limit the amount of genomic data retrieved from a single cell. Here, we increase the recovery of genomic information from two MAST-4 lineages by co-assembling short reads from multiple Single Amplified Genomes (SAGs) belonging to evolutionary closely related cells. We found that complementary genomic information is retrieved from different SAGs, generating co-assembly that features >74% of genome recovery, against about 20% when assembled individually. Even though this approach is not aimed at generating high-quality draft genomes, it allows accessing to the genomic information of microbes that would otherwise remain unreachable. Since most of the picoeukaryotes still remain uncultured, our work serves as a proof-of-concept that can be applied to other taxa in order to extract genomic data and address new ecological and evolutionary questions.


RNA degradation paths in a 12-subunit nuclear exosome complex.
The eukaryotic exosome is a conserved RNA-degrading complex that functions in RNA surveillance, turnover and processing. How the same machinery can either completely degrade or precisely trim RNA substrates has long remained unexplained. Here we report the crystal structures of a yeast nuclear exosome containing the 9-subunit core, the 3'-5' RNases Rrp44 and Rrp6, and the obligate Rrp6-binding partner Rrp47 in complex with different RNAs. The combined structural and biochemical data of this 12-subunit complex reveal how a single-stranded RNA can reach the Rrp44 or Rrp6 active sites directly or can bind Rrp6 and be threaded via the central channel towards the distal RNase Rrp44. When a bulky RNA is stalled at the entrance of the channel, Rrp6-Rrp47 swings open. The results suggest how the same molecular machine can coordinate processive degradation and partial trimming in an RNA-dependent manner by a concerted swinging mechanism of the two RNase subunits.


Home mechanical ventilation: outcomes according to remoteness from health center and different family education levels.
Throughout the world, home mechanical ventilation (HMV) is being increasingly employed to treat patients suffering from chronic respiratory failure. This present study aimed to examine the characteristics and outcomes of 27 children seen in our department over a four-year period who were treated with HMV. The causes of chronic respiratory failure were as follows: 16 (59.3%) neuromuscular disease, 6 (22.2%) primary respiratory diseases, 3 (11.1%) congenital heart disease, and 2 (7.4%) storage disease. The mean age was 59.4 months (1 day-15 years); mean follow-up for invasive ventilation was 356 (0-1200) days and for non-invasive HMV was 517 (30-1440) days. With respect to maternal educational level, 13 had graduated from elementary school and 14 from high school or university. Nine of our patients resided in Ankara, while 18 lived in rural areas of Turkey. Eleven of the 27 patients died during the HMV period (1-36 months) at home. Five patients were weaned from HMV between 1-19 months. Our experience showed that HMV can be applied successfully in chronic respiratory failure patients in Turkey. Length of the follow-up period and mortality rate were not affected by the patient's place of residence (city center or rural) or maternal level of education.


DNA condensation in live E. coli provides evidence for transertion.
Condensation studies of chromosomal DNA in E. coli with a tetranuclear ruthenium complex are carried out and images obtained with wide-field fluorescence microscopy. Remarkably different condensate morphologies resulted, depending upon the treatment protocol. The occurrence of condensed nucleoid spirals in live bacteria provides evidence for the transertion hypothesis.


Screening of predicted secreted antigens from Mycobacterium bovis reveals the immunodominance of the ESAT-6 protein family.
Results of previous studies utilizing bioinformatic approaches in antigen-mining experiments revealed that secreted proteins are among the most frequently recognized antigens from Mycobacterium bovis. Thus, we hypothesized that the analysis of secreted proteins is likely to reveal additional immunogenic antigens that can be used to increase the specificity of diagnostic tests or be suitable vaccination candidates for mycobacterial infections. To test this hypothesis, 382 pools of overlapping peptides spanning 119 M. bovis secreted and potentially secreted proteins were screened for the ability to stimulate a gamma interferon response in vitro using whole blood from tuberculin-positive reactor (TB reactor) cattle. Of the 119 proteins screened, 70 (59%) induced positive responses in the TB reactor animals to various degrees. Strikingly, all but one of the 15 ESAT-6 proteins tested were recognized by at least 30% of the TB reactor animals, with 12 of the 22 most commonly recognized antigens belonging to this protein family. Further analysis of these data demonstrated that there was no significant difference in immunogenicity between the ESAT-6 proteins that were components of potentially intact ESX secretory systems and those corresponding to additional partial esx loci. Importantly for vaccine design, antigenic epitopes in some highly conserved regions shared by numerous ESAT-6 proteins were identified. However, despite this considerable homology, peptide-mapping experiments also revealed that immunodominant peptides were located in regions of amino acid variability.


An fMRI study of reward-related probability learning.
The human striatum has been implicated in processing reward-related information. More recently, activity in the striatum, particularly the caudate nucleus, has been observed when a contingency between behavior and reward exists, suggesting a role for the caudate in reinforcement-based learning. Using a gambling paradigm, in which affective feedback (reward and punishment) followed simple, random guesses on a trial by trial basis, we sought to investigate the role of the caudate nucleus as reward-related learning progressed. Participants were instructed to make a guess regarding the value of a presented card (if the value of the card was higher or lower than 5). They were told that five different cues would be presented prior to making a guess, and that each cue indicated the probability that the card would be high or low. The goal was to learn the contingencies and maximize the reward attained. Accuracy, as measured by participant's choices, improved throughout the experiment for cues that strongly predicted reward, while no change was observed for unpredictable cues. Event-related fMRI revealed that activity in the caudate nucleus was more robust during the early phases of learning, irrespective of contingencies, suggesting involvement of this region during the initial stages of trial and error learning. Further, the reward feedback signal in the caudate nucleus for well-learned cues decreased as learning progressed, suggesting an evolving adaptation of reward feedback expectancy as a behavior-outcome contingency becomes more predictable.


First Report of Southern Blight of Ruellia brittoniana Caused by Sclerotium rolfsii in Louisiana.
Ruellia brittoniana, Mexican petunia, is an herbaceous flowering perennial grown in hardiness zones 8 to 10 in the southern and western United States. Popular dwarf forms with flower colors of white, pink, and blue are used as ground covers and borders. In April of 2003, root and stem rot that caused plant death was observed on cv. Katie (dwarf form, pink flowers) at a wholesale nursery in southern Louisiana. Plants were growing in a vermiculite and sand mix. The grower had purchased the plants from an out-of-state source, and approximately one-half of 1,440 plants were dead or dying. Symptoms included wilt, basal stem rot, and root rot. Peripheral roots were covered with a white mycelial layer that contained white sclerotial initials and small, brown sclerotia. Fungal isolates from infected roots grown on potato dextrose agar (PDA) produced white mycelia and 1- to 2-mm-diameter dark brown sclerotia. Sclerotia were nearly round with smooth surfaces and distributed over the entire colony. Isolates were identified as Sclerotium rolfsii on the basis of mycelial characteristics and color, size, and distribution of sclerotia. Two-month-old seedlings (6 to 10 cm high) of R. brittoniana, from seed of cv. Katie, were used in pathogenicity tests. Inoculum was grown in 10-cm-diameter plastic, culture dishes on PDA medium. Blended inoculum was prepared from a single 1-week-old culture that was composed of mycelia and sclerotia and blended 4 to 6 s at high speed in 100 ml of distilled water. In test one, 5 ml of inoculum was placed at the base of each inoculated plant. In test two, a single 5-mm-diameter agar plug with mycelium plus four sclerotia was placed beside plant stems near soil line. In test three, 5 ml of blended inoculum was dripped on exposed roots after plants were removed from pots. In test four, exposed plant roots were dipped in the blended inoculum. Each test contained 10 inoculated plants, and 10 noninoculated plants served as controls. All plants were placed in a dew chamber maintained at 28°C for 2 days and then returned to a greenhouse to observe development of symptoms and signs of disease. In tests one and two, basal stem rot and wilt developed on inoculated plants after 2 days and after 5 to 8 days all were dead. Inoculated plants from tests three and four were alive 4 months after inoculation, but were showing symptoms including leaf yellowing and drop, moderate to severe root rot, and some plants had begun to show white mycelia and white sclerotial initials on peripheral roots by January 2004. All noninoculated plants remained healthy and S. rolfsii was reisolated from infected plants in each test. To my knowledge, this is the first report of S. rolfsii causing disease on R. brittoniana.


Visible and near infrared fluorescence spectral flow cytometry.
There is a long standing interest in measuring complete emission spectra from individual cells in flow cytometry. We have developed flow cytometry instruments and analysis approaches to enable this to be done routinely and robustly. Our spectral flow cytometers use a holographic grating to disperse light from single cells onto a CCD for high speed, wavelength-resolved detection. Customized software allows the single cell spectral data to be displayed and analyzed to produce new spectra-derived parameters. We show that familiar reference and calibration beads can be employed to quantitatively assess instrument performance. We use microspheres stained with six different quantum dots to compare a virtual bandpass filter approach with classic least squares (CLS) spectral unmixing, and then use antibody capture beads and CLS unmixing to demonstrate immunophenotyping of peripheral blood mononuclear cells using spectral flow cytometry. Finally, we characterize and evaluate several near infrared (NIR) emitting fluorophores for use in spectral flow cytometry. Spectral flow cytometry offers a number of attractive features for single cell analysis, including a simplified optical path, high spectral resolution, and streamlined approaches to quantitative multiparameter measurements. The availability of robust instrumentation, software, and analysis approaches will facilitate the development of spectral flow cytometry applications.


Steroid psychosis caused by perioperative steroid replacement for pituitary adenoma: a case report.
Steroid psychosis is a frequent complication of steroid treatment. Although perioperative steroid replacement therapy is generally administered in patients undergoing pituitary surgery, there are no previous reports concerning the development of steroid psychosis after perioperative steroid replacement therapy following pituitary surgery. We herein report a case of steroid psychosis induced by perioperative steroid replacement therapy for pituitary surgery. A 35-year-old man presented with a visual disturbance that had persisted for 1 year. A magnetic resonance imaging scan showed a large pituitary tumor, and a laboratory study revealed slight dysfunction of the hypothalamo-pituitary-adrenal axis. The patient was diagnosed with a non-functioning pituitary tumor and underwent tumor resection via the endoscopic endonasal approach. The initial dose of perioperative steroid replacement therapy was 200 mg of hydrocortisone administered immediately before the operation. The replacement dose was gradually tapered and discontinued over a 7-day period. On postoperative day 4, the patient exhibited an elated mood, grandiose delusions, anxiety, and agitation. We diagnosed these psychiatric symptoms as steroid psychosis induced by steroid replacement and we prescribed risperidone as a treatment. The symptoms gradually improved and did not recur. This case highlights the risk of steroid psychosis following treatment with perioperative steroid replacement therapy for pituitary adenoma and raises questions regarding the appropriateness of perioperative steroid replacement for pituitary adenoma.


Mental health lived experience academics in tertiary education: the views of nurse academics.
Australian national mental health strategy emphasises inclusion of people diagnosed with mental illness in all areas of mental health care, policy development and education of health professionals. However, the way this inclusion has translated to Australian universities is relatively unexplored. Explore views of nurse academics regarding service user involvement in nursing education programmes. Qualitative exploratory. Australian universities offering educational programmes in nursing at postgraduate and undergraduate levels. Thirty four participants from 27 Australian universities participated. Data were collected using semi-structured telephone interviews with academics involved in teaching and/or coordinating undergraduate and/or postgraduate mental health nursing contents. Data were analysed using content analysis based on four cognitive processes: comprehending, synthesising, theorising and re-contextualising data. Four major themes emerged: good idea? long way to go; conceptualising the service user academic role; strengths of lived experience led student learning; and barriers to implementation. Findings indicated strong support for including mental health service users in teaching nursing students. However, at most universities service user engagement was often an informal arrangement, lacking clear guidelines and limited by financial barriers and the positioning of mental health nursing within curricula.


Quantifying killing of orangutans and human-orangutan conflict in Kalimantan, Indonesia.
Human-orangutan conflict and hunting are thought to pose a serious threat to orangutan existence in Kalimantan, the Indonesian part of Borneo. No data existed prior to the present study to substantiate these threats. We investigated the rates, spatial distribution and causes of conflict and hunting through an interview-based survey in the orangutan's range in Kalimantan, Indonesia. Between April 2008 and September 2009, we interviewed 6983 respondents in 687 villages to obtain socio-economic information, assess knowledge of local wildlife in general and orangutan encounters specifically, and to query respondents about their knowledge on orangutan conflicts and killing, and relevant laws. This survey revealed estimated killing rates of between 750 and 1800 animals killed in the last year, and between 1950 and 3100 animals killed per year on average within the lifetime of the survey respondents. These killing rates are higher than previously thought and are high enough to pose a serious threat to the continued existence of orangutans in Kalimantan. Importantly, the study contributes to our understanding of the spatial variation in threats, and the underlying causes of those threats, which can be used to facilitate the development of targeted conservation management.


Categorization and Comparison of Accessibility Testing Methods for Software Development.
There are many methods for testing accessibility and universal design, ranging from checklists and guidelines to automated testing and finally to human testing with participants from different user groups. It is, however, not straight forward to determine how a testing method relates to impairments and barriers. In this work we have expanded the W3C cognitive barriers from one category to four categories in order to provide a better overview of cognitive barriers and testing methods. We also present an overview of multiple existing accessibility testing methods and what kind of barriers they cover. Finally, we present a recommendation on how to select a collection of accessibility testing methods in order to cover the broadest range of disabilities. Our focus is tools that can empower and ease work processes of software development teams, including both developers and testers.


T regulatory cells and the control of alloimmunity: from characterisation to clinical application.
T regulatory cells (Treg) play an important role in the induction and maintenance of immunological tolerance. Recent findings in experimental transplant models combined with the development of functional reporter mice have opened new avenues to study Treg biology and their therapeutic potential. In particular, recent advances in understanding Treg function and lineage stability revealed unexpected plasticity of this lineage. Nevertheless, pre-clinical and pilot clinical trials using Treg cells as cellular therapies have been initiated suggesting the safety and feasibility of such treatment.


Structural encoding of human and schematic faces: holistic and part-based processes.
The range of specificity and the response properties of the extrastriate face area were investigated by comparing the N170 event-related potential (ERP) component elicited by photographs of natural faces, realistically painted portraits, sketches of faces, schematic faces, and by nonface meaningful and meaningless visual stimuli. Results showed that the N170 distinguished between faces and nonface stimuli when the concept of a face was clearly rendered by the visual stimulus, but it did not distinguish among different face types: Even a schematic face made from simple line fragments triggered the N170. However, in a second experiment, inversion seemed to have a different effect on natural faces in which face components were available and on the pure gestalt-based schematic faces: The N170 amplitude was enhanced when natural faces were presented upside down but reduced when schematic faces were inverted. Inversion delayed the N170 peak latency for both natural and schematic faces. Together, these results suggest that early face processing in the human brain is subserved by a multiple-component neural system in which both whole-face configurations and face parts are processed. The relative involvement of the two perceptual processes is probably determined by whether the physiognomic value of the stimuli depends upon holistic configuration, or whether the individual components can be associated with faces even when presented outside the face context.


Contaminants in stream sediments from seven United States metropolitan areas: part II--sediment toxicity to the amphipod Hyalella azteca and the midge Chironomus dilutus.
Relationships between sediment toxicity and sediment chemistry were evaluated for 98 samples collected from seven metropolitan study areas across the United States. Sediment-toxicity tests were conducted with the amphipod Hyalella azteca (28 day exposures) and with the midge Chironomus dilutus (10 day exposures). Overall, 33 % of the samples were toxic to amphipods and 12 % of the samples were toxic to midge based on comparisons with reference conditions within each study area. Significant correlations were observed between toxicity end points and sediment concentrations of trace elements, polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), or organochlorine (OC) pesticides; however, these correlations were typically weak, and contaminant concentrations were usually below sediment-toxicity thresholds. Concentrations of the pyrethroid bifenthrin exceeded an estimated threshold of 0.49 ng/g (at 1 % total organic carbon) in 14 % of the samples. Of the samples that exceeded this bifenthrin toxicity threshold, 79 % were toxic to amphipods compared with 25 % toxicity for the samples below this threshold. Application of mean probable effect concentration quotients (PECQs) based on measures of groups of contaminants (trace elements, total PAHs, total PCBs, OC pesticides, and pyrethroid pesticides [bifenthrin in particular]) improved the correct classification of samples as toxic or not toxic to amphipods compared with measures of individual groups of contaminants.


International Consensus Statement for the Screening, Diagnosis, and Treatment of Adolescents with Concurrent Attention-Deficit/Hyperactivity Disorder and Substance Use Disorder.
Childhood attention-deficit/hyperactivity disorder (ADHD) is a risk factor for substance misuse and substance use disorder (SUD) in adolescence and (early) adulthood. ADHD and SUD also frequently co-occur in treatment-seeking adolescents, which complicates diagnosis and treatment and is associated with poor treatment outcomes. Research on the effect of treatment of childhood ADHD on the prevention of adolescent SUD is inconclusive, and studies on the diagnosis and treatment of adolescents with ADHD and SUD are scarce. Thus, the available evidence is generally not sufficient to justify robust treatment recommendations. The aim of the study was to obtain a consensus statement based on a combination of scientific data and clinical experience. A modified Delphi study to reach consensus based upon the combination of scientific data and clinical experience with a multidisciplinary group of 55 experts from 17 countries. The experts were asked to rate a set of statements on the effect of treatment of childhood ADHD on adolescent SUD and on the screening, diagnosis, and treatment of adolescents with comorbid ADHD and SUD. After 3 iterative rounds of rating and adapting 37 statements, consensus was reached on 36 of these statements representing 6 domains: general (n = 4), risk of developing SUD (n = 3), screening and diagnosis (n = 7), psychosocial treatment (n = 5), pharmacological treatment (n = 11), and complementary treatments (n = 7). Routine screening is recommended for ADHD in adolescent patients in substance abuse treatment and for SUD in adolescent patients with ADHD in mental healthcare settings. Long-acting stimulants are recommended as the first-line treatment of ADHD in adolescents with concurrent ADHD and SUD, and pharmacotherapy should preferably be embedded in psychosocial treatment. The only remaining no-consensus statement concerned the requirement of abstinence before starting pharmacological treatment in adolescents with ADHD and concurrent SUD. In contrast to the majority, some experts required full abstinence before starting any pharmacological treatment, some were against the use of stimulants in the treatment of these patients (independent of abstinence), while some were against the alternative use of bupropion. This international consensus statement can be used by clinicians and patients together in a shared decision-making process to select the best interventions and to reach optimal outcomes in adolescent patients with concurrent ADHD and SUD.


Allogeneic gene-modified tumor cells (RCC-26/IL-7/CD80) as a vaccine in patients with metastatic renal cell cancer: a clinical phase-I study.
Despite novel targeted agents, prognosis of metastatic renal cell cancer (RCC) remains poor, and experimental therapeutic strategies are warranted. Transfection of tumor cells with co-stimulatory molecules and/or cytokines is able to increase immunogenicity. Therefore, in our clinical study, 10 human leukocyte antigen (HLA)-A(*)0201(+) patients with histologically-confirmed progressive metastatic clear cell RCC were immunized repetitively over 22 weeks with 2.5-40 × 10(6) interleukin (IL)-7/CD80 cotransfected allogeneic HLA-A(*)0201(+) tumor cells (RCC26/IL-7/CD80). Endpoints of the study were feasibility, safety, immunological and clinical responses. Vaccination was feasible and safe. In all, 50% of the patients showed stable disease throughout the study; the median time to progression was 18 weeks. However, vaccination with allogeneic RCC26/IL-7/CD80 tumor cells was not able to induce TH1-polarized immune responses. A TH2 cytokine profile with increasing amounts of antigen-specific IL-10 secretion was observed in most of the responding patients. Interferon-γ secretion by patient lymphocytes upon antigen-specific and non-specific stimulation was substantially impaired, both before and during vaccination, as compared with healthy controls. This is possibly due to profound tumor-induced immunosuppression, which may prevent induction of antitumor immune responses by the gene-modified vaccine. Vaccination in minimal residual disease with concurrent depletion of regulatory cells might be one strategy to overcome this limitation.


Estimating the cost of referral and willingness to pay for referral to higher-level health facilities: a case series study from an integrated community case management programme in Uganda.
Integrated community case management (iCCM) relies on community health workers (CHWs) managing children with malaria, pneumonia, diarrhoea, and referring children when management is not possible. This study sought to establish the cost per sick child referred to seek care from a higher-level health facility by a CHW and to estimate caregivers' willingness to pay (WTP) for referral. Caregivers of 203 randomly selected children referred to higher-level health facilities by CHWs were interviewed in four Midwestern Uganda districts. Questionnaires and document reviews were used to capture direct, indirect and opportunity costs incurred by caregivers, CHWs and health facilities managing referred children. WTP for referral was assessed through the 'bidding game' approach followed by an open-ended question on maximum WTP. Descriptive analysis was conducted for factors associated with referral completion and WTP using logistic and linear regression methods, respectively. The cost per case referred to higher-level health facilities was computed from a societal perspective. Reasons for referral included having fever with a negative malaria test (46.8%), danger signs (29.6%) and drug shortage (37.4%). Among the referred, less than half completed referral (45.8%). Referral completion was 2.8 times higher among children with danger signs (p = 0.004) relative to those without danger signs, and 0.27 times lower among children who received pre-referral treatment (p < 0.001). The average cost per case referred was US$ 4.89 and US$7.35 per case completing referral. For each unit cost per case referred, caregiver out of pocket expenditure contributed 33.7%, caregivers' and CHWs' opportunity costs contributed 29.2% and 5.1% respectively and health facility costs contributed 39.6%. The mean (SD) out of pocket expenditure was US$1.65 (3.25). The mean WTP for referral was US$8.25 (14.70) and was positively associated with having received pre-referral treatment, completing referral and increasing caregiver education level. The mean WTP for referral was higher than the average out of pocket expenditure. This, along with suboptimal referral completion, points to barriers in access to higher-level facilities as the primary cause of low referral. Community mobilisation for uptake of referral is necessary if the policy of referring children to the nearest health facility is to be effective.


Failure of clonal deletion in neonatally thymectomized mice: tolerance is preserved through clonal anergy.
Self-tolerance is achieved in part through intrathymic deletion of self-reactive T cells. The necessity of the thymus for this process is suggested by the development of autoimmune diseases in neonatally thymectomized (neoTx) mice and by the failure of clonal deletion in nude mice. Indeed, the present study demonstrates that neonatal thymectomy on day 3 after birth results in the failure of clonal deletion of V beta 11+ T cells in BALB/c mice and V beta 5+ and V beta 6+ T cells in DBA/2 mice. However, these potentially autoreactive cells are nonfunctional as measured by proliferation and lymphokine production after stimulation with appropriate anti-V beta mAbs or stimulator cells. It appears that this induction of nonresponsiveness may have occurred extrathymically: the early neonatal thymus (presumably the source of the peripheral T cells observed in neoTx mice) also contains T cells with self-reactive receptors, but these cells are fully functional. Therefore, neonatal thymectomy aborts deletion of self-reactive T cells, but self-tolerance is maintained through functional inactivation of potentially self-reactive clones.


Are laboratories following published recommendations for lupus anticoagulant testing? An international evaluation of practices.
Laboratory tests for lupus anticoagulants (LA) are commonly performed to evaluate thrombosis or suspected phospholipid antibody syndromes. To determine current LA testing practices, and if they conform to published recommendations, two questionnaires were distributed to clinical laboratory members of the North American Specialized Coagulation Laboratory Association (NASCOLA) and the ECAT Foundation (ECAT). The first and second questionnaires were completed by 113 and 96 laboratories, respectively. Commonly performed LA tests included the dilute Russell's viper venom time, LA sensitive activated partial thromboplastin time and hexagonal phospholipid test. Although some laboratories did single LA tests if requested, the majority complied with published recommendations: to use platelet poor plasma for LA tests; to use two or more screening tests, representing different assay principles, and one assay having a low phospholipid concentration to exclude LA; to confirm LA phospholipid dependency by the method giving an abnormal LA screen; to document the inhibitor activity on pooled normal plasma; and not to use phospholipid antibodies to confirm LA. A minority (<35%) followed the recommendations to exclude factor deficiencies and factor inhibitors as the cause of an abnormal LA test. After participating, 32% of laboratories had changed practices and 20% indicated that they would be changing practices. While most laboratories generally follow published guidelines for LA testing, few follow recommendations to evaluate for other coagulation abnormalities. Questionnaires may be helpful quality initiatives to improve compliance with laboratory testing guidelines and recommendations.


Generalized Lagrangian Path Approach to Manifestly-Covariant Quantum Gravity Theory.
A trajectory-based representation for the quantum theory of the gravitational field is formulated. This is achieved in terms of a covariant Generalized Lagrangian-Path (GLP) approach which relies on a suitable statistical representation of Bohmian Lagrangian trajectories, referred to here as GLP-representation. The result is established in the framework of the manifestly-covariant quantum gravity theory (CQG-theory) proposed recently and the related CQG-wave equation advancing in proper-time the quantum state associated with massive gravitons. Generally non-stationary analytical solutions for the CQG-wave equation with non-vanishing cosmological constant are determined in such a framework, which exhibit Gaussian-like probability densities that are non-dispersive in proper-time. As a remarkable outcome of the theory achieved by implementing these analytical solutions, the existence of an emergent gravity phenomenon is proven to hold. Accordingly, it is shown that a mean-field background space-time metric tensor can be expressed in terms of a suitable statistical average of stochastic fluctuations of the quantum gravitational field whose quantum-wave dynamics is described by GLP trajectories.


Human-level performance in 3D multiplayer games with population-based reinforcement learning.
Reinforcement learning (RL) has shown great success in increasingly complex single-agent environments and two-player turn-based games. However, the real world contains multiple agents, each learning and acting independently to cooperate and compete with other agents. We used a tournament-style evaluation to demonstrate that an agent can achieve human-level performance in a three-dimensional multiplayer first-person video game, Quake III Arena in Capture the Flag mode, using only pixels and game points scored as input. We used a two-tier optimization process in which a population of independent RL agents are trained concurrently from thousands of parallel matches on randomly generated environments. Each agent learns its own internal reward signal and rich representation of the world. These results indicate the great potential of multiagent reinforcement learning for artificial intelligence research.


Competency-based residency training: the next advance in graduate medical education.
The goal of all graduate medical education is to ensure that the graduating physician is competent to practice in his or her chosen field of medicine. The evaluation of a resident's competency to practice, however, has never been clearly defined, nor has the fixed period of time given for residency training in each specialty been shown to be the right amount of time for each individual resident to achieve competency. To better ensure that new physicians have the competencies they need, the author proposes the replacement of the current approach to residents' education, which specifies a fixed number of years in training, with competency-based training, in which each resident remains in training until he or she has been shown to have the required knowledge and skills and can apply them independently. Such programs, in addition to tailoring the training time to each individual, would make it possible to devise and test schemes to evaluate competency more surely than is now possible. The author reviews the basis of traditional residency training and the problems with the current training approach, both its fixed amount of time for training and the uncertainty of the methods of evaluation used. He then explains competency-based residency education, notes that it is possible, indeed probable, that some trainees will become competent considerably sooner than they would in the current required years of training, quotes a study in which this was the case, and explains the implications. He describes the encouraging experience of his neurosurgery department, which has used competency-based training for its residents since 1994. He then discusses issues of demonstrating competency in procedural and nonprocedural fields, as well as the evaluation of competency in traditional and competency-based training, emphasizing that the latter approach offers hope for better ways of assessing competency.


Associations between sedentary behavior and motor coordination in children.
This study was conducted to evaluate the relationship between objectively measured sedentary behavior (SB) and motor coordination (MC) in Portuguese children, accounting for physical activity (PA), accelerometer wear time, waist-to-height ratio, and mother's education level. A cross-sectional school-based study was conducted on 213 children (110 girls and 103 boys) aged 9-10 in the north of Portugal during the spring of 2010. Accelerometers were used to obtain detailed objective information about daily PA and SB over five consecutive days. MC was measured with a body coordination test (Körperkoordination Test für Kinder). Waist and height were measured by standardized protocols and the waist-to-height ratio (WHtR) was calculated. A questionnaire was used to assess mothers' educational levels. Receiver-operating characteristic (ROC) and logistic regressions were used. ROC analysis showed that sedentary time significantly discriminated between children with low MC and high MC, with a best trade off between sensitivity and specificity being achieved at ≥77.29% and ≥76.48% for girls and boys, respectively (P < 0.05 for both). In both genders, the low sedentary group had significantly higher odds of having good MC than the higher sedentary group, independent of PA, accelerometer wear time, WHtR, and mother's education level (P < 0.05 for both). Our findings suggested that PA levels per se may not overcome the deleterious influence of high levels of SB on MC. Our data stress the importance of discouraging SB among children to improve MC.


A binocular approach to treating amblyopia: antisuppression therapy.
We developed a binocular treatment for amblyopia based on antisuppression therapy. A novel procedure is outlined for measuring the extent to which the fixing eye suppresses the fellow amblyopic eye. We hypothesize that suppression renders a structurally binocular system, functionally monocular. We demonstrate using three strabismic amblyopes that information can be combined normally between their eyes under viewing conditions where suppression is reduced. Also, we show that prolonged periods of viewing (under the artificial conditions of stimuli of different contrast in each eye) during which information from the two eyes is combined leads to a strengthening of binocular vision in such cases and eventual combination of binocular information under natural viewing conditions (stimuli of the same contrast in each eye). Concomitant improvement in monocular acuity of the amblyopic eye occurs with this reduction in suppression and strengthening of binocular fusion. Furthermore, in each of the three cases, stereoscopic function is established. This provides the basis for a new treatment of amblyopia, one that is purely binocular and aimed at reducing suppression as a first step.


NCBO Resource Index: Ontology-Based Search and Mining of Biomedical Resources.
The volume of publicly available data in biomedicine is constantly increasing. However, these data are stored in different formats and on different platforms. Integrating these data will enable us to facilitate the pace of medical discoveries by providing scientists with a unified view of this diverse information. Under the auspices of the National Center for Biomedical Ontology (NCBO), we have developed the Resource Index-a growing, large-scale ontology-based index of more than twenty heterogeneous biomedical resources. The resources come from a variety of repositories maintained by organizations from around the world. We use a set of over 200 publicly available ontologies contributed by researchers in various domains to annotate the elements in these resources. We use the semantics that the ontologies encode, such as different properties of classes, the class hierarchies, and the mappings between ontologies, in order to improve the search experience for the Resource Index user. Our user interface enables scientists to search the multiple resources quickly and efficiently using domain terms, without even being aware that there is semantics "under the hood."


Enantiospecific adsorption of chiral molecules on chiral gold clusters.
Enantioselectivity in gold clusters is investigated by studying the adsorption of a chiral amino acid (cysteine) on a chiral Au55 cluster using density functional calculations. The highest adsorption energies were found when the amino and thiolate functional groups of cysteine are bonded to the lowest coordinated edges of the chiral cluster. Enantiospecific adsorption is primarily obtained from the different bond location and strength, at the cluster edge, of the carboxyl groups forming the left- and right-handed enantiomers. These results provide theoretical support to convey enantioselectivity in asymmetric nanocatalysts using chiral gold clusters.


Controlled, double-blind, randomized clinical trial to evaluate the impact of fruit juice consumption on the evolution of infants with acute diarrhea.
In order to assess the effects of juice feedings during acute diarrhea a double-blind, randomized study was performed in 90 children, mean age of 10 +/- 4.28 months. Thirty patients with acute diarrhea were fed twice-daily 15 ml/kg of Apple Juice (AJ), 30 received White Grape Juice (WGJ), and 30 were given colored and flavored water (WA) as part of their age appropriate dietary intake. The duration and severity of diarrhea were the main endpoint variables of the study performed in a metabolic unit. The patients were similar among the 3 groups, had diarrhea for 50-64 hours prior to admission, and were dehydrated when admitted to the unit for study. Half of the patients in each group were well nourished and the others had mild to moderate degrees of malnutrition. Rotavirus infection was the agent causing the illness in 63% of the patients. The infants fed juice ingested 14-17% more calories than those given WA, (those receiving AJ and WGJ ingested 95 and 98 Calories/Kg/d respectively) whereas those receiving WA consumed 81 cal/kg/d). The increased energy intake was not at the expense of other foods or milk formula. The mean body weight gain was greater among patients receiving WGJ (+ 50.7 gm) as compared with the patients in the AJ group (+ 18.3 gm) or the patients fed WA (- 0.7 gm) (p = 0.08). The duration of the illness was longer in the infants fed juice as compared with those given WA (p = 0.006), the mean +/- SD duration in hours was 49.4 +/- 32.6, 47.5 +/- 38.9 and 26.5 +/- 27.4 in patients fed AJ, WGJ and WA respectively. All patients improved while ingesting juice and none of them developed persistent diarrhea; most recovered within 50 hours of the beginning of treatment and less than one fourth had diarrhea longer than 96 hours in the unit. The fecal losses were also increased among the juice fed patients (p = 0.001); the mean +/- SD fecal excretion in g/kg/h was 3.94 +/- 2.35, 3.59 +/- 2.35, and 2.19 +/- 1.63 in AJ, WGJ and WA respectively. The stool output was highest during the first day of treatment among all the patients, though those fed AJ had the highest volume of fecal losses and those who received WA had the lowest stool excretion. After the first day of treatment the differences in fecal excretion were not significant. The ability to tolerate carbohydrates during the illness and immediately after recovery was similar among the 3 groups of patients. Intake of juices with different fructose/glucose ratios and osmolarities resulted in more fecal losses and more prolonged diarrhea as compared with water feedings, but the patients given juice ingested more calories and gained more weight, particularly among those being fed the juice with equimolar concentrations of fructose and glucose.