High diversity of small insectivorous mammals on Qinghai-Tibet Plateau and first description of karyotype for four endemics of China.
Among seven species of the order Eulipotyphla (from southern Gansu and northern Sichuan Provinces, Central China) studied cytogenetically, karyotypes of one talpid species, Uropsilus aff. soricipes (2n = 36, NFa = 54), and three soricid species, Chodsigoa hypsibia (2n = 65, NFa = 66), Sorex cansulus (2n = 42, NFa = 64) and Sorex thibetanus (2n = 42, NFa = 60), are described cytogenetically for the first time. All four species are endemic to China with distribution ranges restricted to the Qinghai-Tibet Plateau and adjacent mountain ranges. The Ch. hypsibia karyotype consists of mostly acrocentric autosomes and one metacentric pair of autosomes; besides, a B chromosome was identified. No polymorphism was detected among karyotypes of other species, including shrews Sorex bedfordiae (2n = 26, NFa = 44), Anourosorex squamipes (2n = 48, NFa = 92) and Crocidura suaveolens (2n = 40, NFa = 44). The Chinese shrew mole U. aff. soricipes and three shrew species (S. bedfordiae, Ch. hypsibia and A. squamipes) represent autochthonous fauna of Central/Western China, whereas S. thibetanus, S. cansulus and C. suaveolens belong to phylogenetic groups occurring mostly to the north and west from China; therefore, they should be considered relatively recent colonisers. Thus, considering the relationships of the species within phylogenetic groups, our results on karyotypes are in good agreement with molecular genetic data.


[Studies on coffee and coffee substitutes. XIX. Dependence of the quantity of a highly polymeric galactomannan on the degree of extraction coffee-extracts (author's transl)].
In extracts of Columbia Arabica Coffee the content of galactomannan precipitable with alcaline copper solution decreases slowly up to an extraction yield of 43.6%; it decreases somewhat more rapidly with higher extraction yields. An extract of Angola Robusta Coffee showed opposite behaviour. In this case the content of galactomannan increased with the extraction yield, although not evenly. Calculated on the basis of roasted coffee solids the extract of the Arabica coffee had a rather limited range precipitable polysaccharide, the Robusta coffee in contrast with rated increased amounts of galactomannan. The different behaviour of the two coffee varieties may be due to mote or less strong binding of this high-polymer carbohydrate to the cell wall.


a controlled trial of intratumoral ONYX-015, a selectively-replicating adenovirus, in combination with cisplatin and 5-fluorouracil in patients with recurrent head and neck cancer.
ONYX-015 is an adenovirus with the E1B 55-kDa gene deleted, engineered to selectively replicate in and lyse p53-deficient cancer cells while sparing normal cells. Although ONYX-015 and chemotherapy have demonstrated anti-tumoral activity in patients with recurrent head and neck cancer, disease recurs rapidly with either therapy alone. We undertook a phase II trial of a combination of intratumoral ONYX-015 injection with cisplatin and 5-fluorouracil in patients with recurrent squamous cell cancer of the head and neck. There were substantial objective responses, including a high proportion of complete responses. By 6 months, none of the responding tumors had progressed, whereas all non-injected tumors treated with chemotherapy alone had progressed. The toxic effects that occurred were acceptable. Tumor biopsies obtained after treatment showed tumor-selective viral replication and necrosis induction.


Identification by immunofluorescence of prolactin- and somatotrophin-producing cells in the pituitary gland of the tree frog Hyla arborea.
The indirect immunofluorescence procedure was used to localize prolactin (PRL)- and somatotrophin (STH)- producing cells in the pituitary distal lobe from Hyla arborea adult specimens. The following mammalian antisera were employed: rabbit anti-ovine PRL, antibovine PRL, anti-human PRL, anti-rat PRL, anti-ovine STH, anti-bovine STH, anti-human STH; monkey anti-rat STH. Immunocytochemical staining was suppressed by solid phase absorption of both anti-PRL and anti-STH with the specific antigen. Absorption of anti-PRL with STH and of anti-STH with PRL did not appreciably affect immunocytochemical staining. Treatment with the two antisera revealed two different reactive cell types, both acidophils. Using PRL antisera a strong fluorescence was found in the large acidophils located chiefly in the rostro-central and ventral areas of the distal lobe sagittal sections. A somewhat weaker fluorescence was observed using STH antisera in the fewer, small acidophils mostly concentrated in the dorso-caudal region and only sparsely scattered in the other areas of the pars distalis. Strikingly, the overall pattern of localization shown by the two cell types is similar to their already known distribution in the pituitary distal lobe of some other species of urodele and anuran amphibians.


Influence of season and anaesthetic on cutaneous vascular permeability responses to histamine, serotonin and mast cell-releasing agents in rats.
The influences of season and anaesthetic on the cutaneous vascular permeability responses to histamine (2.5 X 10(-8) mol), serotonin (5HT, 2.5 X 10(-10) mol), and to the mast cell-releasing agents adenosine 5(1)-triphosphate (2.5 X 10(-7) mol), dextran (5 X 10(-5) g) and compound 48/80 (1 X 10(-7) g) were examined using an Evans blue dye leakage technique. In rats anaesthetized with urethane (1 g/kg i.p.), responses to irritants were significantly less than in rats given ether for the period of administration of irritants and allowed to recover consciousness. The response to histamine was mediated by H1-receptors since it was abolished by mepyramine but not by metiamide. The only seasonal variation observed was in responses to 5HT in urethane-anaesthetized rats, which were significantly less in winter than in summer. It was concluded that dye leakage responses to irritants are affected more by anaesthetic than by seasonal variation.


An investigation into the concurrent collection of human scent and epithelial skin cells using a non-contact sampling device.
In criminal investigations, the collection of human scent often employs a non-contact, dynamic airflow device, known as the Scent Transfer Unit 100 (STU-100), to transfer volatile organic compounds (VOCs) from an object/person onto a collection material that is subsequently presented to human scent discriminating canines. Human scent is theorized to be linked to epithelial skin cells that are shed at a relatively constant rate allowing both scent and cellular material to be deposited into the environment and/or onto objects. Simultaneous collection of cellular material, with adequate levels of nuclear deoxyribonucleic acid (nDNA), and human scent using a non-invasive methodology would facilitate criminal investigations. This study evaluated the STU-100 for the concurrent collection of human scent and epithelial skin cells from a porous (paper) and non-porous (stainless steel bar) object that was held for a specified period of time in the dominant hand of twenty subjects (10 females and 10 males). Human scent analysis was performed using headspace static solid-phase microextraction with gas chromatography-mass spectrometry (HS-SPME/GC-MS). A polycarbonate filter was used to trap epithelial skin cells which, upon extraction, were subsequently analyzed, inter-laboratory, using the quantitative polymerase chain reaction (qPCR). The STU-100 proved to be inadequate for collecting the minimum number of epithelial skin cells required to obtain nuclear DNA concentrations above the limit of detection for the qPCR kit. With regard to its use for human scent collection, a reduction in the number and mass of compounds was observed when compared to samples that were directly collected. However, when the indirect collection of human scent from the two different objects was compared, a greater number and mass of compounds was observed from the non-porous object than from the porous object. This outcome suggests that the matrix composition of the scent source could affect the efficacy of the human scent collected when using a non-contact, dynamic airflow sampling device. The findings from this study are of importance because although the STU-100 proved to not be suitable for collecting epithelial skin cells for DNA analysis, its non-contact capability allows for the possibility of other potential forensic evidence, like that of human scent, to be obtained.


Effect of cryothermic and radiofrequency Cox-Maze IV ablation on atrial size and function assessed by 2D and 3D echocardiography, a randomized trial. To freeze or to burn.
Atrial linear scars in Cox-Maze IV procedures are achieved using Cryothermy (Cryo) or radiofrequency (RF) techniques. The subsequent postoperative left atrial (LA) reverse remodelling is unclear. We used 2- and 3-dimensional echocardiography (2-3DE) to compare the impact of Cryo and RF procedures on LA size and function 1 year after Cox-maze IV ablation concomitant with Mitral valve (MV) surgery. Seventy-two patients with MV disease and AF were randomized to Cryo (n = 35) or RF (n = 37) ablation. Another 33 patients were enroled without ablation (NoMaze). All patients underwent an echocardiogram the day before and 1 year after surgery. The LA function was assessed on 2D strain by speckle tracking and 3DE. Forty-two ablated patients recovered sinus rhythm (SR) 1 year after surgery. They had comparable left and right systolic ventricular function, LA volume index (LAVI), and 2D reservoir strain before surgery. At follow-up, the 3DE extracted reservoir and booster function were higher after RF (37 ± 10% vs. 26 ± 6%; p < 0.001) than Cryo ablation (18 ± 9 vs. 7 ± 4%; p < 0.001), while passive conduit function was comparable between groups (24 ± 11 vs. 20 ± 8%; p = 0.17). The extent of LAVI reduction depended on the duration of AF preoperatively. SR restoration after MV surgery and maze results in LA size reduction irrespective of the energy source used. Compared to RF, the extension of ablation area produced by Cryo implies a structural LA remodelling affecting LA systolic function.


Fetomaternal haemorrhage after external cephalic version at term.
The incidence of fetomaternal haemorrhage after external cephalic version was 1.8% in 167 patients. The occurrence of this complication was not found to be associated with difficult or unsuccessful version, or with any adverse perinatal outcome. We conclude that routine assessment of fetomaternal haemorrhage after external version is not necessary, except in rhesus negative women to detect the 2% in whom the routine dose of 500 iu (100 micrograms) of anti-D immunoglobulin is inadequate.


Pilonidal Disease: To Flap or Not to Flap.
Pilonidal disease is an acquired disease caused by infected hair follicles imbedded in the midline gluteal area. The diagnosis is made based on clinic examination. Treatment is based on disease severity with minimal invasive techniques like the Bascom I procedure, Moshe Gibs procedure, or video-assisted pilonidal sinus ablation best suited for less complicated cases. However, for recalcitrant diseases, more advanced techniques are needed such as the Karydakis, Limberg, or V-Y advancement flaps.


An oxygen-sensing mechanism for angiosperm adaptation to altitude.
Flowering plants (angiosperms) can grow at extreme altitudes, and have been observed growing as high as 6,400 metres above sea level1,2; however, the molecular mechanisms that enable plant adaptation specifically to altitude are unknown. One distinguishing feature of increasing altitude is a reduction in the partial pressure of oxygen (pO2). Here we investigated the relationship between altitude and oxygen sensing in relation to chlorophyll biosynthesis-which requires molecular oxygen3-and hypoxia-related gene expression. We show that in etiolated seedlings of angiosperm species, steady-state levels of the phototoxic chlorophyll precursor protochlorophyllide are influenced by sensing of atmospheric oxygen concentration. In Arabidopsis thaliana, this is mediated by the PLANT CYSTEINE OXIDASE (PCO) N-degron pathway substrates GROUP VII ETHYLENE RESPONSE FACTOR transcription factors (ERFVIIs). ERFVIIs positively regulate expression of FLUORESCENT IN BLUE LIGHT (FLU), which represses the first committed step of chlorophyll biosynthesis, forming an inactivation complex with tetrapyrrole synthesis enzymes that are negatively regulated by ERFVIIs, thereby suppressing protochlorophyllide. In natural populations representing diverse angiosperm clades, we find oxygen-dependent altitudinal clines for steady-state levels of protochlorophyllide, expression of inactivation complex components and hypoxia-related genes. Finally, A. thaliana accessions from contrasting altitudes display altitude-dependent ERFVII activity and accumulation. We thus identify a mechanism for genetic adaptation to absolute altitude through alteration of the sensitivity of the oxygen-sensing system.


Environment-friendly biomimetic synthesis of TiO₂ nanomaterials for photocatalytic application.
We have demonstrated an environment-friendly biomimetic synthesis method for the preparation of TiO(2) nanomaterials with different crystal phases and morphologies. This is the first time that it has been found that the crystal phase of TiO(2) can be controlled just by using different biotemplates, and cannot be changed by calcination up to 750 °C. In our experiment, anatase TiO(2) was obtained by using yeast and albumen templates, while rutile TiO(2) was formed by using dandelion pollen as the template.


The role of Hippo pathway signaling and A-kinase anchoring protein 13 in primordial follicle activation and inhibition.
To determine whether the mechanotransduction and pharmacomanipulation of A-kinase anchoring protein 13 (AKAP13) altered Hippo signaling pathway transcription and growth factors in granulosa cells. Primary ovarian insufficiency is the depletion or dysfunction of primordial ovarian follicles. In vitro activation of ovarian tissue in patients with primary ovarian insufficiency alters the Hippo and phosphatase and tensin homolog/phosphatidylinositol 3-kinase/protein kinase B/forkhead box O3 pathways. A-kinase anchoring protein 13 is found in granulosa cells and may regulate the Hippo pathway via F-actin polymerization resulting in altered nuclear yes-associated protein (YAP)/transcriptional coactivator with PDZ-binding motif coactivators and Tea domain family (TEAD) transcription factors. Laboratory studies. Translational science laboratory. None. COV434 cells, derived from a primary human granulosa tumor cell line, were studied under different cell density and well stiffness conditions. Cells were transfected with a TEAD-luciferase (TEAD-luc) reporter as well as expression constructs for AKAP13 or AKAP13 mutants and then treated with AKAP13 activators, inhibitors, and follicle-stimulating hormone. TEAD gene activation or inhibition was measured by TEAD-luciferase assays. The messenger ribonucleic acid levels of Hippo pathway signaling molecules, including connective tissue growth factor (CTGF), baculoviral inhibitors of apoptosis repeat-containing 5, Ankyrin repeat domain-containing protein 1, YAP1, and TEAD1, were measured by quantitative real-time polymerase chain reaction. Protein expressions for AKAP13, CTGF, YAP1, and TEAD1 were measured using Western blot. Increased TEAD-luciferase activity and expression of markers for cellular growth were associated with decreased cell density, increased well stiffness, and AKAP13 activator (A02) treatment. Additionally, decreased TEAD-luc activity and expression of markers for cellular growth were associated with AKAP13 inhibitor (A13) treatment, including a reduced expression of the BIRC5 and ANKRD1 (YAP-responsive genes) transcript levels and CTGF protein levels. There were no changes in TEAD-luc with follicle-stimulating hormone treatment, supporting Hippo pathway involvement in the gonadotropin-independent portion of folliculogenesis. These findings suggest that AKAP13 mediates Hippo-regulated changes in granulosa cell growth via mechanotransduction and pharmacomanipulation. The AKAP13 regulation of the Hippo pathway may represent a potential target for regulation of follicle activation.


Are xylem-tapping mistletoes partially heterotrophic?
Carbon isotope ratios, photosynthesis, and transpiration were measured on a xylem-tapping mistletoe (Phoradendron juniperinum) and its host (Juniperus osteosperma) in southern Utah, USA. For host tissues, the carbon isotope ratios agreed with theoretical values predicted from gas exchange observations. However, for mistletoe tissues, carbon isotope ratios deviated significantly from values predicted by gas exchange observations. This apparent discrepancy in mistletoe carbon isotope ratios can be resolved if one assumes that organic carbon dissolved in host xylem water was assimilated by the parasite. The mistletoes' high transpiration rates and low photosynthetic rates contributed to their heavy dependence on host xylem carbon. Two lines of evidence suggest that 62±2% of the carbon in the Utah mistletoe is derivated from the host and not from mistletoe autotrophic activities. Whereas xylem-tapping mistletoes have previously been characterized as wholly autotrophic parasites, we suggest that they may instead derive significant amounts of carbon from their hosts.


Detailed mapping of the phosphomannomutase 2 (PMM2) gene and mutation detection enable improved analysis for Scandinavian CDG type I families.
The gene for carbohydrate-deficient glycoprotein syndrome type I (CDG1) has previously been localised by us close to marker D16S406 in chromosome region 16p13.2-3. We also presented data indicating a strong founder mutation associated with a specific haplotype in CDG I patients from western Scandinavia. The phosphomannomutase 2 (PMM2) gene was recently put forward as a likely CDG1 candidate gene. We have now shown that the specific haplotype is associated with the PMM2 mutation 357C > A. Using data from radiation hybrid panel we have refined the position of the PMM2 gene to very close to marker D16S3020 in the interval between D16S406 and AFM282ze1 on the distal side and D16S3087 on the proximal side. Due to the severity of the disease many families request prenatal diagnostic services for CDG I. In the meantime, until the mutation spectrum is fully examined, we propose the combined use of mutation analysis and linkage analysis with polymorphic markers as diagnostic tools for Scandinavian CDG I families requesting prenatal diagnosis. Using this strategy we have to date successfully performed 15 prenatal diagnoses for CDG I.


The rs35874116 single nucleotide polymorphism increases sweet intake and the risk of severe early childhood caries: a case-control study.
Dental caries is one of the most prevalent chronic diseases worldwide among children. The rs35874116 single nucleotide polymorphism (SNP) in the sweet receptor gene TAS1R2 has been associated with dental caries at a high risk in permanent teeth among school children and adults. To date, little is known about the association of this SNP with sweet intake and caries risk in the primary school children. Total of 236 children were included, namely 118 subjects in the non-caries (NC) group and severe early childhood caries (S-ECC) group, respectively. Oral mucosa cells were collected from all the selected children, and the full length of exon 3 in TAS1R2 was sequenced to analyse rs35874116 polymorphism. A questionnaire was used to collect information about socio-demographic information, frequency of sweet intake and oral hygiene habits. Multiple logistic regression models were applied to assess the relationship of rs35874116 polymorphism with frequency of sweet intake and S-ECC among the five-year-old children. Children with the TT genotype of rs35874116 had a higher frequency of sweet intake than CT/CC carriers (51.3% vs. 32.7%; x2 = 5.436, p = 0.020), and S-ECC individuals were more likely to be TT genotype carriers than NC individuals (53.5% vs. 46.5%; x2 = 4.353, p = 0.037). Multiple logistic regression analyses showed that the TT genotype of rs35874116 was not only significantly related to the frequency of sweet intake (OR = 2.25, 95% CI = 1.14-4.44) but also significantly associated with S-ECC (OR = 2.11, 95% CI = 1.01 ~ 4.42). The rs35874116 polymorphism might increase sweet intake and the risk of S-ECC among five-year-old children in Nanning, China.


Effect of zinc ion on the interaction of some amino acid compounds of copper(II) with hydrogen peroxide.
Reaction of elemental copper and zinc powder mixtures with glycine (NH2.CH2COOH; HA) or aspartic acid (NH2CHCOOHCH2COOH; H2B) (in 1:1:2 ratio, respectively) in the presence of excess hydrogen peroxide (H2O2) at 50 degrees C, results in the formation of a new mixed metal peroxy carbonate compound corresponding to formula [Cu(Zn)2(O2(2-) (CO3)2(H2O)4], while the same reaction with elemental copper powder alone yields merely peroxy amino acid compounds having the formula [Cu(O2(2-)) (HA)2(H2O)] and [Cu(O2(2-)) (H2B) (H2O)2] for glycine and aspartic acid, respectively. These compounds have been characterized by elemental analysis, ESR, and electronic and IR spectra. It is interesting to note that both amino acids are converted to carbonate in the presence of zinc alone. A method analogous to that described above, for the reaction of elemental copper, zinc powder mixtures with succinic acid [(CH2COOH)2] or acetic acid (CH3COOH) in excess H2O2, on the other hand, gave a product essentially comprising copper succinate or acetate, respectively. These observations suggest an interesting and perhaps important phenomenon by which only the simple amino acids such as glycine and aspartic acid are converted to carbonates while their corresponding carboxylic acids form only their respective salts.


Phase-dependent locomotor activity in first-stadium nymphs of the desert locust, Schistocerca gregaria: effects of parental and progeny rearing density.
This study examined the effects of parental and progeny rearing density on locomotor activity of 1st-stadium nymphs of the desert locust, Schistocerca gregaria, using an actograph. Progeny obtained from solitarious (isolated-reared) or gregarious (crowd-reared) locusts were reared in isolation or in a group of 30 nymphs. Crowding after hatching had a slight influence on mean activity shortly after the start of measurements, but no clear effect was detected until day 2, when maximum activity during the 6-24 h of observation was significantly higher than that of the nymphs kept in isolation. On the other hand, the effects of parental rearing density on locomotor activity manifested at all ages examined (0-2 days old). Progeny of gregarious locusts showed consistently higher activity than those of solitarious locusts. In newly hatched nymphs, the effect of parental rearing density was explained by variation in body size at hatching, one of the phase-dependent characteristics. Hatchling body color was also correlated with locomotor activity and body weight. Similar levels of locomotor activity were exhibited when green, solitarious and black, gregarious nymphs were similar in body weight. These results suggested that parental rearing density indirectly influences locomotor activity in the progeny shortly after hatching by affecting their body size as eggs or hatchlings.


Aversiveness of errors and the error-related negativity (ERN): A systematic review on the affective states' manipulations findings.
Error-related negativity (ERN) has been used to investigate neural mechanisms underlying error processing and conflict monitoring. Recent evidence highlights that affective and motivational states modulate the ERN and that aversiveness of errors plays a vital role in error monitoring. Therefore, our primary objective was to systematically evaluate and describe the influence of affect state-related manipulations on the ERN. A total of 51 publications identified from PsyInfo, PubMed, and PsyArticles databases were included following the Prisma procedures for systematic reviews. Papers were analyzed using sample attributes, psychological paradigms, and states manipulations. The present study shows that the ERN component has recurrently appeared to be sensitive to manipulations of affective states in the reviewed literature. However, conclusive findings concerning the affect state-dependent properties of the ERN remain elusive. Results are discussed considering heterogeneity in paradigms, variables, and the state-trait interactions. Furthermore, recommendations for future high-quality studies are provided along with the necessity of upcoming high-power replication attempts and more studies with positive affect manipulations.


Roots of smile: a preterm neonates' study.
Twenty-two preterm neonates were observed 1h per neonate in the NICU. Ninety-five spontaneous smiles were recorded. Younger and smaller neonates showed more and longer spontaneous smiles than older and larger. The youngest neonate was 200 days from conception on the observational day. She was 511 g. This infant showed spontaneous smiles. The roots of spontaneous smiles are discussed.


Comparison of the 2010 and 2005 versions of the McDonald MRI criteria for dissemination-in-time in Taiwanese patients with classic multiple sclerosis.
In 2010, the International Panel on the Diagnosis of Multiple Sclerosis revised the 2005 version of the McDonald criteria. The revisions to MRI dissemination-in-time criteria include adoption of a new criterion by demonstration of simultaneous asymptomatic gadolinium-enhancing and nonenhancing lesions on baseline MRI scans. The purpose of this study was to demonstrate the diagnostic validity of the modified MRI dissemination-in-time criteria. We collected 80 patients with an initial clinical attack suggestive of an acute central nervous system demyelinating disease. The patients were followed for at least two years or until the development of definite multiple sclerosis. The nonconverters were taken as negative cases. Their baseline and follow-up brain MRI studies were retrospectively reviewed by two neuroradiologists. The 2010 version had higher sensitivity (68.2% vs. 45.5%), slightly lower specificity (80.6% vs. 83.3%), and higher accuracy (73.8% vs. 62.5%) than the 2005 version, but the differences were without statistical significance. The new criteria are more sensitive and accurate and specific just as the old criteria. They allow the diagnosis of definite multiple sclerosis in 34.1% patients at first presentation of the clinically isolated syndrome.


The transfer of analytical procedures.
Analytical method transfers are certainly among the most discussed topics in the GMP regulated sector. However, they are surprisingly little regulated in detail. General information is provided by USP, WHO, and ISPE in particular. Most recently, the EU emphasized the importance of analytical transfer by including it in their draft of the revised GMP Guideline. In this article, an overview and comparison of these guidelines is provided. The key to success for method transfers is the excellent communication between sending and receiving unit. In order to facilitate this communication, procedures, flow charts and checklists for responsibilities, success factors, transfer categories, the transfer plan and report, strategies in case of failed transfers, tables with acceptance limits are provided here, together with a comprehensive glossary. Potential pitfalls are described such that they can be avoided. In order to assure an efficient and sustainable transfer of analytical procedures, a practically relevant and scientifically sound evaluation with corresponding acceptance criteria is crucial. Various strategies and statistical tools such as significance tests, absolute acceptance criteria, and equivalence tests are thoroughly descibed and compared in detail giving examples. Significance tests should be avoided. The success criterion is not statistical significance, but rather analytical relevance. Depending on a risk assessment of the analytical procedure in question, statistical equivalence tests are recommended, because they include both, a practically relevant acceptance limit and a direct control of the statistical risks. However, for lower risk procedures, a simple comparison of the transfer performance parameters to absolute limits is also regarded as sufficient.


Engineering a thermostable version of D-allulose 3-epimerase from Rhodopirellula baltica via site-directed mutagenesis based on B-factors analysis.
D-allulose has received increasing attention due to its excellent physiological properties and commercial potential. The D-allulose 3-epimerase from Rhodopirellula baltica (RbDAEase) catalyzes the conversion of D-fructose to D-allulose. However, its poor thermostability has hampered its industrial application. Site-directed mutagenesis based on homologous structures in which the residuals on high flexible regions were substituted according to B-factors analysis, is an effective way to improve the thermostability and robustness of an enzyme. RbDAEase showed substrate specificity toward D-allulose with a Km of 58.57 mM and kcat of 1849.43 min-1. It showed a melting temperature (Tm) of 45.7 °C and half-life (t1/2) of 52.3 min at pH 8.0, 60 °C with 1 mM Mn2+. The Site-directed mutation L144 F strengthened the thermostability to a Δt1/2 of 50.4 min, ΔTm of 12.6 °C, and ΔT5060 of 22 °C. It also improved the conversion rate to 28.6%. Structural analysis reveals that a new hydrophobic interaction was formed by the mutation. Thus, site-directed mutagenesis based on B-factors analysis would be an efficient strategy to enhance the thermostability of designed ketose 3-epimerases.


Interaction of total N-nitroso compounds in environment and in vivo on risk of esophageal cancer in the coastal area, China.
Shantou, China, is one of the endemic regions of esophageal cancer (EC) in the coastal areas, but the relationship between the multiple exposures of total N-nitroso compounds (TNOCs) and the high risk of EC to date is unclear. This study is to estimate the potential integrative role of multiple exposures of environmental and endogenous TNOC on the EC risk. We collected randomly the samples of 72 h diet and 24 h urine from 256 healthy male (aged 30-65 years) in the high- and low-risk area for EC, and determined the contents of TNOC and 4 N-nitrosamino acids (NAAs) by a method of chemical fission-Thermal Energy Analyzer (TEA) and gas chromatography--TEA, respectively. We used TNOC and NAAs index to evaluate the integrative effect of multiple TNOC exposures on the EC risk by a logistic model. TNOC intake and urinary TNOC content were higher in the high-risk subjects than that in the low-risk subjects by 1.60 micromol/day and 1.45 micromol/L, respectively. TNOC intake, urinary TNOC content and its interaction were significant factors involved in the higher risk of EC (R(2)=0.685, p=0.000) with predicted Correct Class of 85.6%. The risk of the occurrence of a larger SMR of EC may increase by 4.841-fold for every 1 micromol/day increase in TNOC intake (95% CI 2.370-9.888, p=0.000) and by 4.113-fold for every 1 micromol/L increase in urinary TNOC content (95% CI 2.108-8.023, p=0.000), respectively. The integrally role of multiple exposures of TNOC in environment and in vivo may increase the risk of EC in the coastal areas in eastern Guangdong, China.


Solvent strength of aqueous phase for two typical biphasic solvent systems in high-speed countercurrent chromatography.
In the present work, influence of solvent strength of aqueous phase for two frequently-used biphasic solvent system in partition coefficient (K) of selected solutes were mainly studied, and a new method for selection of biphasic solvent system was proposed for high-speed countercurrent chromatographic separations. Solvent strength was referred to the typical theory that was deeply investigated in conventional reversed-phase liquid chromatography. Experimental results showed that a linear relationship between log(K) of solutes and apparent content of methanol in biphasic solvent system was found for the biphasic solvent system hexane-ethyl acetate-methanol-water (HEMWat), which was consistent with the relationship between real content and apparent content of methanol in this system. Meanwhile, a quadratic relationship was found between log(K) of solutes and apparent content of methanol in biphasic solvent system chloroform-methanol-water (ChMWat), in which it was found that the relationship between real content and apparent content of methanol in this system was also quadratic. In addition, a visual and simple method was proposed to select a suitable biphasic solvent system for separation of target compounds by high-speed countercurrent chromatography with isocratic elution, which saves a lot of manpower and material resources in order to find a suitable two-solvent system. An optimal biphasic solvent system for isolation of several tested compounds by high-speed countercurrent chromatography was easily obtained using our proposed method.


Senescent Microglia Represent a Subset of Disease-Associated Microglia in P301S Mice.
The existence and contribution of microglia with senescent-like alterations in the pathogenesis of age-related neurodegenerative diseases like Alzheimer's disease (AD) have been suggested in recent years. However, the identification of this distinct microglial population in vivo has proven challenging, largely due to overlaps in the inflammatory phenotype of activated and senescent microglia. Furthermore, attempts at recapitulating senescence in microglia in vitro are limited. To identify and characterize senescent microglia that occur in vivo in an animal model of neurodegeneration driven by pathologic tau. We analyzed the RNA expression patterns of individual microglia from normal mice and the pathogenic tau P301 S PS19 mouse model. We have previously demonstrated that p16-expressing senescent microglia occur in these mice when neurodegeneration has occurred. Here we identify a subset of disease-associated microglia with senescent features, notably characterized by the expression of Ccl4. This signature overlaps with established markers of senescence from other cell types. Our characterization of senescent microglia can be used to better understand the role of senescent microglia in various age-related contexts, including whether clearance of senescent microglia represents a viable therapeutic option.


Nuclear uptake control of NF-kappa B by MAD-3, an I kappa B protein present in the nucleus.
I kappa B proteins specifically inhibit the DNA binding of NF-kappa B/Rel transcription factors. An additional role as inhibitors of nuclear uptake was supposed by subcellular fractionation and enucleation experiments. Using indirect immunofluorescence labeling of cells, we show here that the DNA-binding p50 and p65 subunits of NF-kappa B, as well as the I kappa B protein MAD-3, all occur in the nucleus when overexpressed on their own. Nuclear uptake of p65 and, to a lesser extent of p50, was, however, suppressed when MAD-3 was coexpressed. Likewise, nuclear uptake of MAD-3 was blocked by overexpressed p65 or p50. This directly demonstrates that I kappa B is a nuclear uptake regulatory protein and that the various subunits of NF-kappa B can mutually control their access to the nucleus. In the presence of MAD-3, antibodies specific for peptides overlapping the nuclear location signal (NLS) sequences of p65 and p50 could not recognize their epitopes on NF-kappa B, suggesting that the I kappa B protein rendered the signals inaccessible for NLS receptors.


Tidal effects on periodical variations in the occurrence of singing humpback whales in coastal waters of Chichijima Island, Ogasawara, Japan.
Marine organisms inhabiting coastal waters are known to be driven by periodic cycles such as diel, tidal, and seasonal changes. Humpback whales (Megaptera novaeangliae) breed in shallow and warm coastal waters, with males singing complex songs during the breeding season. To investigate periodic variations in humpback whale singing activities, we conducted fixed passive acoustic monitoring in the Ogasawara (Bonin) Islands, Japan, from winter to spring during 2016-2018. The singing activity and individual number of singers were observed throughout the day and night using a very long baseline passive acoustic array. The occurrence of singers peaked before sunrise and in the evening and was reduced during the daytime. The frequency of song reception depended on the tidal phase. A generalised additive model demonstrated that the occurrence of singers increased during the flood tide and decreased during the ebb tide in the waters west of Chichijima Island. These results suggest that the singing behaviour of humpback whales in breeding areas is affected by the diel and tidal cycles. Male humpback whales may change their behaviour or singing location depending on the strength and direction of the tidal current, considering that the selection of a stable location is beneficial for singing whales.


An Intriguing Method for Fabricating Arbitrarily Shaped "Matreshka" Hydrogels Using a Self-Healing Template.
This work describes an intriguing strategy for the creation of arbitrarily shaped hydrogels utilizing a self-healing template (SHT). A SHT was loaded with a photo-crosslinkable monomer, PEG diacrylate (PEGDA), and then ultraviolet light (UV) crosslinked after first shaping. The SHT template was removed by simple washing with water, leaving behind the hydrogel in the desired physical shape. A hierarchical 3D structure such as "Matreshka" boxes were successfully prepared by simply repeating the "self-healing" and "photo-irradiation" processes. We have also explored the potential of the SHT system for the manipulation of cells.


Plasma biomarkers outperform echocardiographic measurements for cardiovascular risk prediction in kidney transplant recipients: results of the HOME ALONE study.
Since kidney transplant recipients (KTRs) have a high cardiovascular disease burden, adequate risk prediction is of importance. Whether echocardiographic parameters and plasma biomarkers, natriuretic peptides [N-terminal pro-B-type natriuretic peptide (NT-proBNP)] and troponin T provide complementary or overlapping prognostic information on cardiovascular events remains uncertain. The prospective Heterogeneity of Monocytes and Echocardiography Among Allograft Recipients in Nephrology (HOME ALONE) study followed 177 KTRs for 5.4 ± 1.7 years. Predefined endpoints were hospitalization for acute decompensated heart failure or all-cause death (HF/D) and major atherosclerotic cardiovascular events or all-cause death (MACE/D). At baseline, plasma NT-proBNP, plasma troponin T and echocardiographic parameters [left atrial volume index, left ventricular (LV) mass index, LV ejection fraction, and LV filling pressure] were assessed. Among all echocardiographic and plasma biomarkers measured, only NT-proBNP was consistently associated with HF/D in univariate and multivariate {third versus first tertile: hazard ratio [HR] 4.20 [95% confidence interval (CI) 1.02-17.27]} analysis, and only troponin T was consistently associated with MACE/D in univariate and multivariate [third versus first tertile: HR 8.15 (95% CI 2.75-24.18)] analysis. Our data suggest that plasma biomarkers are robust and independent predictors of heart failure and atherosclerotic cardiovascular events after kidney transplantation, whereas standard echocardiographic follow-up does not add to risk prediction.


Resolving the paradigm crisis in intravenous iron and erythropoietin management.
Despite the proven benefits of intravenous (i.v.) iron therapy in anemia management, it remains underutilized in the hemodialysis population. Although overall i.v. iron usage continues to increase slowly, monthly usage statistics compiled by the US Renal Data System suggest that clinicians are not implementing continued dosing regimens following repletion of iron stores. Continued therapy with i.v. iron represents a key opportunity to improve patient outcomes and increase the efficiency of anemia treatment. Regular administration of low doses of i.v. iron prevents the recurrence of iron deficiency, enhances response to recombinant human erythropoietin therapy, minimizes fluctuation of hemoglobin levels, hematocrit levels, and iron stores, and may reduce overall costs of care. This article reviews the importance of i.v. iron dosing on a regular basis in the hemodialysis patient with iron-deficiency anemia and explores reasons why some clinicians may still be reluctant to employ these protocols in the hemodialysis setting.


Effects of disinfectants on dimensional accuracy of impression materials.
There is a continuing concern over the potential for cross-contamination of viral infections such as AIDS by means of dental impressions. This investigation compared the effects of various viricidal agents on the accuracy of irreversible hydrocolloid and silicone impressions made of single-unit and full mandibular arch models. Measurements on casts made from the treated impressions were compared with the corresponding measurements on casts made from untreated impressions. The results of this study led to the conclusions that there was no preferred disinfectant with respect to dimensional stability for the irreversible hydrocolloid, condensation silicone, or vinyl polysiloxane impression materials tested, and there was no difference in the accuracy of the casts obtained either by spraying with or immersion in any of the disinfectants tested. Therefore using the disinfectants described with either the immersion or spray techniques will not significantly affect the dimensional accuracy of cats made from treated impressions.


Skeletally Immature Patients With Adolescent Idiopathic Scoliosis Curves 15°-24° Are at High Risk for Progression.
Retrospective review. To evaluate the incidence of adolescent idiopathic scoliosis (AIS) curve progression and brace prescription in skeletally immature patients (Risser 0 to Risser 1) with curves 15°-24°. Many skeletally immature patients with mild AIS ask about the likelihood of curve progression. No studies have answered these questions. The charts and radiographs of 302 consecutive patients with curves 15°-24° at initial visit, Risser 0 to Risser 1, were reviewed until skeletal maturity (≥Risser 4) or surgery. Curves averaged 19.1° ± 2.9° at initial visit. The Risser grade was 0 in 247 patients (82%) and 1 in 55 patients (18%). Patients who were Risser 0 were compared with those who were Risser 1, curves 15°-19° were compared with curves 20°-24°. The majority of patients demonstrated curve progression ≥5° (65%). Patients who were Risser 0 did not progress significantly more than patients who were Risser 1 (10° vs. 8°) (p = .22). Patients with curves 20°-24° did not progress significantly more than patients with curves 15°-19° (10° vs. 9°) (p = .65). Curve progression for small curves (15°-19°) is similar to curves between 20° and 24°. Close observation or perhaps early intervention for these patients is necessary. These data may suggest a paradigm shift to earlier brace initiation and call for early treatment in small curves. Level II.


Recovery of cognitive function after coronary artery bypass graft operations.
The effect of coronary artery bypass grafting (CABG) operations on cognition was examined after controlling for the operation, emotional state, preexisting cognitive impairment, and repeated experience with cognitive tests. On-pump CABG patients (n=16), thoracic surgical patients (n=15), and a nonsurgical control group (n=15) were tested preoperatively, and at 1 and 8 weeks postoperatively, using a battery of cognitive tests and an emotional state assessment. Patient groups were similar in age, sex, level of education, and premorbid intelligence quotient score. Surgical group data were normalized against data from the nonsurgical control group before statistical analysis. CABG patients performed worse on every subtest before the operation, and this disadvantage persisted after the operation. Anxiety, depression, and stress were associated with impaired cognitive performance in the surgical groups 1 week after the operation: 44% of CABG patients and 33% of surgical control patients were significantly impaired; yet, by 8 weeks, nearly all patients had recovered to preoperative levels, with 25% of CABG and 13% of surgical control patients improving beyond their preoperative performance. Stress, anxiety, and depression impair cognitive performance in association with CABG and thoracic operations. Most patients recover to, or exceed, preoperative levels of cognition within 8 weeks. Thus, after controlling for nonsurgical factors, the prospects of a tangible improvement in cognition after CABG are high.


Asymmetries of prefrontal cortex in human episodic memory: effects of transcranial magnetic stimulation on learning abstract patterns.
Functional neuroimaging suggests asymmetries of memory encoding and retrieval in the prefrontal lobes, but different hypotheses have been presented concerning the nature of prefrontal hemispheric specialization. We studied an associative memory task involving pairs of Kanji (Chinese) pictographs and unfamiliar abstract patterns. Subjects were ten Japanese adults fluent in Kanji, so only the abstract patterns represented novel material. During encoding, transcranial magnetic stimulation (TMS) was applied over the left and right dorsolateral prefrontal cortex (DLPFC). A significant (P<0.05) reduction in subsequent recall of new associations was seen only with TMS over the right DLPFC. This result suggests that the right DLPFC contributes to encoding of visual-object associations, and is consistent with a material-specific rather than a process-specific model of mnemonic function in DLPFC.


Association between 2 indexes for evaluating malocclusion in the mixed dentition.
This study aimed to associate 2 indexes for evaluating malocclusion in the mixed dentition. A population-based cross-sectional study was conducted involving 743 children aged 8-10 years, divided into early and late mixed dentition. The Dental Aesthetic Index (DAI) and criteria proposed by Grabowski were used to evaluate malocclusion. Associations between the DAI (outcome variable) and the other independent variables (gender, race, and mixed dentition stage) were analyzed using logistic regression models. Simple logistic regression models were constructed, estimating the crude odds ratios with the respective 95% confidence intervals (CIs). The variables with P < 0.20 in the simple analyses were tested in multiple logistic regression models, estimating adjusted odds ratios with the respective 95% CIs. We found that 84.01% (185/220) of the diagnoses were coincidental between the Grabowski Index and DAI in patients without malocclusion. In contrast, 59.80% (439/734) of the patients were diagnosed by both indexes (Grabowski and DAI) with malocclusion. Children with malocclusion, diagnosed by the Grabowski Index, were 5.85 (P = 0.0001 and 95% CI: 4.08-8.39) times more likely to show malocclusions when diagnosed by the DAI. DAI and the criteria proposed by Grabowski were expressed differently in relation to malocclusion in the mixed dentition. This study suggests the importance of specific criteria for the evaluation of malocclusion at this stage of development.


Assessment of neural inhibition of the lower esophageal sphincter in cats with esophagitis.
The aim of this study was to investigate the inhibitory innervation of the lower esophageal sphincter in the presence of esophagitis. Esophagitis was produced in five anesthetized cats with intraesophageal perfusion of HCl. Sphincter pressure responses were assessed with a sleeve catheter after administration of bethanechol, cholecystokinin octapeptide, and McNeil-A343 and with intraesophageal balloon distension. In the presence of esophagitis (1) resting lower esophageal sphincter pressure decreased; (2) the excitatory response to bethanechol was maintained; (3) there was a reduction in the excitatory response to McNeil-A343 and cholecystokinin at the highest dosages; (4) there was an increase in the potency of cholecystokinin and McNeil-A343 to produce an inhibitory response; (5) the inhibitory response to intraesophageal balloon distension was maintained; and (6) increased inhibitory responses took longer to normalize than the reduced excitatory responses. Esophagitis decreases cholinergic excitation, but neural inhibition to the LES remains intact. These findings suggest that blocking intact inhibition may be a new therapeutic approach for esophagitis caused by gastroesophageal reflux.


Computed tomography in evolution of testicular cancer during intensive chemotherapy.
The evolution of malignant testicular tumor to mature teratoma has been studied in 4 patients. Computed tomography has been helpful in early diagnosis of this biologic phenomenon in these patients receiving intensive chemotherapy for disseminated non-seminomatous testicular cancer. Although the potential significance of this conversion in terms of survival is not known its early recognition by computed tomography has been useful in selecting and monitoring the therapy of these patients.


Self-reported knee pain and disability among healthy individuals: reference data and factors associated with the Knee injury and Osteoarthritis Outcome Score (KOOS) and KOOS-Child.
To develop normative reference data for the Knee injury and Osteoarthritis Outcome Score (KOOS) and KOOS-Child, as well as investigate socio-demographic, psychological and physical factors associated with knee pain and disability among healthy adults. The KOOS or KOOS-Child (each containing five subscales) was administered to participants aged 8-101 years within the 1000 Norms Project, an observational study of 1000 self-reported healthy individuals. Self-efficacy, physical activity, body mass index (BMI), lower limb alignment, knee frontal plane projection angle (FPPA), knee range of motion (ROM), knee and hip strength, six-minute walk, 30-second chair stand and timed up and down stairs tests were collected. KOOS data were dichotomised using established cut-off scores and logistic regression analyses were conducted for each subscale. Socio-demographic characteristics were similar to the Australian population. Normative reference data were generated for children (8-17 years) and adults (18-101 years). Female adults were up to twice as likely to report knee pain, symptoms and sport/recreation (Sport/Rec) limitations compared to males (P < .05). Older age, lower self-efficacy, greater BMI, varus lower limb alignment, lower knee flexion ROM and lower hip external rotation (ER) strength were independently associated with knee pain and disability among adults. Age- and gender-stratified reference data for the KOOS and KOOS-Child have been developed to guide interpretation of results in practice and research for individuals with knee disorders. Psychological and physical factors are linked with self-reported knee pain/disability among adults, and longitudinal studies to investigate causation are required.


Singlet molecular oxygen generated in dark biological process.
Ultraweak chemiluminescence arising from biomolecules oxidation has been attributed to the radiative deactivation of singlet molecular oxygen [(1)O2] and electronically excited triplet carbonyl products involving dioxetane intermediates. As examples, we will discuss the generation of (1)O2 from lipid hydroperoxides, which involves a cyclic mechanism from a linear tetraoxide intermediate. The generation of (1)O2 in aqueous solution via energy transfer from the excited triplet acetone arising from the thermodecomposition of dioxetane a chemical source, and horseradish peroxidase-catalyzed oxidation of 2-methylpropanal, as an enzymatic source, will also be discussed. The approach used to unequivocally demonstrate the generation of (1)O2 in these reactions is the use of (18)O-labeled hydroperoxide / triplet dioxygen ((18)[(3)O2]), the detection of labeled compounds by HPLC coupled to tandem mass spectrometry (HPLC-MS/MS) and the direct spectroscopic detection and characterization of (1)O2 light emission. Characteristic light emission at 1,270nm, corresponding to the singlet delta state monomolecular decay was observed. Using(18)[(3)O2], we observed the formation of (18)O-labeled (1)O2 ((18)[(1)O2]) by the chemical trapping of (18)[(1)O2]with the anthracene-9,10-diyldiethane-2,1-diyl disulfate disodium salt (EAS) and detected the corresponding (18)O-labeled EAS endoperoxide usingHPLC-MS/MS. The combined use of the thermolysis of a water-soluble naphthalene endoperoxide as a generator of (18)O labeled (1)O2 and the sensitivity of HPLC-MS/MS allowed the study of (1)O2reactivity toward biomolecules. Photoemission properties and chemical trapping clearly demonstrate that the production of hydroperoxide and excited carbonyls generates (18)[(1)O2], and points to the involvement of (1)O2 in physiological and pathophysiological mechanism. Supported by FAPESP (2012/12663-1), CAPES, INCT Redoxoma (FAPESP/CNPq/CAPES; 573530/2008-4), NAP Redoxoma (PRPUSP; 2011.1.9352.1.8), CEPID Redoxoma (FAPESP; 2013/07937-8).


Role of mouse polyomavirus late region in the control of viral DNA replication: a review.
The genome of polyomaviruses is divided into two coding regions: the early and the late region. A relatively short regulatory sequence, encompassing the origin of viral DNA replication (ori), separates the two regions encoding the structural genes. In mouse polyomavirus (Py) in particular, the early DNA codes for three antigens: large, middle and small T-antigen (L-T, M-T and S-T, respectively). Large T antigen binds ori and thus regulates both viral DNA transcription and replication. Middle T antigen has been shown to mediate malignant transformation in non-permissive cells in vitro. No defined function has been assigned to the small T antigen although this gene product is thought to act synergistically both with L- and M-T antigens. The viral late region of Py encodes also three different genes whose products form the viral capsid during the productive infection cycle in permissive cells. Py early region was thought to be the only part of the genome necessary to code for proteins of functional and regulatory significance. The viral late region, on the other hand, was for a long time considered a simple reservoir of structural information, since it codes for capsid proteins and was supposedly devoid of functional control properties. This short review is focused on recent works from our and other laboratories, reporting evidence that in Py also the late region has a functional role since late sequences are involved in the control of viral DNA replication and in capsid assembly. Results indicating that this might be true for the cognate simian virus SV40 will be also reviewed.


Human immunodeficiency virus-1 infection and multiple sclerosis-like illness in a child.
We present a child with human immune deficiency virus-1 infection associated with a multiple sclerosis-like illness. This case is the first reported of human immune deficiency virus-1 infection and a disease indistinguishable from multiple sclerosis in a child. Because of the rarity of either disease in pediatric patients, their coexistence in a child argues in favor of a casual association between the two illnesses, thus reinforcing the long-held assumption that viral agents may trigger some of the immunologic abnormalities underlying multiple sclerosis. Cases in which such an association was present in adults are also described in this article.


Nanochitosan functionalized hydrophobic starch/guar gum biocomposite for edible coating application with improved optical, thermal, mechanical, and surface property.
The current work demonstrates a unique approach of utilizing nanochitosan (NCS) based edible nanomodifier for functionalizing starch (ST)/guar gum (GG) biocomposite with superior packaging properties targeting stringent edible food packaging on fresh cuts. The effectiveness of NCS in terms of structure-property-performance analysis of ST/GG biocomposites was done. The inclusion of NCS to the biocomposites of ST/GG converts its hydrophilic surface nature to hydrophobic (contact angle of ~114°) by modifying the surface features. The addition of NCS improved the thermal stability, where the observed 10% weight degradation of ST biocomposites were ~79.36, ~80.49, and ~186.89 °C for neat ST, ST/GG biocomposites, and ST/GG/NCS (3% w/v) (ST-GG-NCS3), respectively. The observed transparency of ST, ST-GG, and ST-GG-NCS3 were 21, 8, and 48%, respectively in the visible region suggesting consumer preference for transparent packaging materials. The wt% of O, C and N elements in ST-GG-NCS3 as observed by EDX spectra were ~ 50.2, ~47.6, and ~ 2.2%, respectively, which confirm the safety of the materials. Additionally, it is noteworthy to mention that the storage quality in terms of microbial growth, pH change, color attributes, and weight loss are better preserved when used as an edible coating on cut apple fruits.


Pathophysiologic Origins of Brachial Plexus Injury.
To analyze the origins and consequences of cases of brachial plexus injuries and their relationship to shoulder dystocia. We conducted a retrospective cohort study and identified all neonates with brachial plexus injury delivered at our institution between March 2012 and July 2019. A review was performed of the maternal and neonatal records of each neonate to identify obstetric antecedents, including the occurrence of shoulder dystocia and persistence of the injury. Experience of the delivering clinician was also examined. Statistical analysis was performed with the Fisher exact test, χ test for trends, and two-tailed t tests. Thirty-three cases of brachial plexus injury were identified in 41,525 deliveries (0.08%). Fourteen (42%) of these cases were not associated with shoulder dystocia; three (9%) followed cesarean delivery. Brachial plexus injury without shoulder dystocia was related to the absence of maternal diabetes, lower birth weights, and a longer second stage of labor. Persistent brachial plexus injury at the time of discharge was seen with equal frequency among neonates with (17/19, 89%, 95% CI 0.52-100%) and without shoulder dystocia (10/14, 71%, 95% CI 34-100%), P=.36). Whether brachial plexus injury was transient or persistent after shoulder dystocia was unrelated to the years of experience of the delivering clinician. Despite ongoing training and simulation, the already low incidence of brachial plexus injury did not decrease over time at our institution. Brachial plexus injury and shoulder dystocia represent two complications of uterine forces driving a fetus through the maternal pelvis in the presence of disproportion between the passage and the shoulder girdle of the passenger. Either or both of these complications may occur, but often are not causally related.


Effect of induced ventricular fibrillation and shock delivery on brain natriuretic peptide measured serially following a predischarge ICD test.
Brain natriuretic peptide (BNP) was a marker for heart failure and cardiac wall tension. We analysed the trend of BNP after predischarge testing in order to get non-invasive details about the cardiac stress during predischarge testing. 4-5 days after ICD implant we measured BNP, myoglobin, cardiac troponin I and creatine kinase in 20 patients before and 1, 5, 10, 20, 40, 60, 80, 100, 120 minutes and at the next day after predischarge testing. We evaluated actual values and percentage alterations of BNP. BNP significantly increased with a maximum after 5 minutes (804.0 +/- 803.4 vs. 475.7 +/- 629.5 pg/ml, P < 0.0001) and in terms of the percentage values (100 vs. 199.4 +/- 61.4 %, P < 0.0001) compared with baseline BNP. BNP decreased after that with the last significantly increased BNP value after 20 minutes (540.2 +/- 604.9 vs. 475.7 +/- 629.5 pg/ml, P = 0.017). We excluded a cardiac necrosis during predischarge testing because of similar values of myoglobin, cardiac troponin I and creatine kinase during the 2-hour follow-up. Our data showed a great increase with a doubling of BNP after 5 minutes as a result of induced ventricular fibrillation during predischarge test. This increase was not generated by myocardial necrosis but rather caused by an acute cardiac failure as a consequence of induced ventricular fibrillation in predischarge testing.


Age-related changes in cerebellar noradrenergic pre- and postsynaptic mechanisms: intrinsic vs extrinsic determinants evaluated with brain grafts in oculo.
Intrinsic versus extrinsic determinants of changes in cerebellar noradrenergic transmission during senescence in the rat were measured using homologous cerebellar grafts in oculo. Postsynaptic sensitivity of Purkinje neurons to catecholamines was determined by perfusing the anterior eye chamber with known concentrations of norepinephrine (NE) dissolved in a balanced salt solution. NE elicited a dose-dependent slowing of spontaneous Purkinje neuron discharge in both young (3-6 months) and aged (20-22 months) cerebellar grafts. Hill plots demonstrated that the dose-response relationships in both age-groups were linear and parallel to one another. Aged transplant Purkinje neurons manifested a marked and highly significant subsensitivity to NE with an EC50 of 583 microM, as compared with an EC50 of only 15.9 microM in the young grafts. Young grafts in 15-21-month-old hosts manifested an EC50 of 20 microM for the depressant actions of NE. Collaterals of host iris sympathetic fibers innervate the grafts. Activity of these fibers can be reflexly altered by changing illumination of the retina. The dynamics of presynaptic NE release from these fibers was evaluated using in vivo electrochemistry with Nafion-coated graphite epoxy capillary electrodes, which are highly selective for the monoamine neurotransmitters. As illumination of the ipsilateral retina is increased, the release of catecholamine in the cerebellar graft decreases. A mean change in the extracellular electroactive species of 4.2 +/- 0.6 microM was found in young cerebellar grafts. Equivalent stimuli induced a mean change of 2.3 +/- 0.8 microM in aged grafts. However, this diminished release was not statistically significant.(ABSTRACT TRUNCATED AT 250 WORDS)


Road safety evaluation with multiple treatments: A comparison of methods based on simulations.
This paper focuses on ex-post road safety evaluation with multiple treatments. The potential outcome framework for causal inference is introduced to formalize the causal estimands of interest. Various estimation methods are compared via performing simulation experiments based on semi-synthetic data constructed from a London 20 mph zones dataset. The methods under evaluation include regressions, propensity score (PS) based methods, and a machine learning-based method termed generalized random forests (GRF). Both PS-based methods and GRF show higher flexibility with respect to functional specifications of outcome models. Moreover, GRF shows great superiority in the cases where road safety treatments are assigned following specific criteria and/or where there are heterogeneous treatment effects. Considering the ex-post evaluation of combined effects of multiple treatments has significant practical value, the potential outcome framework and the estimation methods presented in this paper are highly recommended for road safety studies.


CSF monoamine metabolites and neuropeptides in depressed patients before and after electroconvulsive therapy.
Antidepressant drugs affect monoamines and neuropeptides in human cerebrospinal fluid (CSF) and in rodent brain. The purpose of this study was to investigate if also electroconvulsive therapy (ECT) affects these compounds in a similar manner in the CSF of depressed patients. Homovanillic acid (HVA), 5-hydroxyindoleacetic acid (5-HIAA), and corticotropin-releasing hormone (CRH)-like immunoreactivity (-LI) and neuropeptide Y (NPY)-LI were determined in CSF in six drug resistant patients with major depression. Lumbar puncture was performed at baseline and after completion of eight ECTs. ECT was associated with an increase in NPY-LI (p=0.009) and a decrease in CRH-LI (p<or=0.001). HVA (p=0.003) and 5-HIAA (p=0.002) were significantly increased after the ECT. Findings of NPY increase and CRH decrease were similar to those following chronic treatment with citalopram, indicating that these changes might constitute one of the common underpinnings of antidepressant treatment modalities.


Multiple side effects of Efalizumab in a Saudi female with chronic persistent psoriasis followed by severe rebound after Efalizumab discontinuation.
We are reporting a case of 23-year-old Saudi female with persistent chronic plaque psoriasis who was given subcutaneous Efalizumab 0.8 mg/kg/week for 14 weeks. During that period the patient developed multiple adverse reactions followed by severe rebound. The case is presented to highlight the importance of managing patients on Efalizumab carefully and closely.


Different Culture Conditions Affect Drug Transporter Gene Expression, Ultrastructure, and Permeability of Primary Human Nasal Epithelial Cells.
This study aimed to characterize a commercially available primary human nasal epithelial cell culture and its gene expression of a wide range of drug transporters under different culture conditions. Human nasal cells were cultured in three different types of culture media at the air-liquid (A-L) or liquid-liquid (L-L) interfaces for 1 or 3 wks. The effects of the different cell culture conditions were evaluated using light and electron microscopy, transepithelial electrical resistance (TEER) measurements, permeation studies with dextran, and gene expression profiling of 84 drug transporters. The type of culture medium affected cell ultrastructure, TEER, and dextran permeation across epithelia. The expression of 20 drug transporter genes depended on the culture interface and/or time in culture; the A-L interface and longer time in culture favored higher expression levels of five ABC and seven SLC transporters. Culture conditions influence the morphology, barrier formation, permeation properties, and drug transporter expression of human nasal epithelial cells, and this must be taken into consideration during the establishment and validation of in vitro models. A thorough characterization of a nasal epithelial model and its permeability properties is necessary to obtain an appropriate standardized model for the design of aerosol therapeutics and drug transport studies.


Omega-3 polyunsaturated fatty acids prevent atrial fibrillation associated with heart failure but not atrial tachycardia remodeling.
There is epidemiological evidence that omega-3 polyunsaturated fatty acids (PUFAs) reduce the risk of atrial fibrillation (AF), but clinical data are conflicting. The present study assessed the effects of PUFA on AF in experimental models. We studied the effects of oral PUFA supplements in 2 experimental AF paradigms: electrical remodeling induced by atrial tachypacing (400 bpm for 1 week) and congestive heart failure-associated structural remodeling induced by ventricular tachypacing (240 bpm for 2 weeks). PUFA pretreatment did not directly change atrial effective refractory period (128+/-6 [mean+/-SEM] versus 127+/-2 ms; all effective refractory periods at 300-ms cycle lengths) or burst pacing-induced AF duration (5+/-4 versus 34+/-18 seconds). Atrial tachypacing dogs had shorter refractory periods (73+/-6 ms) and greater AF duration (1185+/-300 seconds) than shams (119+/-5 ms and 20+/-11 seconds; P<0.01 for each). PUFAs did not significantly alter atrial tachypacing effects on refractory periods (77+/-8 ms) or AF duration (1128+/-412 seconds). PUFAs suppressed ventricular tachypacing-induced increases in AF duration (952+/-221 versus 318+/-249 seconds; P<0.05) and attenuated congestive heart failure-related atrial fibrosis (from 19.2+/-1.1% to 5.8+/-1.0%; P<0.001) and conduction abnormalities. PUFAs also attenuated ventricular tachypacing-induced hemodynamic dysfunction (eg, left ventricular end-diastolic and left atrial pressure from 12.2+/-0.5 and 11.4+/-0.6 mm Hg, respectively, to 6.4+/-0.5 and 7.0+/-0.8 mm Hg; P<0.01) and phosphorylation of mitogen-activated protein kinases (extracellular-signal related and P38 kinase). PUFAs suppress congestive heart failure-induced atrial structural remodeling and AF promotion but do not affect atrial tachycardia-induced electrical remodeling. The beneficial effects of PUFAs on structural remodeling, possibly related to prevention of mitogen-activated protein kinase activation, may contribute to their clinical anti-AF potential.


Discrete dinuclear complex to extended 2D compound in a Cu-azido system by controlling coligand stoichiometry: synthesis and magneto-structural correlations.
This article describes syntheses, structural characterizations and magnetic studies of two different Cu(II)-azido compounds, a discrete dinuclear complex and an extended 2D network. The compounds, [Cu(μ(1,1)-N3)(N3)(Me2en)]2 (1) and [Cu3(μ(1,1,1)-N3)2(μ(1,1,3)-N3)(μ(1,1)-N3)2(μ(1,3)-N3)(Me2en)]n (2), have been synthesized by controlling the relative concentration of the blocking ligand, N,N-dimethylethylenediamine (Me2en). Compound 1 is a dinuclear compound which is formed by a doubly asymmetric μ(1,1)-N3 bridging ligand, while 2 is a rare Cu-azido system where four different types of binding modes of azide ligands are present in a single compound. Compound 2 contains a hexanuclear core, where the Cu(II) centres are connected to each other by μ(1,1,1), μ(1,1) and μ(1,1,3) bridging azide ligands. The hexanuclear core acts as a secondary building block and further assembles via μ(1,3) and μ(1,1,3) azide groups, forming a 2D network in the crystallographic ac plane. Interestingly, temperature-dependent magnetic study suggests that the dinuclear compound 1 exhibits an antiferromagnetic interaction through the μ(1,1)-N3 bridge, which has also been supported by density functional theory (DFT) calculations. In the case of 2, an overall dominant ferromagnetic interaction is observed while antiferromagnetic interaction operates between the hexanuclear cores.


Purification and properties of a dicyclohexylcarbodiimide-sensitive adenosine triphosphatase from a thermophilic bacterium.
1. A stable ATPase complex with sensitivity to dicyclohexylcarbodiimide (TFo-F1) was purified from the membranes of the thermophilic aerobic bacterium PS3, by ion exchange chromatography in the presence of Triton X-100. 2. The ATPase of TFo-F1 was maximal at 70 degrees at pH 8.6 and was stable after monomerization in 4 M urea and 0.5% Triton X-100 at 25 degrees. The activity was dependent on Mg2+, Co2+, or Mn2+, and it became insensitive to dicyclohexylcarbodiimide when Ca2+ or Cd2+ was added instead. 3. TFo-F1 required P-lipids of this bacterium contained branched fatty acyl groups but no unsaturated groups and were stable against oxidation and heat. 4. Studies by electron microscopy, gel electrophoresis, and use of anti-ATPase antibody and [3H]acetyl-ATPase indicated that the TFo-F1 complex was composed of an ATPase moiety (TF1, five different subunits) and a hydrophobic moiety (TFo, three different subunits. TFo conferred TF1 with sensitivity to dicyclohexylcarbodiimide. 5. Vesicles catalyzing 32Pi-ATP exchange and ATP-driven enhancement of fluorescence of anilinonaphthalene sulfonate were reconstituted by dialyzing pure TFo-F1 and P-lipids together, and were active even at 50-75 degrees. The vesicles reconstituted from TFo-F1 and bacterial P-lipids were more stable than those reconstituted from TFo-F1 and soybean P-lipids.


Predictor of enhanced mortality in patients with multimorbidity and atrial fibrillation in an acute hospital setting.
Patients with atrial fibrillation (AF) admitted to hospital commonly have comorbidities. Few studies have attempted to determine factors prognostic of mortality in hospitalized AF patients with multimorbidity. To identify factors associated with mortality in hospitalized AF patients. Retrospective cohort study. Patients with multimorbidity (≥2 chronic diseases), with or without AF, discharged from Lugo hospital (Spain) between 1 January 2000 and 31 December 2015. Data were extracted from hospital medical records. Of 74 220 patients (170 978 hospitalizations), 52 939 had multimorbidity (14 181 had AF; 38 758 no AF) and were included in our study. Patients with AF were older (mean ± standard deviation 78.6 ± 10.0 vs. 71.9 ± 14.2 years) and had a higher mortality rate (27.1 vs. 20.5%) than those without AF. Gender (female), age, stroke and congestive heart failure (CHF), but not AF, were independently associated with mortality. AF significantly increased the mortality risk in women [relative risk (RR) 1.091; 95% confidence interval (CI) 1.021-1.165; P = 0.010] and in those aged >80 years (RR 1.153; 95% CI, 1.1-1.2; P < 0.001). CHF independently increased the risk of mortality across all age groups (RR 1.496; 95% CI 1.422-1.574; P < 0.001). Hospitalized patients with AF have a higher mortality rate than those without AF. The prognostic significance of AF changes with age and gender while CHF is associated with the greatest risk of death.


Heavy metal resistance: a new role for P-glycoproteins in Leishmania.
P-glycoproteins are responsible for multidrug resistance in tumor cell lines and are thought to have a physiologic role in exporting cellular metabolites. We now report that a P-glycoprotein gene in the H region of the trypanosomatid protozoan Leishmania confers resistance to heavy metals when present in multiple copies. The Leishmania H region is frequently amplified in drug-resistant lines and is associated with metal resistance. Leishmania expression vectors were used to introduce multiple copies of segments of the Leishmania major H region into wild-type L. major promastigotes. Only constructs bearing a segment of L. major DNA containing the P-glycoprotein lmpgpA conferred arsenite resistance. Deletional analysis of the arsenite-resistant construct mapped resistance to the lmpgpA protein coding region. Lines expressing lmpgpA showed resistance to arsenite and trivalent antimonials, but not to pentavalent antimonials, zinc, cadmium, or the typical multidrug-resistant P-glycoprotein substrates vinblastine and puromycin. Transfection of the Leishmania tarentolae P-glycoprotein homologue ltpgpA resulted in a similar resistance profile. Thus, these pgpAs represent a functionally distinct group of P-glycoproteins which exhibit a substrate specificity similar to prokaryotic heavy metal pumps. Additionally, several arguments suggest that pgpAs may play a role in the susceptibility of Leishmania to clinically utilized antimonials.


[Communication competence in ultrasound examination in pregnancy].
293 physicians in the German-speaking part of Switzerland were questioned about their practices of providing information during prenatal ultrasound examinations. Physicians were asked both how they provide information about the examination procedure itself as well as how they inform the pregnant woman if a fetal anomaly is detected. In January 2000, a questionnaire was mailed to a sample of 500 physicians with a specialized qualification in pregnancy ultrasound examinations. The response rate was 59.2%. 90.1% of the physicians reported that they offer information consultations for routine ultrasound examinations. 50.2% mentioned in their answers to an open question that the screening for fetal anomalies is a component of their consultation. Female physicians mentioned this aspect significantly more often than male physicians. Regarding difficulties in the communication of a diagnosis of fetal malformation, the physicians reported most frequently the excessive strain on the pregnant woman, but also their own feelings of helplessness. The subjective emotional stress was considered as high; female physicians with experience of motherhood reported the highest levels of stress. Screening for fetal anomalies is a difficult and often neglected subject when providing information on prenatal ultrasound examinations. The communication of the diagnosis of a fetal anomaly is not only emotionally stressful for the parents, but also for the physicians and requires high levels of communication competence. These communication aspects should be considered in the training for ultrasound examinations during pregnancy.


Indeterminate pulmonary nodule in lung allograft characterized using dual-energy computed tomography.
Pulmonary nodules (PNs) arising in the lung transplant recipient pose a diagnostic challenge for providers. Conventional computed tomography (CT) has improved our ability to detect PNs in this population, but establishing a confident diagnosis with imaging alone remains difficult. Dual-energy spectral detector CT is a novel, emerging technology that provides insight into the radiographic behavior of PNs, and has potential in differentiating benign from malignant morphologies. Herein, we report a case of a PN in a lung transplant recipient whose initial diagnostic work-up was inconclusive, but then had the diagnosis rendered using a spectral detector CT.


Substantial interreader agreement for biopsy with reduction in biopsy rate: A multireader diagnostic performance study of ACR TI-RADS.
To assess diagnostic performance of ACR TI-RADS in thyroid cancer detection and evaluate interobserver agreement among radiologists with lower interpreting experience. Four radiologists retrospectively assessed 295 biopsied thyroid nodules from ultrasound studies performed between 2009 and 2019, blinded to histopathology. Diagnostic performance for cancer detection was determined individually, and interobserver agreement among four readers was evaluated with Fleiss kappa coefficient (ⱪ). 245 (83.1%) benign and 50 (16.9%) malignant nodules were evaluated. Diagnostic performance based on final TR level was consistent and without significant difference among four readers, with excellent sensitivity (≥98.0%) and negative predictive value (NPV) [≥94.4%] for TR levels 3 to 5. Diagnostic performance based on recommendation to biopsy has moderate sensitivity (≥62%) and high NPV (≥84.7%). Retrospective scoring with established ACR TI-RADS criteria would have substantially reduced the number of biopsies performed, with 63.2% of malignancy not biopsied meeting criteria for sonographic surveillance. Interobserver agreement on TI-RADS scoring for final TR level was fair (ⱪ = 0.39, 95% CI 0.32, 0.47), with substantial agreement for recommendation to biopsy (ⱪ = 0.64, 95% CI 0.58, 0.70). Substantial reduction in biopsy rate (up to 48%) would have been achieved using the ACR TI-RADS criteria, with 63% of malignancy not biopsied meeting criteria for sonographic surveillance. Interobserver agreement was fair for TI-RADS level scoring and substantial for recommendation to biopsy.


Binding of anti-acetylcholine receptor antibodies inhibits the acetylcholine receptor mediated cation flux.
A fast kinetics, spectroscopic technique has been applied to the study of the transient cation flux associated to the binding of cholinergic agonist to native acetylcholine receptor (AcChR)-rich membrane vesicles in presence of anti-AcChR antibodies. The technique is based on the collisional quenching of an intravesicularly trapped fluorophore by externally added T1+ which substitutes for physiologically occurring cations. Presence of polyclonal Fab fragments from goat anti-AcChR antibodies bound to the membrane AcChR promotes a 80-90% inhibition on the observed rate constants of T1+ influx. The observed inhibition process appears to follow a non-competitive pattern between antibody and cholinergic ligand binding, suggesting that in the AcChR protein the antigenic sites responsible for ion translocation may be other than those involved in ligand binding.


An analysis of QTc prolongation with atypical antipsychotic medications and selective serotonin reuptake inhibitors using a large ECG record database.
This study evaluated the effects of atypical antipsychotic drugs and selective serotonin reuptake inhibitors (SSRIs) on the corrected QT (QTc) interval using a large database obtained from clinical settings. Additionally, the effects of factors including age on QTc intervals were estimated. Using an open-access QT database (ECG-ViEW), QTc-lengthening effects of 14 selected atypical antipsychotics and SSRIs were compared to those of a positive control drug, cilostazol, and a negative control drug, diazepam. We also evaluated effects of age, sexgender, and select electrolyte levels on observed QTc intervals. The frequency of QTc prolongation with the pooled data of the 14 study drugs was lower than that with cilostazol (age-adjusted odds ratio (OR) = 0.43, 95% confidence interval (CI) = 0.27-0.69), but no significant difference was found relative to when compared with that with diazepam (age-adjusted OR = 0.89, 95% CI = 0.55-1.47). Furthermore, administration of the 14 study drugs significantly increased the QTc interval by 2.89 ms after each 10-year age increment (p-value < 0.0001). This study suggests that atypical antipsychotic drugs and SSRIs are less likely to be associated with QTc prolongation in clinical settings. In addition, age showed a significant association with the QTc interval. Further studies with well-characterized cohorts are warranted.


AIDS in an infant causing severe failure to thrive.
A nine-month-old female Jamaican infant presented with a history of severe failure to thrive, recurrent pneumonia and developmental delay. She was found to have hepatosplenomegaly, generalised lymphadenopathy and hypotonia. Investigations revealed polyclonal hypergammaglobulinaemia, cytomegalovirus in her urine, and patchy lung infiltrates on her chest radiographs. Three separate tests were positive for human immunodeficiency virus in both the infant and her mother, suggesting vertical transmission, and confirming AIDS as the cause of the severe failure to thrive.


Acute viral hepatitis presenting as cytomegalovirus, hepatitis E and Epstein-Barr virus IgM antibody positive.
We report a case of acute cytomegalovirus (CMV) infection with positive HEV and Epstein-Barr virus (EBV) serology. No patients have been reported positive for immunoglobulin (Ig)M antibodies to all three viruses. This patient had progressively increasing titres of IgM antibody for CMV, HEV and EBV. Only CMV DNA was detectable before antiviral treatment. After antiviral treatment, the patient recovered completely. At day 180 the CMV IgG test had converted to positive with CMV IgM (+), EBV IgM (-) and HEV IgM (-). Our report indicates that dependence upon serology alone is unreliable in the diagnosis of acute CMV, EBV and HEV infections. The diagnosis of CMV, HEV and EBV should be based on a combination of clinical features, serology and confirmatory PCR testing.


Baghel Prakash Prognosis Scoring System - Conceptual framework and preliminary psychometric validation.
The assessment of prognosis in a patient using a rigorous methodology is documented in old Ayurveda texts. But in present times, practitioners employ varied clinical assessment methods, which results in low inter-observer agreement. This study proposes a conceptual framework for developing a prognosis scoring system, which we refer to as the "Baghel Prakash prognosis scoring system" (BPPSS), and presents its preliminary psychometric validation. Classics of Ayurveda were extensively reviewed for item generation. Thirty-nine items were pooled initially. The Content Validity Index for Items (I-CVI), Scale-level Content Validity Index (S-CVI), and Content Validity Ratio (CVR) were calculated. The final framework contained 30 items. The scoring system was pilot-tested in a sample of 30 patients with various disease conditions. The reliability checks were done to evaluate internal consistency. Inter-item correlations, item-total correlations, and Cronbach's alpha (if an item is deleted) were used to conduct item analysis for the instrument. The framework developed includes a set of early clinical warning signs and a prognosis scoring system of 30 items with a provision for clinical adjustment of scores for selected items. Internal consistency of the scoring system during the initial psychometric validation revealed a value of .761 for Cronbach's alpha (based on standardized items), and item analysis revealed that most of the items had acceptable correlation coefficients between .3 and .7. Based on this preliminary validation study, we found that this prognosis framework, based on the principles of Ayurveda, may be used in varied clinical situations. The preliminary psychometric validation experiments yielded satisfactory results. The framework also has potential in clinical research, such as selecting patients with similar prognostic scores for comparability in case and control groups of clinical trials. This tool can be utilized as a reliable outcome measure after conducting enough validation studies.


Effect of highway directional signs on driver mental workload and behavior using eye movement and brain wave.
The rapid development of expressways has led to an increasing number of place names that must be displayed on road guide signs. As a result, multi-board guide signs have been increasingly set up on expressways. The main aim of this study was to analyze the effect of the directional road sign displayed on multi and single-board signs on driver mental workload and behavior. 32 participants including 16 females (mean age = 24.7 years, standard deviation = 1.9 years) participated in the experiment and completed 3 driving simulation scenes. The setting of each scene-sign board was different: 1 board, 2 boards, and 3 boards. The driver needed to reach the designated destination according to the guidance of the road signs. Eye tracker was used to measure the fixation, saccade and electroencephalogram (EEG) was used to measure the alpha (8-13 hz) band absolute power in different signage scenarios. There are two major findings of the study. First, when the number of place names is less than or equal to 7, the multi-board sign generates more mental workload than the single-board sign does. The alpha band power of the driver's frontal area under the multiple boards is lower and affects driving performance (the deceleration is greater). Second, when the number of place names is more than 7, there is no significant difference in the effect on mental workload whether multi or single-board sign is used. However, compared to the single-board sign, drivers in the case of multi-board sign are likely to reduce the fixation duration and increase the number of saccades. The results suggest that it is not necessary to use multi-board signs when the number of place names is less than 7. These findings provide more safety considerations for the setting of multi-board guide signs in the future.


Protein, leucine aminopeptidase, esterase, acid phosphatase and photosynthetic responses of oleander (Nerium oleander L.) during cold acclimation and freezing treatments.
Six-month-old oleander (Nerium oleander L.) pot plants, derived from vegetative propagation by cuttings, were tested for their ability to cold hardening. Damage of the non-acclimated (NA) plants was visible when treated by low freezing temperatures (below -2 degrees C). The responses of total proteins, leucine aminopeptidase (LAP), esterase (EST) and acid phosphatase (ACP) isoforms of NA and cold-acclimated (CA; 4 degrees C for 14 days) plants were compared using polyacrylamide gel electrophoresis. These molecular markers were also compared in NA and CA plants which received for 2h temperatures of 0, -2, -4, -6 and -8 degrees C. A new 38-kDa polypeptide appeared from day 7 to 14 during the acclimation treatment in the bark extracts and on day 14 in the leaf extracts. The above-mentioned polypeptide band (38 kDa) strongly appeared in all freezing treatments (0, -2, -4, -6 and -8 degrees C) in both bark and leaf extracts of the CA plants. Alterations in the number and the intensity of LAP and EST isoforms as well as in the intensity of ACP isoforms were observed in both bark and leaf of the CA oleander plants. A newly expressed EST isoform is proposed as biochemical marker for the cold acclimation treatment. CO2 assimilation rates (A) as well as transpiration rates (E) in NA plants were positive in 0 degrees C and negative in all temperatures below zero in the freezing treatments. In contrast, CO2 assimilation rates (A) and transpiration rates (E) were positive in CA plants in all temperatures of freezing treatment. A significant decrease (P<0.05) in chlorophyll (Chl) a, Chl a+b concentration and Chl a/b ratio were noticed in oleander plants during the acclimation treatment (from day 0 to 14), while Chl b concentration was unchanged at the respective time. On the other hand, no significant (P<0.05) differences were observed in the freezing treatments.


Sociocultural factors related to the physical activity in boys and girls: PeNSE 2012.
To verify in male and female Brazilian adolescents the association of demographic, psychosocial, behavioral and sociocultural characteristics with the regular practice of physical activity. The sample consisted of 109,104 adolescents from all Brazilian states attending the 9th year of elementary education in 2012. The response variable was the regular practice of physical activity (300+ minutes/week). The explanatory variables were grouped into four fields: demographic, psychosocial, behavioral and sociocultural. The Poisson regression was stratified by sex to evaluate the association. The prevalence of active adolescents was 20.2%, higher in boys (27.9%) than in girls (13.1%). It was observed a greater practice of physical activity in boys of lower age group, children of mothers with higher schooling, who consumed healthy foods such as beans, fruits, vegetables, and milk, as well as among those with family supervision. At the same time, unhealthy habits such as insomnia and alcohol consumption were also positively associated with physical activity. In girls, greater physical activity was observed among those who lived with mothers and whose mothers had higher schooling. In addition to family supervision, the practice of physical activity in girls was also positively associated with the frequency of meals with their parents. However, as in boys, insomnia and alcohol consumption were associated with an increase in the practice of physical activity. One-fifth of adolescents practice physical activity regularly, demonstrating the need for specific public policies to increase the percentage of active young people in the country. Maternal schooling, healthy eating habits and family supervision were associated with regular physical activity in boys and girls, evidencing the importance of the family for the acquisition of healthy habits in this age group.


Mastitis, Breast Abscess, and Granulomatous Mastitis.
Breastfeeding is immunoprotective and World Health Organization recommends exclusive breastfeeding for about six months with continuation of breastfeeding for one year or longer as mutually desired by mother and infant. But the target for duration of exclusive breastfeeding has not been reached in a significant number of women. It may be due to inflammatory breast disease such as milk stasis or lactational mastitis.In this chapter we discuss the most common complications of breastfeeding including milk stasis, mastitis, and breast abscess. Also idiopathic granulomatous mastitis, a less common condition, is discussed due to its confusing characteristics and not universally-accepted treatment strategies .Breastfeeding mastitis is inflammation of the breast that can be infectious or non-infectious. With proper diagnosis and treatment of this condition, more severe complications like breast abscess could be avoided, so that breastfeeding could be continued in some circumstances.


[Roentgeno-morphological pulmonary changes following bleomycin therapy].
100 patients with squamous cell carcinoma of the head and neck, who underwent a bleomycin therapy pre- and postoperative have been followed up retrospectively for radiological signs of bleomycin-typical side-effects of the lung. The frequency of such changes was 24% and there were 2 peaks after 1 week and 4-6 weeks from the start of therapy. An increased risk to develop an interstitial exsudation and fibrosis seems to exist only for patients with preexistent emphysema of the lung.


A turbidometric assay for measuring proteins in culture media.
An immunoturbidometric assay was developed for the measurement of proteins in culture fluids of hepatocytes. The assay is simple to perform and avoids the biohazards associated with radioimmunoassays. The limit of detection of this assay exemplified by hemopexin and transferrin is 5 ng/ml protein. This degree of sensitivity is attained by incorporating into the procedure the addition of polyethylene glycol to enhance formation of primary antigen-antibody complexes and of a second antibody to further increase the immune complex size, which favors the ratio of specific to background light scattering.


Coupling motion between rearfoot and hip and knee joints during walking and single-leg landing.
The objective of the current study was to investigate the kinematic relationships between the rearfoot and hip/knee joint during walking and single-leg landing. Kinematics of the rearfoot relative to the shank, knee and hip joints during walking and single-leg landing were analyzed in 22 healthy university students. Kinematic relationships between two types of angular data were assessed by zero-lag cross-correlation coefficients and coupling angles, and were compared between joints and between tasks. During walking, rearfoot eversion/inversion and external/internal rotation were strongly correlated with hip adduction/abduction (R=0.69 and R=0.84), whereas correlations with knee kinematics were not strong (R≤0.51) and varied between subjects. The correlations with hip adduction/abduction were stronger than those with knee kinematics (P<0.001). Most coefficients during single-leg landing were strong (R≥0.70), and greater than those during walking (P<0.001). Coupling angles indicated that hip motion relative to rearfoot motion was greater than knee motion relative to rearfoot motion during both tasks (P<0.001). Interventions to control rearfoot kinematics may affect hip kinematics during dynamic tasks. The coupling motion between the rearfoot and hip/knee joints, especially in the knee, should be considered individually.


Circulating Soluble CD163 is Associated with Steatohepatitis and Advanced Fibrosis in Nonalcoholic Fatty Liver Disease.
Soluble CD163 (sCD163), a marker of Kupffer cell activation detectable in serum, correlates with inflammation and fibrosis in chronic viral hepatitis, but its role in nonalcoholic fatty liver disease is unknown. We hypothesized that sCD163 would correlate with nonalcoholic fatty liver disease activity and fibrosis. Liver biopsies and serum were obtained from 145 obese subjects undergoing gastric bypass surgery. Subjects were divided into four groups based on fibrosis stage and nonalcoholic fatty liver disease activity score (NAS); Group 1: F0, NAS=0; Group 2: F<2, 0<NAS<5; Group 3: NAS≥5, F<3; or Group 4: F≥3, any NAS. Serum sCD163 and the monocyte/macrophage marker sCD14 were measured by enzyme-linked immunosorbent assay. Relationships between sCD163, sCD14, fibrosis stage, and NAS were examined. Area under the receiver operating charateristic for the diagnosis of nonalcoholic steatohepatitis based on the Clinical Research Network definition was calculated. sCD163 increased with progressive liver histology, with lowest values in normal histology and highest levels in those with nonalcoholic steatohepatitis and advanced fibrosis (Group 1: 552 ng/ml, Group 2: 721 ng/ml, Group 3: 803 ng/ml, and Group 4:1,031; P=0.001). sCD14 also differed significantly across groups (Group 1: 1,877 ng/ml, Group 2: 1632 ng/ml, Group 3: 1,706 ng/ml, and Group 4: 2111; P=0.008, respectively). sCD163 correlated with steatosis grade (P<0.001), lobular inflammation (P=0.033), and hepatocyte ballooning (P<0.001). In a multivariable ordered logistic regression model, there was a significant association between every 100 ng/ml increase in sCD163 and higher fibrosis stage, with an odds ratio of 1.16 (95% confidence interval 1.02-1.31), P=0.020. The odds ratios of the association between every 100 ng/ml increase in sCD163 and higher NAS was 1.17 (95% confidence interval 1.04-1.32), P=0.010. A sCD163-based predictive score demonstrated an area under the receiver operating charateristic of 0.70 (95% confidence interval: 0.58-0.82) for the diagnosis of nonalcoholic steatohepatitis. Soluble CD14 did not correlate with fibrosis stage or NAS. In obese subjects, serum sCD163, but not sCD14, correlated with fibrosis stage and NAS. These data support a role for activated Kupffer cells in the pathogenesis of nonalcoholic steatohepatitis and fibrosis, and suggest potential clinical utility for assessment of sCD163 levels.


N6-isopentenyladenosine induces cell death through necroptosis in human glioblastoma cells.
Targeting necroptosis is considered a promising therapeutic strategy in cancer, including Glioblastoma Multiforme (GBM), one of the most lethal brain tumors. Necroptosis is a mechanism of programmed cell death overcoming the apoptosis resistance mechanism underlying GBM tumorigenesis and malignant progression. N6-isopentenyladenosine (iPA), adenosine modified with isoprenoid derivative, displays antitumor activity in different cancer models. In previous studies, we demonstrated that iPA interferes with EGFR signaling reducing glioma cell viability. Here, we show that iPA induces necroptosis in glioblastoma cell lines and in primary cells established from tumor explants, without affecting the viability of non-cancerous brain cell lines, (Normal Human Astrocyte). The activation of RIP1, RIP3, and MLKL and the upregulation of necrosome formation were increased upon iPA treatment while caspase-3, caspase-8, and PARP were not activated in GBM cells. Co-treatment with specific necroptosis inhibitor necrostatin-1 (Nec-1) or Necrosulfonamide (NSA) prevented cell death caused by iPA treatment while the general caspase inhibitor Z-VAD-fluoromethylketone (z-VAD-fmk) did not elicit any effect, suggesting that this molecule induces caspase-independent necroptosis. These results suggest that iPA treatment can be able to bypass the apoptosis resistance mechanism in glioblastoma thereby offering higher therapeutic efficacy.