One-step separation and purification of three lignans and one flavonol from Sinopodophyllum emodi by medium-pressure liquid chromatography and high-speed counter-current chromatography.
Lignans and flavonols are the primary constituents of Sinopodophyllum emodi and have been used as cathartic, anthelmintic, chemotherapeutic and anti-hypertensive compounds. Although these compounds have been isolated, there have been no reports on the separation of 4'-demethyl podophyllotoxin, podophyllotoxin, deoxypodophyllotoxin and kaempferol in one step by medium-pressure liquid chromatography (MPLC) and high-speed counter-current chromatography (HSCCC). Development of an efficient method for the preparative separation and purification of three lignans and one flavonol from S. emodi. The precipitate of crude extracts was first separated by MPLC into four parts, numbered GJ-1, GJ-2, GJ-3 and GJ-4. GJ-1 was separated and purified by HSCCC using a solvent system composed of n-hexane:ethyl acetate:methanol:water (1.75:1.5:1:0.75, v/v/v/v). The purities of the target compounds were assessed using high-performance liquid chromatography (HPLC) and chemical structures were identified by (1) H-NMR and (13) C-NMR. The HSCCC and MPLC methods were successfully used for the preparative separation and purification of 4'-demethyl podophyllotoxin (8.5 mg, 92.4%), podophyllotoxin (40.1 mg, 92.1%), deoxypodophyllotoxin (4.6 mg, 98.1%), and kaempferol (1.6 mg, 96.7%) from a 100 mg sample. Three lignans (4'-demethyl podophyllotoxin, podophyllotoxin, deoxypodophyllotoxin) and one flavonol (kaempferol) were successfully isolated by HSCCC and MPLC in one step.


Evaluation of dendrimer type bio-reducible polymer as a siRNA delivery carrier for cancer therapy.
Small interfering ribonucleic acid (siRNA), 20-25 base pairs in length, can interfere with the expression of specific genes. Recently, many groups reported the therapeutic intervention of siRNA in various cancer cells. In this study, dendrimer type bio-reducible polymer (PAM-ABP) which was synthesized using arginine grafted bio-reducible poly(cystaminebisacrylamide-diaminohexane) (ABP) and polyamidoamine (PAMAM) was used to deliver anti-VEGF siRNA into cancer cell lines including human hepatocarcinoma (Huh-7), human lung adenocarcinoma (A549), and human fibrosarcoma (HT1080) cells and access their potential as a siRNA delivery carrier for cancer therapy. PAM-ABP and siRNA formed polyplexes with average diameter of 116 nm and charge of around +24.6 mV. The siRNA in the PAM-ABP/siRNA polyplex was released by 5mM DTT and heparin. VEGF gene silencing efficiency of PAM-ABP/siRNA polyplexes was shown to be more effective than PEI/siRNA polyplexes in three cell lines with the following order HT1080>A549>Huh-7.


The fine structure of human digital arterio-venous anastomoses (Hoyer-Grosser's organs).
Hoyer-Grosser's organs were studied in human digital biopsies. The fine structure of both the supplying arteries and collecting veins was found to be inconspicuous. Endothelial cells in the AV canals form a continuous layer. They are characterized by their rich content of specific organelles (Weibel-Palade bodies), especially in the venous segments. The epitheloid zone is composed of a variety of ramified smooth muscle cells (RSM). These appear either dense, when well provided with bundles of myofilaments, or clear, when including only a few myofilaments. The nuclei of dense RSM show condensed chromatin, while those of clear RSM are larger with loose chromatin texture. In addition, all transitional forms occur. Cell organelles are rarely seen within all types of RSM. The cytoplasmic processes reach other as well as endothelial cells. The preservation of our material did not allow observation of specialized membrane contacts in these zones. All RSM are invested with a regular basal lamina and well provided with surface vesicles. Profiles of free basal lamina material and faint collagen (argyrophil) fibers are seen in the wide intercellular spaces. RSM poor in myofilaments are interpreted to represent "epitheloid cells" of light microscopy. Their number constantly decreases from the arterial segment of the AV canal to the venous segment. Here the cytoplasmic processes of RSM become less pronounced and the wall of the anastomotic segment continuously changes to that of the collecting vein. Dense RSM rich in myofilaments are compared with pacemaker cells found inthe ureter. Both internal and external elastic membranes are absent in AV canals. A dense network of unmyelinated nerve fibers is found in the adventitial layer of the AV canal, especially in its arterial segment. The axons branch out from small dermal nerves which also contain two or three myelinated axons. The myelin sheaths terminate where the axons reach the adventitia of the AV canals. Axon varicosities filled with mitochondria are thought to be terminals of myelinated axons and are interpreted as receptory. Axon varicosities with synaptic-type vesicles are assumed to be terminals of sympathetic and para sympathetic fibers. All axon profiles are confined to the adventitial layer of the anastomotic segment.


Finding smORFs: getting closer.
Millions of small open reading frames exist in eukaryotes. We do not know how many, or which are translated, but bioinformatics is getting us closer to the answer. See related Research article: http://www.genomebiology.com/2015/16/1/179.


Continuous flow-extractive desorption electrospray ionization: analysis from "non-electrospray ionization-friendly" solvents and related mechanism.
Due to their low polarities and dielectric constants, analytes in solvents such as hexane, chloroform, and ethyl acetate exhibit poor electrospray ionization (ESI) efficiency. These are deemed to be "non-ESI-friendly" solvents. Continuous flow extractive desorption electrospray ionization (CF-EDESI) is a novel ambient ionization technique that was recently developed in our group to manipulate protein charge distributions. Here we demonstrate its potential for ionizing analytes from non-ESI-friendly solvents. This feature makes CF-EDESI attractive to the general analytical community due to its apparent potential in lipidomics, normal phase separations, and hyphenation of mass spectrometry with HPLC-NMR systems. In this context, interest was subsequently initiated to discern mechanistic aspects of CF-EDESI. To achieve this, mechanistic experiments associated with a seemingly similar ambient ionization technique, extractive electrospray ionization (EESI), were emulated to compare CF-EDESI and EESI. Analysis of a series of fatty acids in multiple solvents in the negative ionization mode revealed differences between the two techniques. Whereas EESI has been previously shown to operate via extraction of analytes into the spray solvent, data presented here for CF-EDESI point toward a liquid-liquid mixing process to facilitate ionization. Further, a partial factorial design experiment was performed to evaluate the effects of different experimental variables on signal intensity. Sample flow rate was confirmed to be among the most significant factors to affect sensitivity. As a whole, the work presented provides greater insight into a new ambient ionization process, which exhibits expanded capabilities over conventional ESI; in this case, for direct analysis from non-ESI-friendly solvents.


Riedel's thyroiditis: clinical, pathological and imaging features.
Riedel's thyroiditis (RT) is an extremely rare form of chronic thyroiditis, characterised by a fibroinflammatory process which partially destroys the thyroid, often involving surrounding tissues. We describe an unusual case of RT in a 38-year-old woman, and discuss its typical clinical, pathological, ultrasound, radionuclide scanning and magnetic resonance findings. We conclude that the diagnosis of RT is highly suggestive in the presence of a hard gland (or nodule), fixed to adjacent structures; 'cold' on Tc99 scan; hypoechoic with absence of vascular flow, invading the adjacent neck structures on ultrasound; hypointense in T1- and T2-weighted MR images; and showing fibrous tissue fragments with inflammatory cells at cytological examination. Thyroidectomy has to be performed to confirm the diagnosis and to relieve pressure symptoms.


Agricultural trade impacts global phosphorus use and partial productivity.
The spatio-temporal distribution, flow and end use of phosphorus (P) embedded in traded agricultural products are poorly understood. Here we use global trade matrices to analyse the partial factor productivity of P (output per unit of P input) for crop and livestock products in 200 countries and their cumulative contributions to the export or import of agricultural products over 1961-2019. In these six decades, the trade of agricultural P products has increased global partial factor productivity for crop and livestock production and has theoretically saved 67 Tg P in fertilizers and 1.6 Tg P in feed. However, trade is now at risk of contributing to wasteful use of P resources globally due to a decline in trade optimality, as agricultural products are increasingly exported from low to high partial factor productivity countries and due to P embedded in imported agricultural products mainly lost to the environment without recycling. Integrated crop-livestock production systems and P-recycling technologies can help.


Does early communication mediate the relationship between motor ability and social function in children with cerebral palsy?
Children diagnosed with neurodevelopmental conditions such as cerebral palsy (CP) are at risk of experiencing restrictions in social activities negatively impacting their subsequent social functioning. Research has identified motor and communication ability as being unique determinants of social function capabilities in children with CP, to date, no research has investigated whether communication is a mediator of the relationship between motor ability and social functioning. To investigate whether early communication ability at 24 months corrected age (ca.) mediates the relationship between early motor ability at 24 months ca. and later social development at 60 months ca. in a cohort of children diagnosed with cerebral palsy (CP). A cohort of 71 children (43 male) diagnosed with CP (GMFCS I=24, 33.8%, II=9, 12.7%, III=12, 16.9%, IV=10, 14.1%, V=16, 22.5%) were assessed at 24 and 60 months ca. Assessments included the Gross Motor Function Measure (GMFM), the Communication and Symbolic Behaviour Scales-Developmental Profile (CSBS-DP) Infant-Toddler Checklist and the Paediatric Evaluation of Disability Inventory (PEDI). A mediation model was examined using bootstrapping. Early communication skills mediated the relationship between early motor abilities and later social functioning, b=0.24 (95% CI=0.08-0.43 and the mediation model was significant, F (2, 68)=32.77, p<0.001, R(2)=0.49. Early communication ability partially mediates the relationship between early motor ability and later social function in children with CP. This demonstrates the important role of early communication in ongoing social development. Early identification of communication delay and enriched language exposure is crucial in this population.


The deazaguanine-derivative, queuine, affects cell proliferation, protein phosphorylation and the expression of the proto oncogenes c-fos and c-myc in HeLa cells.
In higher eukaryotes the hypermodified guanine analogue, queuine (7-(5-[(1S,4S,5R)-4,5-dihydroxy-2-cyclopentene-1-yl) amino)-methyl)-7-deazaguanine), occurs free or as modified nucleoside (Q) in the anticodon of specific tRNAs. Fast proliferating tissues and tumors contain considerable amounts of free queuine and Q-deficient tRNAs. Here we show that HeLa cells can be grown in the absence or presence of queuine. In response to queuine, and under appropriate conditions, (i) the proliferation of HeLa cells was stimulated, (ii) the steady-state level of c-fos mRNA was reduced, contrary that of c-myc mRNA was elevated, and (iii) in cytosolic extracts protein phosphorylation especially for a 42 kDa protein was significantly increased. The results suggest that queuine substitutes for growth factors in a signal transduction pathway.


Effect of antrectomy and drug-induced achlorhydria on urinary excretion of N-terminal big gastrin immunoreactivity in rats.
Immunoreactivities of urinary N-terminal big gastrin and serum C-terminal gastrin were determined in intact and antrectomized rats by radioimmunoassay using two antisera specific for N- and C-termini of big gastrin, respectively. Gel filtration of urine extract from intact rat showed a single giant peak of N-terminal big gastrin immunoreactivity eluted in a later position than 1-17 gastrin-34, indicating that N-terminal peptides smaller than 1-17 gastrin-34 are excreted in urine. Serum C-terminal gastrin concentration in antrectomized rats was about one sixth that in intact rats. Urinary excretion of N-terminal big gastrin in antrectomized rats was about one sixth that in intact rats. 2 week treatment with E3810, a proton pump inhibitor, (40 mg/kg/day, s.c.) induced urinary excretion of N-terminal big gastrin in parallel with a marked increase in serum C-terminal gastrin concentration in intact rats. Antrectomy completely prevented both the increase in urinary excretion of N-terminal big gastrin and the elevation of serum C-terminal gastrin induced by administration of E3810. There was an excellent correlation between serum concentration of C-terminal gastrin and urinary excretion of N-terminal big gastrin. These results suggest that urinary N-terminal big gastrin, which mostly originates from the gastric antrum, is a useful indicator of gastrin secretion in the rat.


Exophthalmometry values of Turkish adult population and the effect of age, sex, refractive status, and Hertel base values on Hertel readings.
To establish normal exophthalmometry values in the adult Turkish population, and the impact of age, gender, interpupillary distance, Hertel base selection, and refractive status on globe position. Exophthalmometry measurements and refractive status of 2477 subjects were conducted in one tertiary and five primary health care centers. Change of globe position by age, intersex differences in terms of globe position, correlations of Hertel base with exophthalmometry results, and interpupillary distance (IPD) were evaluated. Multiple linear regression analysis was performed to test determination effect of each variable on final Hertel reading. Median Hertel reading was 13 mm, and 95% of the population had an upper limit of 17 mm for both eyes. There was a negative correlation between spherical equivalent of refractive status and exophthalmometry results and a weak positive correlation between IPD and exophthalmometry result. Mean Hertel value was found to decrease significantly after the third decade. Hertel base value was found to have moderate linear correlation with Hertel results. A weak correlation was detected with Hertel base/IPD ratio with final Hertel results. Multiple linear regression analysis was performed and only 13% and 20% of change in Hertel values bilaterally were found to be determined by other variables (age, IPD, refractive status, and Hertel base value) for females and males. Normative dataset for exophthalmometry results of the Turkish population is established to be used in clinical practice and research. Only 13% to 20% of change in Hertel values was detected to be determined by age, IPD and Hertel base values.


Water activity of poultry litter: Relationship to moisture content during a grow-out.
Poultry grown on litter floors are in contact with their own waste products. The waste material needs to be carefully managed to reduce food safety risks and to provide conditions that are comfortable and safe for the birds. Water activity (Aw) is an important thermodynamic property that has been shown to be more closely related to microbial, chemical and physical properties of natural products than moisture content. In poultry litter, Aw is relevant for understanding microbial activity; litter handling and rheological properties; and relationships between in-shed relative humidity and litter moisture content. We measured the Aw of poultry litter collected throughout a meat chicken grow-out (from fresh pine shavings bedding material to day 52) and over a range of litter moisture content (10-60%). The Aw increased non-linearly from 0.71 to 1.0, and reached a value of 0.95 when litter moisture content was only 22-33%. Accumulation of manure during the grow-out reduced Aw for the same moisture content. These results are relevant for making decisions regarding litter re-use in multiple grow-outs as well as setting targets for litter moisture content to minimise odour, microbial risks and to ensure necessary litter physical conditions are maintained during a grow-out. Methods to predict Aw in poultry litter from moisture content are proposed.


Isolation and characterization of mouse CD7 cDNA.
The human CD7 antigen is a glycoprotein, M(r) 40,000, expressed on the surface of peripheral blood T-lymphocytes and thymocytes, and is the earliest surface antigen to appear on T-cell lineage cells. In this study, putative mouse CD7 cDNA was identified based on its similarities with human CD7. Five independent clones originating from the same mRNA species were isolated (designated as mCD7) by screening a mouse thymocyte cDNA library with human CD7 cDNA, J61, under moderate stringency. The longest insert of a 995 base pair had an open reading frame of 210 amino acids. Northern blot analysis using the mouse CD7 cDNA probe demonstrated a single 1.2 kilobase mRNA in the thymus, spleen, bone marrow, and small intestine. The protein deduced from mCD7 cDNA consisted of the leader, extracellular, transmembrane, and cytoplasmic domains of 24, 126, 21, and 39 amino acids, respectively, based on the hydrophobicity plot and the structure of human CD7. The extracellular domain contained three potential N-glycosilation sites, while the cytoplasmic domain contained one potential protein kinase C phosphorylation site. The amino acid sequence had 45.5% similarity with human CD7, while the similarities for the individual domains ranged from 49.2% to 63.2%. The six highly conserved regions, which may possibly be involved with still unknown CD7-mediated functions, were located in the extracellular and cytoplasmic domains.


Efficient ultrafast laser writing with appropriate polarization.
Appropriate polarization utilization makes the electric field vector direction and the statistically oriented localized states suitable for enhancing light-matter interactions so as to improve the efficiency of ultrafast laser writing, which will remarkably reduce the pulse energy and increase the processing speed for high density optical data storage, as well as manufacturing three-dimensional integrated optics and geometric phase optical elements.


Hypothesis: hyperhomocysteinemia is an indicator of oxidant stress.
Elevated plasma homocysteine levels are associated with an increased risk of atherosclerosis and thrombosis, as well as a variety of other pathologies such as birth defects, Alzheimer's disease and other dementias, osteoporosis, diabetes and renal disease. Homocysteine metabolism is catalyzed by a number of enzymes that require B-vitamins as cofactors, and homocysteine levels are particularly responsive to folate status. The predictive power of plasma homocysteine level as a risk factor for atherothrombotic orders raised the appealing hypothesis that reduction of homocysteine levels by vitamin supplementation might result in a commensurate reduction is the risk of atherothrombotic events. Unfortunately, most clinical trials failed to show a significant benefit of vitamin supplementation on cardiovascular events, in spite of significant lowering of plasma homocysteine levels. Thus, it is not clear whether homocysteine actually plays a causal role in many pathologies with which it is associated, or whether it is instead a marker for some other underlying mechanism. A large body of data links hyperhomocysteinemia and folate status with oxidant stress. In this article I review data that suggests that homocysteine not only promotes cellular and protein injury via oxidant mechanisms, but is also a marker for the presence of pathological oxidant stress. Thus, it is possible that hyperhomocysteinemia is not a common primary cause of atherothrombotic disorders in the general population, but rather a marker of systemic or endothelial oxidant stress that is a major mediator of these disorders.


Ecological analyses of nesting success in evening grosbeaks.
We studied the nesting success of Evening Grosbeaks (Coccothraustes vespertinus) inhabiting two areas of the Front Range of the Rocky Mountains of Colorado from 1983-1987. Sixty-four nests were followed during building, incubating, brooding, and fledging; 54.7% were successful (young fledged). The largest number of nests failed during incubation. Nests started later were more successful than nests begun earlier in the season. Failure was most likely due to severe weather, abandonment during building, or predation. Specific habitat characteristics of grosbeak nesting sites and where nests were placed in trees were consistently associated with nesting success. Successful nests, when compared with nests that failed, were: (1) built in more open areas characterized by dispersed vegetation and a higher minimum canopy, (2) oriented in more southerly directions, (3) built closer to the main trunk of the nest tree, and (4) built in larger trees. Current ideas about whether or not birds actually select nest-sites are briefly discussed. We conclude that some grosbeaks "optimally select" nest sites where the likelihood of producing fledglings is higher than in other areas.


Monitoring of DNA damage in haemocytes of freshwater mussel Sinanodonta woodiana sampled from the Velika Morava River in Serbia with the comet assay.
This study was undertaken to investigate the potential of the freshwater mussel Sinanodonta woodiana for detection of genotoxic pollution of the environment. Study was performed at two sites in the Velika Morava River, from May 2010 to February 2011. The alkaline comet assay on haemocytes was used, and the olive tail moment (OTM) was chosen as a measure of DNA damage. The specimens held on acclimation under controlled laboratory conditions for 10d were used as a control. Chemical analysis revealed the presence of phosphates and increased concentrations of zinc, copper and nickel at both sites during the entire sampling period. The values of OTM in mussels collected from the environment, significantly correlated with the concentration of zinc (r=0.6248), temperature (r=0.7006) and dissolved oxygen (r=0.7738). Seasonal variations in genotoxic response were observed, with the highest OTM values obtained during summer months. Preliminary results of the in vitro study indicated the effect of water temperature on genotoxic response to zinc and cadmium in S. woodiana suggesting that the presence of genotoxic pollutants during months with lower temperature could be under-estimated. Obtained results indicate that S. woodiana could be a valuable tool for active biomonitoring of aquatic environments and emphasizes the importance of seasonal genotoxic monitoring with this organism.


Short- and medium-chain chlorinated paraffins in commercial complementary baby food produced in different European countries: Occurrence, congener group profiles, portion-based dietary intake, and risk assessment.
This study was performed to evaluate the presence of chlorinated paraffins (CPs) in complementary baby food from different European countries, to assess portion-based dietary intake of CPs, and to characterize possible risks to children arising from CPs uptake. In total, 86 baby food samples from 22 countries were grouped into eight food-type categories considering the WHO classification. In six out of eight sample categories, predominance of medium-chain CPs (MCCPs) concentrations over short-chain CPs (SCCPs) was observed and, contrary to expectations, samples that contained ingredients of animal origin (e.g., meat or fish) did not display elevated CPs levels. The median ∑CPs concentrations for selected sample categories ranged from 0.8 ng g-1 for desert to 2.8 ng g-1 found in dry cereal and porridge subgroups, while the highest CPs concentration of 387 ng g-1 was observed in dry porridge. Among the selected groups of baby foods, portion-based intake of ∑CPs varied from 1.0 to 43.6 ng kg-1 body weight (b.w.) while exposure estimations indicated that portion-based dietary exposure through consumption of complementary baby food was the highest for infants aged 3-12 months due to the lower body weight. The risk characterization according to the European Food Safety Authority (EFSA) approach showed that the margin of exposure (MOE) figures higher than the critical value for ∑CPs, indicating that the dietary exposures through consumption of complementary baby food are unlikely to be of significant health concern for children. However, it should be pointed out that the performed risk assessment covered only SCCPs and MCCPs, while the occurrence of other groups from CPs and CPs related compounds (e.g., very short-chain CPs, chlorinated alkenes, or long-chain CPs (LCCPs)) was not addressed.


Effects of zooplankton availability and foraging mode on cannibalism in three dragonfly larvae.
Cannibalism is likely to operate as a form of population control in dragonfly larvae. I performed aquarium experiments to investigate the effect of foraging activity and zooplankton availability on cannibalism in three dragonfly larvae. Large Cordulia aenea larvae showed low activity, and large Leucorrhinia dubia larvae showed high activity irrespective of zooplankton availability. In contrast, large Coenagrion hastulatum larvae changed from high activity in the absence to low activity in the presence of zooplankton. Small Cordulia aenea larvae were active in the absence of large conspecifics irrespective of zooplankton availability. In the presence of large conspecifics they showed a reduced activity when zooplankton were present. Small L. dubia larvae showed high activity and small Coenagrion hastulatum larvae low activity irrespective of pressence or absence of zooplankton and large conspecifics. In all three species cannibalism was highest in the absence of zooplankton. In the absence of zooplankton cannibalism was low in Coenagrion hastulatum compared to the other two species. On the contrary, in the presence of zooplankton, cannibalism did not differ between the three species.


Pressurized GRAS solvents for the green extraction of phenolic compounds from hibiscus sabdariffa calyces.
An environmental friendly extraction procedure has been tested to extract phenolic compounds from H. sabdariffa calyces using pressurized GRAS solvents. A central composite rotatable design (CCRD) was performed to evaluate the influence of the main operational conditions: temperature (40-200 °C) and solvent composition based on aqueous hidroalcoholic solutions (0-100% ethanol). Phenolic composition of experimental extracts analyzed by HPLC-ESI-QTOF-MS showed that higher temperatures and greater ethanol percentages drove to solvents with lower dielectric constants, which resulted in extracts with major quantities of phenolic compounds. An exception was the extraction of cyanidin-3-sambubioside that could only be quantified in extracts performed at the lowest temperature (40 °C) due to its thermal sensibility. In addition, a RSM was carried out with the aim to maximize the extraction of total phenolic content. To this end, the predicted optimal extraction conditions by RSM were 200 °C and 100% (v/v) of ethanol. Results showed that temperature and ethanol percentage had a significant influence on the extraction of total phenolic compounds (p value < 0.05). The mathematical model pointed out 200 °C of temperature and 100% of ethanol as the optimum conditions to perform the isolation of phenolic compounds by means of pressurized GRAS solvents.


Unprecedented Microbial Conversion of Biliverdin into Bilirubin-10-sulfonate.
Biliverdin (BV) possesses antioxidant and anti-inflammatory properties, with previous reports identifying protection against oxidant and inflammatory injury in animal models. Recent reports indicate that intra-duodenal administration of BV results in the formation of an uncharacterised metabolite, which is potently absorbed into the blood and excreted into the bile. This compound may be responsible for protection against inflammatory responses. This study aimed to identify novel, enterally-derived BV metabolites and determine the source of their metabolic transformation. Rat duodena and bacterial cultures of Citrobacter youngae were treated with BV and subsequently analysed via high performance liquid chromatography/high resolution tandem mass spectrometry to identify and characterise metabolites of BV. A highly abundant metabolite was detected in duodenal wash and bacterial culture supernatants with a 663.215 m/z (3 ppm mass accuracy) and a composition of C33N4O9H36S, which conformed to the predicted structure of bilirubin-10-sulfonate (BRS) and possessed a λmax of 440 nm. Bilirubin-10-sulfonate was then synthesized for comparative LCMS/MS analysis and matched with that of the biologically formed BV metabolite. This report confirms the formation of a previously undocumented metabolite of BV in mammals, indicating that a new metabolic pathway likely exists for BV metabolism requiring enteric bacteria, Citrobacter youngae. These data may have important implications with regard to understanding and harnessing the therapeutic efficacy of oral BV administration.


Determination of catecholamines in human plasma by high-performance liquid chromatography: comparison between a new method with fluorescence detection and an established method with electrochemical detection.
We report a sensitive fluorimetric method, in which catecholamines are concentrated from plasma by liquid-liquid extraction and derivatized with the selective fluorescent agent 1,2-diphenylethyl-enediamine prior to chromatography. Optimal conditions for extraction, derivatization and chromatography were investigated. With alpha-methylnorepinephrine as internal standard, the chromatographic separations are complete within 6 min. Limits of detection are 0.3 pg for norepinephrine and epinephrine and 0.5 pg for dopamine. Coefficients of variation are low (3-7%). Comparison of plasma catecholamine values determined with this method and with an established method with electrochemical detection (n = 135) shows good correlation (r = 0.94-1.00), and regression lines are close to lines of identity.


CRF receptor antagonist attenuates stress-induced noradrenaline release in the medial prefrontal cortex of rats.
Noradrenaline release in rat medial prefrontal cortex (PFC) was measured using a brain microdialysis technique. Immobilization stress increased noradrenaline release to a maximum level of 248.7 +/- 12.8% of the basal release, which was significantly attenuated by preinjection of alpha-helical CRF9-41 (50 micrograms/rat) into the lateral cerebroventricle. Intracerebroventricular injection of CRF also increased noradrenaline release in the medial PFC. These results suggest that immobilization-stress facilitates noradrenaline release in the medial PFC through activation of the CRF system in the brain.


ERbeta Binds N-CoR in the Presence of Estrogens via an LXXLL-like Motif in the N-CoR C-terminus.
Nuclear receptors (NRs) usually bind the corepressors N-CoR and SMRT in the absence of ligand or in the presence of antagonists. Agonist binding leads to corepressor release and recruitment of coactivators. Here, we report that estrogen receptor beta (ERbeta) binds N-CoR and SMRT in the presence of agonists, but not antagonists, in vitro and in vivo. This ligand preference differs from that of ERalpha interactions with corepressors, which are inhibited by estradiol, and resembles that of ERbeta interactions with coactivators. ERbeta /N-CoR interactions involve ERbeta AF-2, which also mediates coactivator recognition. Moreover, ERbeta recognizes a sequence (PLTIRML) in the N-CoR C-terminus that resembles coactivator LXXLL motifs. Inhibition of histone deacetylase activity specifically potentiates ERbeta LBD activity, suggesting that corepressors restrict the activity of AF-2. We conclude that the ER isoforms show completely distinct modes of interaction with a physiologically important corepressor and discuss our results in terms of ER isoform specificity in vivo.


Selective separation of zinc and iron/carbon from blast furnace dust via a hydrometallurgical cooperative leaching method.
Blast furnace dust (BFD) contains ferrous and nonferrous metals and carbon, and is usually categorized as a typical secondary resource and hazardous waste produced by the iron-making process. The thermodynamic calculation and experimental investigation of the selective separation of zinc and iron/carbon from BFD via a NaCl-HCl-H2O system were carried out. Quantitative zinc and iron/carbon separation and recovery was achieved via a zinc-chloride (ZnCli2-i (i = 0, 1, 2, 3, 4)) cooperative leaching methodology using 3 mol/L NaCl at pH of 4, an L/S ratio of 4:1, a leaching temperature of 70 °C, a HCl concentration of 0.25 mol/L, and a leaching time of 2 h. Moreover, the total Cl- concentration used in for leaching was 2.09 mol (as concluded by the dosages of HCl and NaCl, and material). Results demonstrate that zinc was effectively leached from BFD, while the leaching of iron was hindered, in the acidic region. 93.2% of the zinc was extracted into a leaching solution containing 19.8 g/L zinc under the optimal conditions, and the ratio of the leaching agent to BFD was 300 mL to 75 g. The XRD and SEM-EDS analysis results of the residue reveal that ZnO and ZnS were leached, and the zinc-iron spinel (ZnFe2O4) was not leached and was present together with the iron oxide in the leaching residue. Iron and carbon in the leaching residue was enriched from 49.4% to 60.38%, and the iron and carbon were recovered as secondary resources that can be reused in the iron-making system.


Determination of urinary hippuric acid by micellar electrokinetic capillary chromatography.
We propose a method for the simultaneous determination of hippuric acid (HA) and creatinine based on capillary micellar electrokinetic chromatography. Experimental conditions were 20 mM sodium phosphate, pH 7.20, 25 mM sodium dodecyl sulfate, 5% (v/v) acetonitrile. Electropherograms evidenced HA and creatinine peaks in less than 12 min. The method showed good linearity for both analytes and satisfactory within-day precision. The present method, which is accurate, sensitive, rapid and simple, may be applied to single-spot urine samples.


A strategy to prevent future epidemics similar to the 2019-nCoV outbreak.
A novel bat-origin coronavirus emerged in Wuhan, China in December 2019 and continues to spread across China and the world. At the time of writing, a massive global response has been implemented to control the disease as it spreads from person to person. Yet the high-risk human-wildlife interactions and interfaces that led to the emergence of SARS-CoV and of 2019-nCoV continue to exist in emerging disease hotspots globally. To prevent the next epidemic and pandemic related to these interfaces, we call for research and investment in three areas: 1) surveillance among wildlife to identify the high-risk pathogens they carry; 2) surveillance among people who have contact with wildlife to identify early spillover events; and 3) improvement of market biosecurity regarding the wildlife trade. As the emergence of a novel virus anywhere can impact the furthest reaches of our connected world, international collaboration among scientists is essential to address these risks and prevent the next pandemic.


Inhibition of acute platelet thrombosis formation in stenosed canine coronary arteries by specific serotonin 5HT2 receptor antagonist ritanserin.
The aim of the study was to test the hypothesis that platelet serotonin 5HT2 receptors are important in the genesis of thrombosis in stenosed coronary arteries. The specific serotonin 5HT2 receptor antagonist, ritanserin was used as a pharmacological tool to examine the effect of removal of the participation of the 5HT2 receptors on thrombus growth, in a paired statistical design. The study involved 10 open chest anaesthetised dogs, with constrictors of critical diameter applied to the left circumflex coronary artery. Blood flow was monitored in the left circumflex coronary arteries, distal to the critical stenosis. Flow reductions occurred that have previously been shown to be caused by the accumulation of platelet thrombi. By embolising the thrombi, the process could be monitored cyclically (cyclic flow reductions). The specific serotonin 5HT2 receptor antagonist, ritanserin, abolished cyclic flow reductions at a dose of 0.5 mg.kg-1. There was no effect on blood pressure or heart rate on administration of ritanserin at any dose. The serotonin blockade by ritanserin also prevented the reestablishment of cyclic flow reductions by adrenaline infusion (0.4 micrograms.kg-1.min-1), but required ritanserin doses up to 1.5 mg.kg-1. Ex vivo aggregation of platelets was reduced in blood taken from the dogs after ritanserin administration. These results constitute further evidence of the possible importance of serotonin as a mediator of platelet thrombosis in stenosed coronary arteries.


Prognostic value of preoperative C-reactive protein to albumin ratio in patients with thymic epithelial tumors: a retrospective study.
The C-reactive protein to albumin ratio (CAR) is associated with poor prognosis in various cancers. However, its value in thymic epithelial tumors remains to be elucidated, we aimed to evaluate the prognostic significance of preoperative CAR in patients with surgically resected thymic epithelial tumors (TETs). We retrospectively collected data from 125 patients with TETs who underwent thymoma resection at our center. The best cutoff values for the continuous variable, CAR, were obtained using X-tile software. Univariate and multivariate Cox regression analyses were used to evaluate CAR as an independent predictor of overall survival (OS) and recurrence-free survival (RFS). Kaplan-Meier analysis and log-rank tests were used to present risk stratification of patients based on CAR and the Glasgow-prognostic-score (GPS). The prognostic effect of CAR was assessed using a receiver operating characteristic curve. Patients were categorized into high (≥ 0.17) and low (< 0.17) CAR groups according to the optimal cutoff value of 0.17. Univariate and multivariate analyses showed that CAR was an independent predictor of prognosis. World health organization stage, CAR level, GPS score, and drinking history were important independent prognostic factors for OS (p < 0.05). T stage, CAR level, and drinking history were important independent prognostic factors for RFS (p < 0.05). The area under the curve value of CAR to predict prognosis was 0.734 for OS and 0.680 for RFS. Elevated preoperative CAR was independently associated with poor OS and RFS after thymectomy. Therefore, CAR may be a valuable biomarker for the postoperative prognosis of TETs.


Identification of the novel HLA-B*56:01:18 allele using next-generation sequencing.
The HLA-B*56:01:18 allele differs from HLA-B*56:01:01:01 allele by a single nucleotide in codon 326.3.


[Unequal crossing over is the principal pathway of homologous recombination in tandem duplications of Escherichia coli].
Homologous recombination between direct DNA repeats in tandem duplications usually leads to their dissociation. An even number of crossovers between two copies of a duplication should lead to the formation of diploid segregants, i.e., to the preservation of the duplication. However, in studies of the genotype of diploid segregants in heterozygous tandem duplications of Escherichia coli, it was shown that they arise by unequal exchanges between sister chromosomes rather than by intrachromosomal exchanges. Generally, these exchanges lead to the establishment of the homozygous state of (heterozygous) duplications. Since the available data suggest that the exchange between sister chromosomes may be coupled with DNA replication, it is supposed that unequal exchanges between direct DNA repeats occur in the process of DNA replication.


The efficacy of cefazolin plus macrolide (erythromycin or clarithromycin) versus cefazolin alone in neonatal morbidity and placental inflammation for women with preterm premature rupture of membranes.
Although the use of broad-spectrum antibiotics in women with preterm premature rupture of membranes (PPROM) is recommended to prolong pregnancy and decrease short-term neonatal complications, the optimal regimen remains undetermined. The objective of this study was to compare the efficacy of cefazolin plus macrolide (erythromycin or clarithromycin) versus cefazolin alone in reducing neonatal morbidity and placental inflammation for women with PPROM. This prospective study included singleton pregnancies with PPROM (23-33 weeks gestation). The primary outcome was neonatal composite morbidity and the secondary outcomes were the incidence of abnormal brain sonography and infant neurological outcome at one year of age. The presence and the stage of acute histological chorioamnionitis and funisitis were also reviewed blinded to all clinical information. 102 women were randomly assigned to cefazolin (n = 35), cefazolin plus erythromycin (n = 31), or cefazolin plus clarithromycin (n = 36). The neonatal composite morbidity, the incidence of abnormal brain sonography, and infant neurological outcome at one year of age were similar between the comparison treatments (combination of cefazolin plus erythromycin or clarithromycin) and cefazolin. However, the presence and stage of histological funisitis showed significant difference between cefazolin plus clarithromycin versus cefazolin alone (p = 0.023). This study is the first clinical trial of the use of cefazolin with either clarithromycin or erythromycin compared to cefazolin alone in the management of PPROM in which the primary and secondary analyses showed no difference among the three antibiotic regimens. The only noted difference was from a lesser degree of histological funisitis associated with clarithromycin exposure. Our data suggests that clarithromycin may be an alternative worth considering with potentially beneficial effects compared to erythromycin in PPROM.


Is it time to celebrate a century of blood pressure management?
Even after a century of hypertension management, several trivial issues have remained unsolved and still result in inadequate control of blood pressure in many patients. We have performed a critical literature review on topics pertaining the assessment and management of hypertension, focusing on the gaps in knowledge and pitfalls of hypertension management in clinical practice. Physicians are often not capable of adequately measuring blood pressure themselves; the office blood pressure recordings are not sufficiently reliable and the timing of the start of the treatment is often not satisfactory. However, much progress has been made in the past century. Research in this field not only expanded the potential of blood pressure measurements for risk stratification and risk management but also identified many erroneous assumptions and technical mistakes. An important step forward may be the development of affordable 'high tech' blood pressure measuring devices that continuously monitor the intra-arterial pressure in a noninvasive way to eliminate human factors and equipment errors. Technological advances will also provide adequate data storage and software allowing management on an international scale. Hypertension management can almost be celebrated; however, the pressure is still on!


Superoxide modification and inactivation of a neuronal receptor-like complex.
Excessive superoxide (O(-)(2)) formation is toxic to cells and organisms. O(-)(2) reacts with either iron-sulfur centers or cysteines (Cys) of cytoplasmic proteins. Reactions with membrane proteins, however, have not been fully characterized. In the present studies, the reaction of O(-)(2) with a protein complex that has glutamate/N-methyl-D-aspartate (NMDA) receptor characteristics and with one of the subunits of this complex was examined. Exposure of the complex purified from neuronal membranes and the recombinant glutamate-binding protein (GBP) subunit of this complex to the O(-)(2)-generating system of xanthine (X) plus xanthine oxidase (XO) caused strong inhibition of L-[3H]glutamate binding. Inhibition of glutamate binding to the complex and GBP by O(-)(2) was greater than that produced by H(2)O(2), another product of the X plus XO reaction. Mutation of two cysteine (Cys) residues in recombinant GBP (Cys(190,191)) eliminated the effect of O(-)(2) on L-[3H]glutamate binding. Both S-thiolation reaction of GBP in synaptic membranes with [35S]cystine and reaction of Cys residues in GBP with [3H]NEM were significantly decreased after exposure of membranes to O(-)(2). Inhibition of cysteylation of membrane GBP by O(-)(2) was still observed after iron chelation by desferrioxamine, albeit diminished, and was not altered by the presence of catalase. Overall, the results indicated that GBP exposure to O(-)(2) modified Cys residues in this protein. The modification was not characterized but it was probably that of disulfide formation.


Different manifestations of the pathogenity of some strains of Fusarium oxysporum f. sp. pisi.
Different cultivation and morphology characteristics were found in 10 monospore isolates of Fusarium oxysporum f. sp. pisi, obtained from yellowing and wilting plants of pea (Pisum sativum L.). The isolates of the fungus were obtained from distant geographical regions of Czechoslovakia and from various cultivars and hybrids of pea. After inoculation of roots, followed by constant conditions of incubation of the Meteor and Jupiter cultivars having their origin at the Plant-breeding Station at Luzany u Prestic, the isolates caused various symptoms of disease, each isolate showed a different degree of pathogenity. The variability of the pathogenity of the isolates depended on the host. Its manifestation, in turn, depended on the dynamics of growth and development of the pathogen as well as the host. The following symptoms could be observed during the pathogenesis: the dying of cotyledons after the contact of the main root with the inoculum, the dying of young plants (the plants usually forming two stems), wilting of young plants, yellowing of bottom leaves and wilting beginning from the bottom leaves, stunted growth, and plant deformation. The symptoms of disease are related to the changes in vascular system.


Correction to: Stationary and Nonstationary Ion and Water Flux Interactions in Kidney Proximal Tubule: Mathematical Analysis of Isosmotic Transport by a Minimalistic Model.
The chapter 'Stationary and Nonstationary Ion and Water Flux Interactions in Kidney Proximal Tubule: Mathematical Analysis of Isosmotic Transport by a Minimalistic Model' has now been made available open access under a CC BY 4.0 license.


Effect of human interferon preparations on lymphoblastogenesis in Down's syndrome.
The best characterised properties of human interferon, its antiviral (AV) and cell multiplication inhibitory (CMI) activities, are controlled, in an unexplained manner, by genes on chromosomes 21 (refs 1-4; 14-16). Human and animal interferons have various immunosuppressive effects, among them the inhibition in vitro of DNA synthesis in activated lymphocytes. Using mitogen- and antigen-stimulated lymphocytes from normal subjects (disomic 21) and others with Down's syndrome (trisomic 21), we have found that DNA synthesis is inhibited to a greater degree in the latter by both fibroblastoid and leukocyte interferons. We suggest that this property is also regulated by genes on chromosome 21.


A novel fluorinated derivative of diethylstilbestrol.
The synthesis of the diethylstilbestrol (DES) derivative with fluorine atoms present in the positions ortho to the hydroxyl in each ring is described. In vitro studies in a system containing horse radish peroxidase/H2O2 demonstrate extensive oxidation of tetrafluorodiethylstilbestrol to the corresponding dienestrol derivative. Tetrafluorodiethylstilbestrol and DES had comparable in vivo uterotropic activities at a dose of 100 microgram/kg. Competitive binding experiments demonstrated 20-25 fold reduced interaction with the mouse uterine estrogen receptor. This compound may be useful as an experimental estrogen in distinguishing between the biological and toxic effects of DES.


Caspase-8 Variant G Regulates Rheumatoid Arthritis Fibroblast-Like Synoviocyte Aggressive Behavior.
Fibroblast-like synoviocytes (FLS) play a pivotal role in rheumatoid arthritis (RA) by contributing to synovial inflammation and progressive joint damage. An imprinted epigenetic state is associated with the FLS aggressive phenotype. We identified CASP8 (encoding for caspase-8) as a differentially marked gene and evaluated its pathogenic role in RA FLSs. RA FLS lines were obtained from synovial tissues at arthroplasty and used at passage 5-8. Caspase-8 was silenced using small interfering RNA, and its effect was determined in cell adhesion, migration and invasion assays. Quantitative reverse transcription PCR and western blot were used to assess gene and protein expression, respectively. A caspase-8 selective inhibitor was used determine the role of enzymatic activity on FLS migration and invasion. Caspase-8 isoform transcripts and epigenetic marks in FLSs were analyzed in FLS public databases. Crystal structures of caspase-8B and G were determined. Caspase-8 deficiency in RA FLSs reduced cell adhesion, migration, and invasion independent of its catalytic activity. Epigenetic and transcriptomic analyses of RA FLSs revealed that a specific caspase-8 isoform, variant G, is the dominant isoform expressed (~80% of total caspase-8) and induced by PDGF. The crystal structures of caspase-8 variant G and B were identical except for a unique unstructured 59 amino acid N-terminal domain in variant G. Selective knockdown of caspase-8G was solely responsible for the effects of caspase-8 on calpain activity and cell invasion in FLS. Blocking caspase-8 variant G could decrease cell invasion in diseases like RA without the potential deleterious effects of nonspecific caspase-8 inhibition.


Detection of polyol accumulation in a new ovarian carcinoma cell line, CABA I: a(1)H NMR study.
Ovarian carcinomas represent a major form of gynaecological malignancies, whose treatment consists mainly of surgery and chemotherapy. Besides the difficulty of prognosis, therapy of ovarian carcinomas has reached scarce improvement, as a consequence of lack of efficacy and development of drug-resistance. The need of different biochemical and functional parameters has grown, in order to obtain a larger view on processes of biological and clinical significance. In this paper we report novel metabolic features detected in a series of different human ovary carcinoma lines, by (1)H NMR spectroscopy of intact cells and their extracts. Most importantly, a new ovarian adenocarcinoma line CABA I, showed strong signals in the spectral region between 3.5 and 4.0 p.p.m., assigned for the first time to the polyol sorbitol (39+/-11 nmol/10(6) cells). (13)C NMR analyses of these cells incubated with [1-(13)C]-D-glucose demonstrated labelled-sorbitol formation. The other ovarian carcinoma cell lines (OVCAR-3, IGROV 1, SK-OV-3 and OVCA432), showed, in the same spectral region, intense resonances from other metabolites: glutathione (up to 30 nmol/10(6) cells) and myo-inositol (up to 50 nmol/10(6) cells). Biochemical and biological functions are suggested for these compounds in human ovarian carcinoma cells, especially in relation to their possible role in cell detoxification mechanisms during tumour progression.


A balloon protection sheath to prevent peripheral embolization during aortoiliac endovascular procedures.
Peripheral embolization is a known complication occurring during aortoiliac recanalization. In particular, acute, fresh thrombotic occlusions, with a history shorter than 6 months, have a high risk for peripheral embolization. We describe a balloon occlusion sheath used to manage embolic material while preventing peripheral embolization.


When to seek health care: a duration analysis for malaria patients in Nepal.
We find that the log-normal distribution of care-seeking time - the number of days from the onset of symptoms of malaria to when a patient seeks treatment from a provider - best described the treatment-seeking behavior of malaria patients in rural areas of two districts of Nepal. The care-seeking rate, or the probability of seeking care, was low on the first day of the symptoms; it increased sharply over the first five days and then gradually declined. Since at the time of the research there was a system of malaria workers taking monthly surveillance rounds of each house to detect and treat malaria cases, patients, instead of traveling to a provider for care, generally waited for malaria workers to arrive at home when the wait for malaria workers was short. But, the probability of seeking care on any day rose if the wait was longer. Women generally tended to wait longer for the malaria workers in order to receive treatment at home. Patient's age, household size, education, and the type of malaria species infecting the patient had no significant effect on care-seeking rate. Given an assumption that a wait of 100 days for a malaria worker would effectively represent total absence of surveillance program, the estimated model predicted higher care-seeking rates under no surveillance program than under the monthly surveillance program.


Closed Shell Iron(IV) Oxo Complex with an Fe-O Triple Bond: Computational Design, Synthesis, and Reactivity.
Iron(IV)-oxo intermediates in nature contain two unpaired electrons in the Fe-O antibonding orbitals, which are thought to contribute to their high reactivity. To challenge this hypothesis, we designed and synthesized closed-shell singlet iron(IV) oxo complex [(quinisox)Fe(O)]+ (1+ ; quinisox-H=(N-(2-(2-isoxazoline-3-yl)phenyl)quinoline-8-carboxamide). We identified the quinisox ligand by DFT computational screening out of over 450 candidates. After the ligand synthesis, we detected 1+ in the gas phase and confirmed its spin state by visible and infrared photodissociation spectroscopy (IRPD). The Fe-O stretching frequency in 1+ is 960.5 cm-1 , consistent with an Fe-O triple bond, which was also confirmed by multireference calculations. The unprecedented bond strength is accompanied by high gas-phase reactivity of 1+ in oxygen atom transfer (OAT) and in proton-coupled electron transfer reactions. This challenges the current view of the spin-state driven reactivity of the Fe-O complexes.


Changes in level of precision of common ligand assays during a seven-year interval.
Nine commonly performed radioligand analyses were studied for periods of four to seven years in terms of change in level of precision. Consistent improvement in precision is generally observed, except in specific instances when pool characteristics are altered to study sensitivity and crossreactivity.


Esophago-tracheobronchial fistula following treatment of anlotinib in advanced squamous cell lung cancer: Two case reports.
Anlotinib, a novel orally administered multitargeted tyrosine kinase inhibitor, inhibiting tumor angiogenesis and growth, significantly prolonged overall survival, and progression-free survival with a manageable safety profile as a third-line therapy among refractory advanced nonsmall cell lung cancer (NSCLC) patients in ALTER 0303 trail (NCT02388919). Two squamous cell lung cancer patients with mediastinal metastasis undergoing the treatment of anlotinib developed clinical symptom of cough, which was worse upon ingestion. On the basis of patients' clinical symptoms and radiographic findings, they were diagnosed with acquired esophago-tracheobronchial fistula. We treated them with fully covered self-expandable metallic stents. The clinical symptom of cough was immediately relieved after palliative treatment. Both patients elected to discontinue anlotinib treatment. These 2 cases imply that a close follow-up schedule for esophago-tracheobronchial fistula should be established when squamous cell lung cancer patients with mediastinal metastasis are undergoing anlotinib therapy. Early detection and adequate treatment are essential for patient symptom relief and survival.


Comparison of incidence/risk of venous thromboembolism (VTE) among selected clinical and hereditary risk markers: a community-based cohort study.
Little information is available from community-based long-term VTE cohort studies to compare the absolute thrombosis risk of established clinical and genetic risk factors. The occurrence of venous thromboembolism (VTE) was observed during a 10-year observation period in the BAvarian ThromboEmbolic Risk (BATER) study, a cohort study of 4337 women (age 18-55 years). We collected data on demographics, reproductive life, lifestyle, conditions/diseases, and particularly potential risk factors for VTE with a self-administered questionnaire. The objective was to present incidence rates of VTE and to show relative risk estimated associated with different clinical and genetic risk factors. 34 new, by diagnostic means confirmed VTE events occurred during the observation time of 32,656 women-years (WY). The overall incidence of VTE was 10.4 per 10(4) WY. The incidence rates varied markedly among different risk cohorts. The highest incidence was observed in women with previous history of VTE, followed by family history of VTE. None of the measured "genetically-related risk markers" (antithrombin, protein C, FVL, prothrombin mutation, or MTHFR) showed a significant VTE risk. Most of the discussed VTE risk factors showed no significant association with the occurrence of new VTEs due to smallness of numbers. Only first-degree family history of VTE and own history of a previous VTE event depicted a significant association with future VTE. Clinical information seems to be more important to determine future VTE risk than genetically related laboratory tests.


Inhibition of cell cycle-dependent hyphal and biofilm formation by a novel cytochalasin 19,20‑epoxycytochalasin Q in Candida albicans.
Biofilm-mediated drug resistance is a key virulence factor of pathogenic microbes that cause a serious global health threat especially in immunocompromised individuals. Here, we investigated the antihyphal and antibiofilm activity of 19,20‑epoxycytochalasin Q (ECQ), a cytochalasin actin inhibitor isolated from medicinal mushroom Xylaria sp. BCC1067 against Candida albicans. Remarkably, 256 µg/ml of ECQ inhibited over 95% of C. albicans hyphal formation after 24 h-treatment. Combined ECQ and lipid-based biosurfactant effectively enhanced the antihyphal activity, lowering required ECQ concentrations. Hyphal fragmentation and reduction of biofilm biomass, shown by SEM and AFM visualization of ECQ-treated biofilms, were well corelated to the reduced metabolic activities of young and 24 h-preformed C. albicans biofilms. Induced intracellular accumulation of reactive oxygen species (ROS) also occurred in accompany with the leakage of shrunken cell membrane and defective cell wall at increasing ECQ concentrations. Transcriptomic analyses via RNA-sequencing revealed a massive change (> 1300 genes) in various biological pathways, following ECQ-treatment. Coordinated expression of genes, associated with cellular response to drugs, filamentous growth, cell adhesion, biofilm formation, cytoskeleton organization, cell division cycle, lipid and cell wall metabolisms was confirmed via qRT-PCR. Protein-protein association tool identified coupled expression between key regulators of cell division cyclin-dependent kinases (Cdc19/28) and a gamma-tubulin (Tub4). They coordinated ECQ-dependent hyphal specific gene targets of Ume6 and Tec1 during different phases of cell division. Thus, we first highlight the antihyphal and antibiofilm property of the novel antifungal agent ECQ against one of the most important life-threatening fungal pathogens by providing its key mechanistic detail in biofilm-related fungal infection.


Homocysteine measurement by Vitros Microtip homocysteine assay.
Elevated circulating total homocysteine is an independent vascular risk factor. Enzymatic homocysteine measurements represent an alternative to HPLC- or immunochemistry-based assays, suitable for automation. Here, we report on analytical performance of a commercial cystathionine beta-synthase-based assay, for use on Vitros automated analyzers. Linear range, limit of detection and analytical sensitivity were inferred from duplicate measurements of homocystine standard solutions (1-65 micromol/L). Imprecision was assessed using commercial controls according to NCCLS EP5-A2 and accuracy using NIST-SRM1955 reference material. Agreement with a clinically validated HPLC method was examined on 207 patient samples. The enzymatic assay was linear from 1 to 90 micromol/L homocysteine. Total (within-day) imprecision ranged from 4.5 (3.9)% to 2.8 (1.6)% at homocysteine 9.7-43.2 micromol/L. Accuracy was acceptable at 8.9 and 17.7 micromol/L homocysteine, with +6.4% and -1.2% bias, respectively, but showed substantial negative bias (-20.1%) at 4.0 micromol/L. High triglycerides (19.8 micromol/L) negatively interfered. The enzymatic method was slightly less sensitive than the HPLC method (limit of detection 0.7 and 0.2 micromol/L, respectively) but correlated well with the latter (r2=0.9997, slope=1.04, intercept=-0.66 micromol/L) and was more precise (p<0.05). The Vitros homocysteine assay met the CLIA Desirable Analytical Quality Specifications at homocysteine > or = 9 micromol/L. Its analytical performance and suitability for automation make the Vitros assay an analytically acceptable alternative to HPLC-based methods.


Doppler sonography for predicting fetal anemia caused by massive fetomaternal hemorrhage.
Fetomaternal hemorrhage (FMH) can cause severe anemia in the fetus. Untreated, this may cause hydrops or even fetal death. However, correct diagnosis of FMH followed by blood transfusion can prevent these life-threatening consequences. We describe two cases in which fetal anemia was suspected because of maternal reporting of decreased or absent fetal movements, the detection of a sinusoidal heart rate pattern and increased blood flow velocities of the middle cerebral artery and umbilical vein. Together with the Betke-Kleihauer test showing fetal cells in the maternal circulation, this led to the correct diagnosis of severe fetal anemia caused by FMH. A Cesarean section was performed within a few hours. Both neonates were severely anemic and received immediate blood transfusions. They are currently alive and well.


Verified spider bites in Oregon (USA) with the intent to assess hobo spider venom toxicity.
This study compiled 33 verified spider bites from the state of Oregon (USA). The initial goal was to amass a series of bites by the hobo spider to assess whether it possesses toxic venom, a supposition which is currently in a contested state. None of the 33 bites from several spider species developed significant medical symptoms nor did dermonecrosis occur. The most common biters were the yellow sac spider, Cheiracanthium mildei (N = 10) and orb-weavers of the genus Araneus (N = 6). There were 10 bites from three genera of funnel web spiders of the family Agelenidae including one hobo spider bite and one from the congeneric giant house spider which is readily confused as a hobo spider. The hobo spider bite resulted in pain, redness, twitching in the calf muscle and resolved in 12 h. Also generated from this study were possibly the first records of bites from spiders of the genera Callobius (Amaurobiidae) and Antrodiaetus (Antrodiaetidae), both with minor manifestations.


Botulinum toxin-A injections for spastic toe clawing.
Spastic toe clawing describes extension at the metatarsophalangeal joints of the feet, flexion at the proximal interphalangeal joints and flexion at the distal interphalangeal joints that results from upper motor neuron lesions, such as stroke, intracranial hemorrhage, cervical myelopathy and brain tumors. Even though toe clawing is often asymptomatic, it can be painful. Previous studies have described the efficacy of injections of botulinum toxin type-A (BTX-A) to the long flexors of the toes, but this is often unsatisfactory as high dosages (up to 175 units) have been required, and patients often report significant residual toe clawing. We performed an open label, prospective study to assess the efficacy of BTX-A injections, targeting the long and short flexors of the toes, performed with electrical (motor point) stimulation under electromyographic guidance. Outcome measures, which included timed walking over 20m, objective assessment of toe clawing (modified Ashworth scale and a visual analog scale rating) and patient assessment of functional disability, were assessed before injections and at six-weeks' follow-up. Seven patients (five male and two female) of mean age 51 (range 38-70) were recruited. Four had spasticity from underlying intracranial hemorrhage, the remaining three from cerebral infarct, astrocytoma and post-traumatic cervical myelopathy. The total dose of BTX-A injected for toe clawing ranged from 40 to 90 units. Improvements were observed in all outcome measures except timed walking. Injecting BTX-A to the long and short flexors of the toes, with electrical stimulation under electromyographic guidance, is well tolerated and efficacious in the treatment of toe clawing from spasticity, allowing for lower dosages to be used.


A sample-to-answer COVID-19 diagnostic device based on immiscible filtration and CRISPR-Cas12a-assisted detection.
In response to the ongoing coronavirus disease 2019 (COVID-19) pandemic and disparities of vaccination coverage in low-and middle-income countries, it is vital to adopt a widespread testing and screening programme, combined with contact tracing, to monitor and effectively control the infection dispersion in areas where medical resources are limited. This work presents a lab-on-a-chip device, namely 'IFAST-LAMP-CRISPR', as an affordable, rapid and high-precision molecular diagnostic means for detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). The herein proposed 'sample-to-answer' platform integrates RNA extraction, amplification and molecular detection with lateral flow readout in one device. The microscale dimensions of the device containing immiscible liquids, coupled with the use of silica paramagnetic beads and guanidine hydrochloride, streamline sample preparation (including RNA extraction, concentration and purification) in 15 min with minimal hands-on steps. The pre-amplification in combination with CRISPR-Cas12a detection assays targeting the nucleoprotein (N) gene achieved visual identification of ≥ 470 copies mL-1 genomic SARS-CoV-2 samples in 45 min. On-chip assays showed the ability to isolate and detect SARS-CoV-2 RNA from 100 genome copies mL-1 of replication-deficient viral particles in 1 h. This simple, affordable and integrated platform demonstrated a visual, faster, and yet specificity- and sensitivity-comparable alternative to the costly gold-standard reverse transcription-polymerase chain reaction (RT-PCR) assay, requiring only a simple heating source. Initial testing illustrates the platform viability both on nasopharyngeal swab and saliva samples collected using the easily accessible Swan-brand cigarette filter, providing a complete workflow for COVID-19 diagnostics in low-resource settings.


Fetal outcomes after maternal exposure to oral antifungal agents during pregnancy: A systematic review and meta-analysis.
To assess the risk of adverse fetal outcomes after exposure to oral antifungal agents during pregnancy. PubMed, Embase, and Cochrane Central Register of Controlled Trials (CENTRAL) were searched up to October 2018. Cohort studies and case-control studies investigating fetal outcomes following maternal exposure to oral antifungal agents. Two reviewers independently assessed studies for inclusion, assessed risk of bias, and extracted data. Pooled estimates were calculated for the frequency of adverse fetal outcomes. Overall, eight cohort studies and one case-control study were included. The oral antifungal agents used during pregnancy were fluconazole and itraconazole. The data indicated that oral fluconazole exposure during pregnancy might slightly increase the risk of congenital heart defects and limb defects relative to the general population; oral itraconazole during pregnancy might increase the risk of eye defects. No difference was found between oral fluconazole/itraconazole exposure and non-exposure in the risk of other birth defects, spontaneous abortion, or stillbirth. Oral fluconazole or itraconazole may not increase the risk of birth defects. Nonetheless, the risk of congenital heart defects and limb defects after fluconazole exposure and eye defects after itraconazole exposure should be cautiously investigated.


Real-time analysis ambulatory electrocardiography--clinical evaluation of cardiac arrhythmias by the Aegis system.
To evaluate the technical reliability, clinical applicability, and arrhythmia accuracy of one manufacturer's real-time analysis ambulatory ECG instrument (Aegis Medical Systems), 164 patients were simultaneously examined through a "Y" cable by both a real-time analyzer and a conventional Holter recorder. Technical failure was similar for both recorders (2% in each), and the real-time analyzer was applicable to all patients encountered. Using a randomly selected hand-counted database of 799 hours as the standard of truth, accuracy of the real-time analyzer for hourly mean heart rate, isolated ventricular ectopic beats, ventricular couplets, ventricular tachycardia, isolated supraventricular ectopic beats and supraventricular tachycardia was determined. Mean heart rate showed 96% agreement and a high correlation (r = .986) to hand-counted values. Real-time analysis overall sensitivity, positive predictive accuracy, and false positive rate for the Aegis Medical System were (a) for isolated ventricular ectopic beats--92%, 92%, and 8%, (b) for ventricular couplets--80%, 97%, and 3%, (c) ventricular tachycardia--81%, 92%, and 8%, (d) supraventricular ectopic beats--81%, 82%, and 18%, and (e) for supraventricular tachycardia--75%, 89%, and 11%. Arrhythmia analysis was valid for mean heart rate and ventricular arrhythmias, but showed lower sensitivities in detection of low prevalence (1-60 b/Hr) ventricular couplets (72%) and ventricular tachycardia (43%). Significant inaccuracies for some densities of supraventricular arrhythmia were also found. This study found the real-time analysis ambulatory ECG similar to conventional Holter recording for technical reliability and patient applicability. Given the limitations of currently available real-time analysis ambulatory ECG systems, it is a promising technology.


Ultraviolet sensitivity ofBacillus subtilis citD mutants.
Cells ofBacillus subtilis 168 with deletions in thecitD locus were found to be sensitive to irradiation with ultraviolet light and to mitomycin C but were able to repair DNA damage induced by methyl methanesulfonate. The recombination abilities of these cells, as determined by transformation and PBS1-mediated transduction experiments, were unaffected by the deletion. These phenotypic characteristics do not result from a metabolic imbalance caused by the deficiency of a functional α-ketoglutarate dehydrogenase complex, but most likely are a consequence of a genetic locus involved in ultraviolet repair, which is deleted together with thecitK gene when the deletion is formed.


Dental Age Estimation-Root Pulp Visibility (RPV) patterns: A reliable Mandibular Maturity Marker at the 18 year threshold.
The purpose of this paper was to explore the potential application of Root Pulp Visibility (RPV) to determine whether or not a subject of unknown date of birth is under or over the 18year threshold. Two thousand Dental Panoramic Radiographs comprising equal numbers of females and males evenly distributed between the ages of 16 and 26 years were examined. All third molars were scored using the Anglo-Canadian 8 Stage Tooth Development system [1]. The Lower Left Third Molar was censored to avoid redundant data being included in the data array for the Age at Attainment of Stage H, the final stage of tooth development (Roberts et al., 2015 [2]). A filter cascade was applied to the data to select the data array for 1. Teeth in the final stage of development, 2. Female or male gender, 3. The definitive stage of RPV: RPV-A, RPV-B, RPV-C, or RPV-D. The summary data for each of these stages was derived and the minimum value for each was used to determine the relationship of the minimum value of A, B, C or D to the 18 Year threshold. It was found that for both females and males the minimum value for RPV-A and RPV-B indicated that subjects displaying this could be under 18 years old. For RPV-C and RPV-D, the minimum values show unequivocally that for both females and males the subject was over 18 years old. RPV has an important role in unambiguously determining whether a subject is below or above the 18year threshold.


Reproducibility of brain activation during auditory verbal hallucinations.
Previous studies investigated fMRI-guided repetitive Transcranial Magnetic Stimulation (rTMS) as an alternative treatment for auditory verbal hallucinations (AVH). This tailor-made treatment focuses at directing the rTMS coil to the location where hallucinatory activation is maximal, as identified with fMRI scans of individual patients. For the effective use of such treatment it is important to determine whether brain activation during AVH can be reliably detected using fMRI. Thirty-three psychotic patients indicated the presence of AVH during two subsequent scans. Reproducibility was measured by calculating 1) the distance between local maxima of significantly activated clusters and 2) percentage overlap of activation patterns over the two scans. These measurements were obtained both in single subjects and on group-level in five regions of interest (ROIs). ROIs consisted of the areas that were most frequently activated during AVH. Scans were considered reproducible if the distance between local maxima was smaller than 2 cm, as rTMS-treatment may target an area of approximately 2-4 cm. The median distance between local maxima was smaller than 2 cm for all ROIs on single-subject level, as well as on group-level. In addition, on single-subject level median percentage overlap varied between 14 and 38% for the different ROIs. On group-level, this was substantially higher with percentages overlap varying between 34 and 98%. Based on these results, AVH-scans may be considered sufficiently reproducible to be suitable for fMRI-guided rTMS treatment.


Characterization of vaginal microflora of healthy, nonpregnant women by chaperonin-60 sequence-based methods.
The purpose of this study was to use a novel method that was based on the application of chaperonin-60 sequencing to describe the vaginal microflora of 16 healthy women. Asymptomatic women consented for vaginal swabs to be collected at the time of a clinical pelvic examination. Total genomic DNA was isolated from the vaginal swabs. Degenerate, universal polymerase chain reaction primers were used to amplify an approximately 555 base pair region of the universal chaperonin-60 gene, which is found in all eubacteria and eukaryotes, from the total genomic DNA and libraries of cloned polymerase chain reaction products were constructed. Library clones were sequenced, and the resulting sequences were assigned to taxonomic groups on the basis of similarity to reference sequence data. Presence of Chlamydophila psittaci sequences in the samples was confirmed by species-specific polymerase chain reaction. Sixteen of the 23 women who were enrolled had normal flora by Nugent's score of <4 and had adequate polymerase chain reaction product for assessment. Vaginal flora libraries were dominated by a variety of sequences with similarity to Lactobacillus spp L. crispatus, L. iners, L. gasseri, L. jensenii, and L. buchneri. Other sequences that were identified included representatives of Gardnerella spp, sequences with similarity to Porphyromonas spp and Megasphaera spp and sequences identical to C psittaci. Culture-independent, chaperonin-60 sequence-based molecular methods can lead to the identification of greater diversity within defined taxa compared with those that are identified by standard culture-based methods and to the identification of novel organisms that were not previously associated with vaginal flora.


Fatal outcome after heart surgery in PMM2-CDG due to a rare homozygous gene variant with double effects.
Variants in Phosphomannomutase 2 (PMM2) lead to PMM2-CDG, the most frequent congenital disorder of glycosylation (CDG). We here describe the disease course of a ten-month old patient who presented with the classical PMM2-CDG symptoms as cerebellar hypoplasia, retinitis pigmentosa, seizures, short stature, hepato- and splenomegaly, anaemia, recurrent vomiting and inverted mamillae. A severe form of tetralogy of Fallot was diagnosed and corrective surgery was performed at the age of 10 months. At the end of the cardiopulmonary bypass, a sudden oedematous reaction of the myocardium accompanied by biventricular pump failure was observed immediately after heparin antagonization with protamine sulfate. The patient died seven days after surgery, since myocardial function did not recover on ECMO support. We here describe the first patient carrying the homozygous variant g.18313A > T in the PMM2 gene (NG_009209.1) that either can lead to c.394A > T (p.I132F) or even loss of 100 bp due to exon 5 skipping (c.348_447del; p.G117Rfs*4) which is comparable to a null allele. Proliferation and doubling time of the patient's fibroblasts were affected. In addition, we show that the induction of cellular stress by elevating the cell culture temperature to 40 °C led to a decrease of the patients' PMM2 transcript as well as PMM2 protein levels and subsequently to a significant loss of residual activity. We assume that metabolic stressful processes occurring after cardiac surgery led to the drop of the patient's PMM activity below a life-sustaining niveau which paved the way for the fatal outcome.


Transposon insertion resulted in the silencing of Wx-B1n in Chinese wheat landraces.
A novel Wx-B1 allele was characterized; a transposon insertion resulted in the loss of its function, which is different from the previously reported gene silencing mechanisms at the Wx-B1 locus. The waxy protein composition of 53 Chinese wheat landraces was analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis; of these, 10 did not show the expression of Wx-A1 (four accession) or Wx-B1 (six accessions) protein. The results of molecular marker detection revealed that the Wx-B1 allele (Wx-B1n) showed normal expression, inconsistent with the findings of SDS-PAGE for the Xiaobaipi accession. Further cloning of the 9160-bp region covering the Wx-B1 coding region and 3'-downstream region revealed that a 2178-bp transposon fragment had been inserted at 2462 bp within the tenth exon of Wx-B1n ORF, leading to the absence of Wx-B1 protein. Sequence analysis indicated that the insertion possessed the structural features of invert repeat and target repeat elements, we deduced that it was a transposon. Further PCR analysis revealed that this fragment had moved, but not copied itself, from 3B chromosome to the current location in Wx-B1n. Therefore, the reason for the inactivation of Wx-B1n was considerably different from those for the inactivation of Wx-B1b, Wx-B1k, and Wx-B1m; to our knowledge, this kind of structural mutation has never been reported in Wx-B1 alleles. This novel allele is interesting, because it was not associated with the deletion of other quality-related genes included in the 67 kb region lost with the common null allele Wx-B1b. The null Wx-B1n might be useful for investigating gene inactivation and expression as well as for enriching the genetic resource pool for the modification of the amylose/amylopectin ratio, thereby improving wheat quality.


DDT and HCH residue load in mother's breast milk: a survey of lactating mother's from remote villages in Agra region.
Breast milk is the natural and optimal food for the infants. In addition to meeting the nutritional needs, breast milk provides numerous immunological, developmental, psychological, ecological and practical advantages. The levels of the DDT and HCH residues in human milk collected from remote rural part of Agra were analysed. Beta and gamma isomers of HCH accounts for the major residue of total HCH excreted in breast milk. Delta-HCH was not detected in the samples. 95% of the samples were found to be contaminated with DDT and its metabolites. DDD was found in 88% of the samples analyzed. The total concentration of DDT and HCH were found lower than the previous studies carried out in India. The study shows the decreasing levels of these organochlorine pesticides from the environment. Total concentration of total DDT is higher than total HCH.


Active digital microfluidic paper chips with inkjet-printed patterned electrodes.
Active, paper-based, microfluidic chips driven by electrowetting are fabricated and demonstrated for reagent transport and mixing. Instead of using the passive capillary force on the pulp to actuate a flow of a liquid, a group of digital drops are transported along programmed trajectories above the electrodes printed on low-cost paper, which should allow point-of-care production and diagnostic activities in the future.


Factors associated with nursing students' willingness to care for older adults in Korea and the United States.
Elderly health care needs increase and nurses' role for elderlies is vital. It is significant to identify nursing students' intention to care for elderlies. Thus, this study investigated nursing students' willingness to care for elderlies in Korea and the United States. The study was conducted with 437 undergraduate nursing students from Korea and the United States from May 25 to 31, 2018. Participants completed a survey including frequency and quality of contact, anxiety about aging, empathy, attitude toward elderly, and willingness to care. Study findings from the entire group showed that nursing students' willingness to care for the elderly was positively associated with contact quality (β = 0.22, P < 0.001) and empathy (β = 0.12, P = 0.009) but negatively associated with anxiety about aging (β = -0.23, P < 0.001) and attitude toward the elderly (β = -0.14, P = 0.004). Contact quality (β = 0.30, P < 0.001) was positively associated with the willingness to care in Korean students, whereas extended family living type (β = -0.15, P = 0.012) and attitude toward the elderly (β = -0.18, P = 0.005) negatively associated in US students. This study suggested that nursing educators reinforce contact quality and empathy but reduce anxiety about aging and attitude toward elderly to enhance future nurses' care quality.


Targeting of Disordered Proteins by Small Molecules in Neurodegenerative Diseases.
The formation of protein aggregates and inclusions in the brain and spinal cord is a common neuropathological feature of a number of neurodegenerative diseases including Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and many others. These are commonly referred as neurodegenerative proteinopathies or protein-misfolding diseases. The main characteristic of protein aggregates in these disorders is the fact that they are enriched in amyloid fibrils. Since protein aggregation is considered to play a central role for the onset of neurodegenerative proteinopathies, research is ongoing to develop strategies aimed at preventing or removing protein aggregation in the brain of affected patients. Numerous studies have shown that small molecule-based approaches may be potentially the most promising for halting protein aggregation in neurodegenerative diseases. Indeed, several of these compounds have been found to interact with intrinsically disordered proteins and promote their clearing in experimental models. This notwithstanding, at present small molecule inhibitors still awaits achievements for clinical translation. Hopefully, if we determine whether the formation of insoluble inclusions is effectively neurotoxic and find a valid biomarker to assess their protein aggregation-inhibitory activity in the human central nervous system, the use of small molecule inhibitors will be considered as a cure for neurodegenerative protein-misfolding diseases.


Population coding of saccadic eye movements by neurons in the superior colliculus.
The deeper layers of the superior colliculus are involved in the initiation and execution of saccadic (high velocity) eye movements. A large population of coarsely tuned collicular neurons is active before each saccade. The mechanisms by which the signals that precisely control the direction and amplitude of a saccade are extracted from the activity of the population are unknown. It has been assumed that the exact trajectory of a saccade is determined by the activity of the entire population and that information is not extracted from only the most active cells in the population at a subsequent stage of neural processing. The trajectory of a saccade could be based on vector summation of the movement tendencies provided by each member of the population of active neurons or be determined by a weighted average of the vector contributions of each neuron in the active population. Here we present the results of experiments in which a small subset of the active population was reversibly deactivated with lidocaine. These results are consistent with the predictions of the latter population-averaging hypothesis and support the general idea that the direction, amplitude and velocity of saccadic eye movements are based on the responses of the entire population of cells active before a saccadic eye movement.


Immunization with meningococcal polysaccharide-tetanus toxoid conjugate induces polysaccharide-reactive T cells in mice.
T cell clones were generated from mice immunized with a meningococcal group C (alpha2 --> 9-sialic acid) polysaccharide-tetanus toxoid (MCPS-TT) conjugate. Many clones were found to be specific for tetanus toxoid (TT), however, clones reactive with MCPS-TT and polysaccharide (PS) were isolated. Two clones were specific for MCPS and two cross-reacted with Escherichia coli K1-PS (alpha2 --> 8-sialic acid). Both TT and PS reactive clones were CD4+ and CD8-. TT and MCPS-TT-specific T cell clones were major histocompatibility complex (MHC) restricted, however, the PS-reactive clones were not. Both MHC-restricted TT clones and non-restricted PS clones, however, were dependent on contact with antigen presenting cells (APC) for maximal stimulation. The data suggest that multivalent repeating epitopes on PS antigen (Ag) can overcome the need for MHC restricted interactions, but not the requirement for cell-cell contact.


First aid intervention with mothers/caregivers of children affected by the Zika virus in Brazil.
Children with microcephaly associated with the Zika virus are more likely to develop choking and apnea, so it is necessary that mothers of this population receive first aid training to cope with emergency situations that their children may experience. To understand how an educational first aid intervention with mothers/caregivers of Brazilian children with congenital Zika virus syndrome impacts their management of seizures and choking. A qualitative study conducted in a philanthropic institution with ten mothers/caregivers of children with congenital Zika virus syndrome. Data were collected between August and November 2018 through four focus groups with two meetings for intervention/educational workshops in first aid on situations of choking and seizure. After one month, two more meetings were held to evaluate the effectiveness of this action performed at home. The empirical material was submitted to Content Analysis, and discussed in the light of Paulo Freire's Pedagogy of Autonomy. The mothers/caregivers performed ineffective and harmful behaviors to the children in situations of choking and seizure before the intervention. Thus, the intervention empowered these caregivers through an exchange of experiences which began to present discernment, autonomy, resolving capacity and confidence to act in these experienced situations. They also became knowledge disseminators for family members and neighbors. The educational intervention can have satisfactory results in training mothers/caregivers in first aid, highlighting the importance of this initiative to reduce the morbidity and mortality of children who experience choking and seizures.


Unilateral nephrectomy of cattle.
Renal disease is occasionally diagnosed in cattle. Most often, renal disease is bilateral and medical therapy is the treatment of choice. Nephrectomy may be performed in selected cases where a single kidney is primarily involved and medical therapy is unsuccessful. This surgery can be performed under field settings, and these cattle can remain productive members of the herd.


Bacterial Abundance and Activity across Sites within Two Northern Wisconsin Sphagnum Bogs.
Abstract Bacterial abundance, temperature, pH, and dissolved organic carbon (DOC) concentration were compared across surface sites within and between two northern Wisconsin Sphagnum peatlands over the summer seasons in 1995 and 1996. Sites of interest were the Sphagnum mat surface, the water-filled moat (lagg) at the bog margin, and the bog lake littoral zone. Significant differences in both bacterial populations and water chemistry were observed between sites. pH was highest in the lake and lowest in the mat at both bogs; the opposite was true for DOC. Large populations of bacteria were present in surface interstitial water from the mat; abundance in this site was consistently higher than in the moat or lake. Bacterial abundance also increased across sites of increasing DOC concentration and declining pH. Bacterial activities (rates of [3H]leucine incorporation) and growth in dilution cultures (with grazers removed) were also assessed in lake, moat, and mat sites. Results using these measures generally supported the trends observed in abundance, although high rates of [3H]leucine incorporation were recorded in the moat at one of the bogs. Our results indicate that bacterial populations in Sphagnum peatlands are not adversely affected by acidity, and that DOC may be more important than pH in determining bacterial abundance in these environments.


Clinicopathology of graft detachment after Descemet's membrane endothelial keratoplasty.
To investigate pathomechanisms involved in graft detachment after Descemet's membrane endothelial keratoplasty (DMEK) and its clinical implications. In a prospective case series, 30 eyes with Fuchs' endothelial dystrophy underwent DMEK. Intraoperatively obtained recipients' endothelium-Descemet's membranes (EDMs) were investigated histologically and immunohistochemically. The postoperative donor graft status was categorized as attached or detached. Clinical and morphological parameters were analysed between the study groups. The detachment rate was 40% (12/30). There was no significant difference in postoperative visual acuity between the groups, but visual recovery was delayed in eyes with initially detached grafts. Gender, age, preoperative central corneal thickness or best corrected visual acuity did not influence the detachment rate. However, separation and disruption of the anterior banded layer (ABL) were frequently observed in patients with graft detachment, and ABL thickness was identified as a significant predictor for graft detachment. The ABL thickness was 2.5 ± 0.9 μm and 3.5 ± 1.6 μm in patients with attached and detached grafts, respectively. Immunohistologically, a deficiency of fibronectin and cytokeratin was observed within the ABL of patients with detached grafts. In contrast, a complete removal of the EDM with residual stromal collagen fragments was observed in patients with adherent grafts. Incomplete removal of the EDM, with residual ABL fragments on the recipients' corneal stroma, may be a risk factor for graft detachment after DMEK. The separation and disruption of the ABL might be promoted by a deficiency of matrix proteins, stronger biomechanical properties and a firm adherence to the posterior corneal stroma.


Some current issues in breast cancer screening.
Randomized trials of mammography have demonstrated the efficacy of mammographic screening for breast cancer in terms of preventing deaths, but various issues of particular interest remain, including: quantification of overdiagnosis; evaluation of service screening outside the research setting; absolute benefit in terms of number needed to screen per life saved; which types of tumours benefit most from early detection; use of screening data to investigate tumour biology and natural history. This paper describes examples of approaches to the above issues, along with some important results.


The accuracy of midwives' clinical breast examination in detection of breast lumps.
Screen-detected breast cancers are usually diagnosed at earlier stages. Therefore, countries with limited resources are recommended to implement clinical breast examination (CBE) as a screening method in conjunction with mammography. Since there are so many limitations to performance of CBE by surgeons in the health system and CBE by midwives is more feasible, this study was conducted to test the agreement of CBE by midwives and surgeons. One thousand and twenty seven patients with no personal history of breast cancer received breast physical examination by both a midwife and a surgeon and designed forms including patients' general information, cause of referral and abnormal physical findings were completed for each patient. The inter-observer agreement (kappa) for mass detection was 36 % (95% CI= 31% to 41%), indicating a "fair" agreement exists between the midwife's and the surgeon's physical examination. Sensitivity, specificity, positive and negative predictive values of "midwife's physical examination" to detect abnormal breast masses in comparison to "surgeon's physical examination" as the gold standard were 75, 67, 48, and 87 percent respectively. The results of this study do not justify the replacement of general surgeons by midwives in the health care system as the first examiner for clinical breast screening. Decisions about this issue need more comprehensive studies considering cost-effectiveness and training procedures.


Kinetic characterization and immunodetection of ecto-ATP diphosphohydrolase (EC 18.104.22.168) in cultured hippocampal neurons.
Extracellular ATP and adenosine modulate synaptic transmission in hippocampal neurons. ATP released from neural cells is hydrolyzed to adenosine by a chain of ecto-nucleotidases. ATP diphosphohydrolase hydrolyses ATP and ADP nucleotides to AMP and 5'-nucleotidase hydrolyses AMP to adenosine. In this work, we investigated the ATPase and ADPase activities of ATP diphosphohydrolase in cultured hippocampal neurons. The apparent Michaelis-Menten constant (K(m)) was 233.9 +/- 14.6 and 221.8 +/- 63.6 microM, with a calculated maximal velocity (V(max), approximately) of 49.2 +/- 10.7 and 10.9 +/- 5.2 nmol Pi/mg protein/min for ATP and ADP, respectively. The horizontal straight line obtained in the competition plot indicated that only one active site is able to hydrolyze both substrates. Furthermore, we detected the presence of this enzyme using anti-CD39 antibody, which strongly stained the soma of pyramidal and bipolar neurons, but the neurites connecting the cell clusters were also immunopositive. This antibody recognized three bands with a molecular mass close to 95, 80 and 60kDa in immunoblotting analysis. The present results show, for the first time, the kinetic and immunocytochemical characterization of an ATP diphosphohydrolase in cultured hippocampal neurons. Probably, the widespread distribution of this enzyme on the surface of neurons in culture could reflect its functional importance in studies of synaptic plasticity hippocampal.


Assessing the accuracy of conflicting autobiographical memories.
We examined criteria that rememberers and neutral audiences use to validate conflicting memories. In Experiment 1, rememberers described an incident that they recalled differently from someone else from their own and the other person's perspective. Rememberers and audiences then evaluated the accuracy of statements in both accounts, explained their accuracy ratings, and appraised the qualities of the memories. In Experiment 2, dyads who possessed conflicting memories of a shared past episode evaluated each other's recollections. In both studies, rememberers rated their own recollections as more accurate than other people's and based their judgments primarily on the internal coherence of the memories. Rememberers used consensus more frequently and sensory detail less often to evaluate other people's memories than their own. Audiences (Experiment 1) reported examining the consistency between the accounts and their own experiences; they also used consensus and normative behavior to a greater extent than rememberers. In both studies, rememberers judged their own accounts to be more detailed and coherent. Their ratings of detail and coherence predicted their ratings of accuracy.