Establishment of the block against sperm penetration in parthenogenetically activated bovine oocytes matured in vitro.
The ability of a single electric pulse to mimic a block against sperm penetration in bovine oocytes matured in vitro was investigated. Confocal laser scanning microscopy detected a global loss of spots, presumed to be cortical granules, stained with Lens culinaris agglutinin, in pulsed oocytes. Transmission electron microscopy revealed that cortical granule exocytosis occurred within 1 min of stimulation and the number of remaining cortical granules was significantly reduced in all pulsed oocytes. The ability of pulsed oocytes to undergo fertilization in vitro was also affected, as only 31% of the pulsed oocytes were penetrated compared with 87% in the control group. Since incidences of penetration in pulsed oocytes (31%), and of polyspermy in control oocytes (18%) did not differ and were highly correlated (P = 0.009) among trials (n = 15), the induced block is considered to be comparable with the natural block triggered by a spermatozoon. The increased resistance of the zona pellucida to pronase E observed in pulsed oocytes suggests that the induced block depends, at least partly, on modifications of zona pellucida glycoproteins. Finally, the majority (66%) of pulsed, penetrated oocytes did not form male pronuclei, probably as a consequence of asynchrony between the formation of female pronucleus and sperm penetration. The reduced ability of the cytoplasm to induce the formation of a male pronucleus was accompanied by a fall in histone H1 kinase activity to basal values by 3 h after stimulation. These results demonstrate that a single electric pulse can induce a block against sperm penetration similar to that of the spermatozoon itself.


Electric field driven jetting: an emerging approach for processing living cells.
This paper reports for the first time the ability to process living cellular materials by means of electrified jets at electric field strengths of up to 2 kV/mm. Bio-suspensions containing living human Jurkat cells at different concentrations were processed via this jetting approach. The jetting process was carried out at an electric field strength between 0.67 kV/mm and 2 kV/mm, corresponding to an applied voltage of 10-30 kV between two electrodes approximately 15 mm apart. The Jurkat cells were jetted under sterile conditions, collected in petri dishes and incubated for 24 and 48 hours. During and after incubation, cells were assessed for survival and structural damage; cells were found to be unharmed and to retain their integrity under all electric field strengths examined. At all field strengths jetting took place in the unstable mode. Good correlation was observed between droplet distribution plots generated by way of laser spectroscopy and estimated values from measurements of droplet relics.


Can the emotional connotation of concepts modulate the lexico-semantic deficits in Alzheimer's disease?
Semantic memory impairments are a common symptom of Alzheimer's disease (AD) and may occur at a relatively early stage. These disturbances can be evidenced by a hyperpriming effect (greater semantic priming in AD patients than in controls). Up till now, very few studies of semantic memory have included emotionally charged concepts. Our aim was therefore to study the semantic processing of such concepts, as opposed to neutral ones, in early AD. Given that emotional processes are relatively preserved at the beginning of the disease compared with other cognitive functions, we expected that an emotional connotation would influence the spreading activation of words and affect some of the impairments in semantic processing. We administered a semantic priming task (lexical decision task) implicitly assessing semantic memory to 26 patients with AD and 26 normal controls. Primes and targets either had a semantic relationship (e.g. tiger-lion), a semantic and emotional (positive or negative) relationship (e.g. slap-smack) or no relationship at all (e.g. chair-horse), or else belonged to a word-nonword condition (e.g. window-inuly). Compared with controls, the patients showed pathological hyperpriming effects in all conditions, especially in the emotional conditions. Hyperpriming implies a deterioration in specific attributes, as it is difficult to tell two concepts apart once their distinctive attributes have been lost. These results suggest that emotional concepts, like neutral ones, lose some of their distinctive attributes in early AD, and as the emotional processes are preserved, there is greater similarity between close emotional concepts than between close neutral concepts.


Prophage Typing of Staphylococcus aureus Strains Isolated from Broiler Poultry.
Staphylococcus aureus is a well-known commensal and pathogen agent of many wild and domestic animals. A wide variety of infections can be caused by S. aureus, from suppurative skin infections to life-threatening septicemia. This study was conducted to determine the prophage typing and the pattern of antibiotic resistance of S. aureus isolated from broiler poultry before they have been slaughtered. In this study, 200 nasal and cloacal swab samples from 20 different flocks were collected for bacterial isolation. Staphylococci were identified using biochemical and molecular methods before being examined for mecA gene detections in all samples resistant to oxacillin and cefotaxime. The highest value of antibiotic resistance was observed against ciprofloxacin (94%), and the maximum value of susceptibility was to gentamicin (85%). Twenty-eight (27%) samples were resistant to oxacillin. In methicillin-resistance Staphylococcus aureus (MRSA) isolates, 5 prophage types were observed, where the SGB prophage with a frequency of 75% was identified as a dominant prophage; in isolates of S. aureus susceptible to methicillin, 8 prophage types were observed, where SGFa prophage with a frequency about 82% was the dominant prophage. The high prevalence of MRSA isolates can indicate the risk of transmission of these bacteria to the food cycle. Furthermore, existence of various prophages in these isolates can be considered a threat to public health in producing pathogenicity factors in this bacterium while also empowering other bacterial pathogenicity, even other bacterial genera.


Effects of three anthelmintic schedules on the incidence of colic in horses.
Four privately owned herds (25 to 49 animals per herd) were used in a five-year trial designed to evaluate the effect of anthelmintic schedules on the incidence of colic. These herds had been treated bi-monthly with non-ivermectin, non-benzimidazole drugs for two years before the trial. Prior parasitological studies showed that they had substantial pre-treatment faecal egg counts (900 to 2200 eggs per gramme), and that they were infected with benzimidazole-resistant cyathostomes. In Years 1 and 2 of the trial, all herds (A, B, C, D) were treated bi-monthly with non-ivermectin anthelmintics (Schedule I). In Years 3 to 5, Herd A received monthly non-ivermectin anthelmintics (Schedule II) and Herd C was treated with bi-monthly ivermectin (Schedule III). Herd B was treated with Schedule II in Years 3 and 4 and Schedule III in Year 5. Herd D was maintained on Schedule I throughout the study period. In each herd, the incidence of colic, while on Schedule I, was compared to the incidence while on Schedule II or III. The risk of colic for horses on Schedule I was 2.61 to 13.04 times that of the same horses while on Schedule II and 2.27 to 9.64 times that of the same horses while on Schedule III. In Herd D, treated according to Schedule I for five years, the incidence of colic did not vary significantly throughout the study period. More effective anthelmintic treatment schedules decreased the incidence of colic in these herds, supporting the conclusion that bimonthly treatment with non-ivermectin drugs may not maximise horses' health in all management conditions.(ABSTRACT TRUNCATED AT 250 WORDS)


Emissions implications of future natural gas production and use in the U.S. and in the Rocky Mountain region.
Enhanced prospects for natural gas production raise questions about the balance of impacts on air quality, as increased emissions from production activities are considered alongside the reductions expected when natural gas is burned in place of other fossil fuels. This study explores how trends in natural gas production over the coming decades might affect emissions of greenhouse gases (GHG), volatile organic compounds (VOCs) and nitrogen oxides (NOx) for the United States and its Rocky Mountain region. The MARKAL (MARKet ALlocation) energy system optimization model is used with the U.S. Environmental Protection Agency's nine-region database to compare scenarios for natural gas supply and demand, constraints on the electricity generation mix, and GHG emissions fees. Through 2050, total energy system GHG emissions show little response to natural gas supply assumptions, due to offsetting changes across sectors. Policy-driven constraints or emissions fees are needed to achieve net reductions. In most scenarios, wind is a less expensive source of new electricity supplies in the Rocky Mountain region than natural gas. U.S. NOx emissions decline in all the scenarios considered. Increased VOC emissions from natural gas production offset part of the anticipated reductions from the transportation sector, especially in the Rocky Mountain region.


Crystalloid body, refractile body and virus-like particles in Apicomplexa: what is in there?
The phylum of Apicomplexa comprises parasitic protozoa that share distinctive features such as the apical complex, the apicoplast, specialized cytoskeletal components and secretory organelles. Other unique cytoplasmic inclusions sharing similar features have been described in some representatives of Apicomplexa, although under different denominations. These are the crystalloid body, present for example in Cryptosporidium, Plasmodium and Cystoisospora; the refractile body in Eimeria and Lankesterella; and virus-like particles, also present in Eimeria and Cryptosporidium. Yet, the specific role of these cytoplasmic inclusions in the cell cycle of these protozoa is still unknown. Here, we discuss their morphology, possible inter-relatedness and speculate upon their function to bring these organelles back to the attention of the scientific community and promote new interest towards original research on these elusive structures.


Usability and eHealth Literacy Evaluation of a Mobile Health Application Prototype to Track Diagnostic Imaging Examinations.
In the absence of a pan-Canadian electronic health record, unnecessary duplicate diagnostic imaging (DI) examinations are sometimes ordered when a physician is not aware of prior DI or prior DI is not available. Research suggests that physicians may ask their patient to recall their DI history; however, patient recall of DI can be unreliable. As a potential solution, a patient-facing mobile health application (app) prototype was developed for users to record their DI. The app was designed to be usable and inclusive to users of all health literacy levels. The aim of this paper is to demonstrate how eHealth literacy and usability heuristics can be used during the design phases of app development.


Relationship Between Nonalcoholic Fatty Liver Disease Susceptibility Genes and Coronary Artery Disease.
Coronary artery disease (CAD) is the principal cause of death in patients with nonalcoholic fatty liver disease (NAFLD). The aim of the present study was to investigate whether NAFLD is causally involved in the pathogenesis of CAD. For this, previously reported NAFLD susceptibility genes were clustered and tested for an association with CAD in the Coronary Artery Disease Genome-Wide Replication and Meta-Analysis plus the Coronary Artery Disease Genetics (CARDIoGRAMplusC4D) Consortium data set. The role of plasma lipids as a potential mediator was explored by using data from the Global Lipids Genetics Consortium. Statistical analyses revealed that the combination of 12 NAFLD genes was not associated with CAD in 60,801 CAD cases and 123,504 controls (odds ratio [OR] per NAFLD risk allele, 1.0; 95% confidence interval [CI], 0.99-1.00). In a subsequent sensitivity analysis, a positive relationship was observed after exclusion of gene variants that are implicated in NAFLD through impaired very low-density lipoprotein secretion (i.e., microsomal triglyceride transfer protein [MTTP], patatin-like phospholipase domain containing 3 [PNPLA3], phosphatidylethanolamine N-methyltransferase [PEMT], and transmembrane 6 superfamily member 2 [TM6SF2]) (OR, 1.01; 95% CI, 1.00-1.02). Clustering of the excluded genes showed a significant negative relationship with CAD (OR, 0.97; 95% CI, 0.96-0.99). A substantial proportion of the observed heterogeneity between the individual NAFLD genes in relation to CAD could be explained by plasma lipids, as reflected by a strong relationship between plasma lipids and CAD risk conferred by the NAFLD susceptibility genes (r = 0.76; P = 0.004 for low-density lipoprotein cholesterol). Conclusion: NAFLD susceptibility genes do not cause CAD per se. The relationship between these genes and CAD appears to depend to a large extent on plasma lipids. These observations strongly suggest taking plasma lipids into account when designing a new drug to target NAFLD.


Vesicourethral dysfunction following pelvic visceral ablative surgery.
Management of vesicourethral dysfunction after a major extirpative pelvic visceral operation could be complex and difficult owing to the variety of partial and complete functional and anatomical derangements produced by the primary operation. We report our experience with 22 patients who suffered various types of vesicourethral dysfunctions after extirpative pelvic visceral surgery, 5 of whom had preoperative studies. The surgical procedures were abdominoperineal resection in 9 patients, proctocolectomy in 3, anterior resection of the rectum in 2 and radical hysterectomy in 8. All 22 patients underwent urodynamic evaluations. The abnormalities noted on the preoperative urodynamic evaluations in patients about to undergo extirpative pelvic visceral surgery suggest the need for routine preoperative assessment of the lower urinary tract for an accurate understanding of the postoperative changes.


Expression and characterization of an N-truncated form of the NifA protein of Azospirillum brasilense.
Azospirillum brasilense is a nitrogen-fixing bacterium associated with important agricultural crops such as rice, wheat and maize. The expression of genes responsible for nitrogen fixation (nif genes) in this bacterium is dependent on the transcriptional activator NifA. This protein contains three structural domains: the N-terminal domain is responsible for the negative control by fixed nitrogen; the central domain interacts with the RNA polymerase σ(54) co-factor and the C-terminal domain is involved in DNA binding. The central and C-terminal domains are linked by the interdomain linker (IDL). A conserved four-cysteine motif encompassing the end of the central domain and the IDL is probably involved in the oxygen-sensitivity of NifA. In the present study, we have expressed, purified and characterized an N-truncated form of A. brasilense NifA. The protein expression was carried out in Escherichia coli and the N-truncated NifA protein was purified by chromatography using an affinity metal-chelating resin followed by a heparin-bound resin. Protein homogeneity was determined by densitometric analysis. The N-truncated protein activated in vivo nifH::lacZ transcription regardless of fixed nitrogen concentration (absence or presence of 20 mM NH(4)Cl) but only under low oxygen levels. On the other hand, the aerobically purified N-truncated NifA protein bound to the nifB promoter, as demonstrated by an electrophoretic mobility shift assay, implying that DNA-binding activity is not strictly controlled by oxygen levels. Our data show that, while the N-truncated NifA is inactive in vivo under aerobic conditions, it still retains DNA-binding activity, suggesting that the oxidized form of NifA bound to DNA is not competent to activate transcription.


Feedback facilitates transfer of training with US Hispanic workers in a healthcare laundry linen facility.
This study aimed to increase safety knowledge and behaviour of US Hispanic custodial workers in healthcare through a culturally appropriate training and monitoring process. A single-group, repeated-measures, pre-test and post-test design was used to examine training effectiveness across four sets of behaviours with 23 Spanish-speaking workers. Small group, lecture-style training in Spanish with pictures and video resulted in significant improvements in knowledge and behaviour. However, additional analyses show that behavioural feedback was the critical component in improving safety behaviour during transfer of training. Findings from reaction, knowledge, behaviour and results measures suggest that group training and graphic feedback is culturally appropriate and effective with Hispanic workers. Further investigation is needed to understand cultural factors that facilitate effective development and delivery of safety training and feedback to US Hispanic workers.


Characteristics of the isolated apical plasmalemma and intracellular tubulovesicles of the gastric acid secreting cells: demonstration of secretagogue-induced membrane mobilization.
Separation of the gradient-purified gastric microsome into two membrane subfractions of distinct enzymatic and phospholipid composition has been achieved by mild SDS (0.033% w/v) treatment followed by sucrose gradient centrifugation of the pig and rabbit gastric microsomes. While the high-density membranes had all of the (H+,K+)-ATPase and K+-pNPPase activities and revealed a single major 100-kDa band on SDS-PAGE, the low-density membranes contained all of the 5'-nucleotidase and nearly all of the Mg2+-ATPase. In the present study, the low-density subfraction has been characterized to be derived from the apical membranes and the high-density one from the intracellular tubulovesicular membranes of the parietal cells. Such characterization was based primarily on sole dependency of the apical plasma membranes on the endogenous activator for (H+,K+)-ATPase activity, differential sensitivity of the activator (AF)-dependent and -independent (H+,K+)-ATPase on micromolar vanadate and Ca2+, specific vitamin B12 binding ability of the apical plasmalemma, phospholipid and protein profiles of the two membrane subfractions, and other parameters. The AF, mentioned previously, has recently been implicated as a cytosolic regulator of the gastric (H+,K+)-ATPase [Bandopadhyay et al. (1987) J. Biol. Chem. 262, 5664-5670]. Two different forms (i.e., AF-dependent and -independent forms) of the (H+,K+)-ATPase are suggested to be present in the tubulovesicles on the basis of differential vanadate sensitivity while the AF-dependent form alone is present in the apical membranes. The data have been discussed in terms of stimulation-induced membrane transformation characteristic of the H+-secreting epithelia including the acid-secreting cells of the stomach.


Physician reimbursement: the environment for change.
Escalation of costs for physician care services in the Medicare program has elicited measures to rein in these expenditures. These include freeze and control of physician fees, limits on reimbursement for certain procedures, and mandated assignment of clinical laboratory service charges. An important step in rationalizing physician fees lies in the resource cost-based relative value study undertaken by the American Medical Association together with Harvard University economists. The results should redress inequities inherent in the current reimbursement system and allow a more suitable distribution of funds within the profession.


Associations between body mass index trajectories in childhood and cardiovascular risk factors in adulthood.
Children with a growth trajectory of overweight have higher levels of cardiovascular disease (CVD) risk factors than children with a normal-weight trajectory. However, less is known about how trajectories of body mass index (BMI) across the rest of the BMI spectrum relate to CVD risk factors and whether adult BMI affects these associations. Our aim was to examine associations between childhood BMI trajectories and adult CVD risk factors. We included 2466 individuals with childhood weights and heights (ages 6-14) from the Copenhagen School Health Records Register and adult CVD risk factors (ages 20-81) from the Copenhagen City Heart Study. Associations between childhood BMI trajectories identified by latent class modelling and CVD risk factors were examined using generalized linear regression analyses with and without adjustment for adult BMI. Normal-weight and overweight were defined by growth references from the Centers for Disease Control and Prevention. We identified four childhood trajectories within the normal-weight spectrum and one trajectory of overweight. Compared to the trajectory with the lowest BMI level, several higher BMI trajectories were associated with worse circumference, HDL and glucose homeostasis in adulthood. The highest trajectory was additionally associated with higher total cholesterol and triglycerides. When adjusting for adult BMI, the higher BMI trajectories had lower waist circumference, blood pressure and triglycerides. Trajectories of BMI within the normal-weight range and in the overweight range are associated with a worse CVD risk profile than in the lowest BMI trajectory, and these associations are modifiable by growth after childhood.


[Depopulation: science and politics].
"This paper is structured around four propositions about population decline: (1) discussions about population decline have been plagued with confusion about the very meaning of the topic, as well as about causes, consequences and policy measures; (2) the issue is closely tied to political ideologies and cultural values; (3) demographic data and methods, and some prominent demographers, have played central roles in the debate; (4) the future of fertility is even murkier than usual, and therefore the legitimacy of concerns about population decline is not obvious." (SUMMARY IN ENG AND SPA)


Kinetics of electron recombination of dye-sensitized solar cells based on TiO2 nanorod arrays sensitized with different dyes.
The performance and electron recombination kinetics of dye-sensitized solar cells based on TiO(2) films consisting of one-dimensional nanorod arrays (NR-DSSCs) which are sensitized with dyes N719, C218 and D205, respectively, have been studied. It has been found that the best efficiency is obtained with the dye C218 based NR-DSSCs, benefiting from a 40% higher short-circuit photocurrent density. However, the open circuit photovoltage of the N719 based cell is 40 mV higher than that of the organic dye C218 and D205 based devices. Investigation of the electron recombination kinetics of the NR-DSSCs has revealed that the effective electron lifetime, τ(n), of the different dye based NR-DSSCs shows the sequence of C218 > D205 > N719. The higher V(oc) with the N719 based NR-DSSC is originated from the more negative energy level of the conduction band of the TiO(2) film. In addition, in comparison to the DSSCs with the conventional nanocrystalline particles based TiO(2) films, the NR-DSSCs have shown over two orders of magnitude higher τ(n) when employing N719 as the sensitizer. Nevertheless, the τ(n) of the DSSCs with the C218 based nanorod arrays is only ten-fold higher than that of the nanoparticles based devices. The remarkable characteristic of the dye C218 in suppressing the electron recombination of DSSCs is discussed.


Electrophoretic analysis of protoplast, vacuole, and tonoplast vesicle proteins in crassulacean Acid metabolism plants.
Protoplasts and vacuoles were isolated and purified in large numbers from the CAM plants Ananas comosus (pineapple) and Sedum telephium for protein characterization. Vacuoles were further fractionated to yield a tonoplast vesicle preparation. Polypeptides of protoplasts, vacuoles, and tonoplast vesicles were compared to whole leaf polypeptides from both plants by one-dimensional sodium dodecylsulfate-polyacrylamide gel electrophoresis. Approximately 100 vacuole polypeptides could be resolved of which 25 to 30% were enriched in the tonoplast vesicles. The proteins of protoplasts, vacuoles, and tonoplast vesicles from A. comosus were analyzed further by two-dimensional gel electrophoresis. When one-dimensional electrophoretograms of A. comosus polypeptides were stained with a glycoprotein-specific periodic acid Schiff stain, very few polypeptides appeared to be glycosylated, whereas a large number of glycosylated polypeptides were detected with a silver-based glycoprotein stain particularly in tonoplast vesicles. Analysis of the enzymic content of vacuoles from both plants indicated the presence of a variety of hydrolases, including bromelain as a major constituent of A. comosus. No substrate-specific ATPase, however, could be detected in vacuoles or tonoplast vesicles from either plant.


Circ_0039569 Competes with MARCKS for miR-133b Binding Sites to Promote the Progression of Renal Cell Carcinoma.
The dysregulation of circular RNAs (circRNAs) has been shown to be correlated with the aggressiveness of renal cell carcinoma (RCC). Hence, this study investigated the role and mechanism of circ_0039569 in RCC progression. The levels of circ_0039569, miR-133b, and MARCKS (myristoylated alanine-rich protein kinase C substrate) were detected using quantitative real-time polymerase chain reaction and Western blot. In vitro experiments were conducted by using 5-ethynyl-2'-deoxyuridine assay, colony-formation assay, Transwell assay, flow cytometry, and Western blot. The direct interactions between miR-133b and circ_0039569 or MARCKS were verified by using dual-luciferase reporter and pull-down assays. Xenograft mice models were established to conduct in vivo analysis. Circ_0039569 was highly expressed in RCC tissues and cells. Functionally, silencing of circ_0039569 suppressed cell proliferation, migration, and invasion, but induced cell apoptosis in RCC cells in vitro. Moreover, mice subcutaneous xenograft assay suggested that circ_0039569 knockdown impeded tumor growth in vivo. Mechanistically, circ_0039569 acted as a sponge for miR-133b to regulate the expression of its target MARCKS. Importantly, miR-133b inhibition or MARCKS knockdown attenuated the anticancer effects of circ_0039569 knockdown on RCC growth. Diminished circ_0039569 restrains the growth and propagation of RCC cells via miR-133b/MARCKS axis, indicating an underlying effective therapeutic target for RCC patients.


Similar endothelial glycocalyx structures in microvessels from a range of mammalian tissues: evidence for a common filtering mechanism?
The glycocalyx or endocapillary layer on the luminal surface of microvessels has a major role in the exclusion of macromolecules from the underlying endothelial cells. Current structural evidence in the capillaries of frog mesentery indicates a regularity in the structure of the glycocalyx, with a center-to-center fiber spacing of 20 nm and a fiber width of 12 nm, which might explain the observed macromolecular filtering properties. In this study, we used electron micrographs of tissues prepared using perfusion fixation and tannic acid treatment. The digitized images were analyzed using autocorrelation to find common spacings and to establish whether similar structures, hence mechanisms, are present in the microvessel glycocalyces of a variety of mammalian tissues. Continuous glycocalyx layers in mammalian microvessels of choroid, renal tubules, glomerulus, and psoas muscle all showed similar lateral spacings at ∼19.5 nm (possibly in a quasitetragonal lattice) and longer spacings above 100 nm. Individual glycocalyx tufts above fenestrations in the first three of these tissues and also in stomach fundus and jejunum showed evidence for similar short-range structural regularity, but with more disorder. The fiber diameter was estimated as 18.8 (± 0.2) nm, but we believe this is an overestimate because of the staining method used. The implications of these findings are discussed.


Cytogenetic clonality analysis in myelodysplastic syndrome: monosomy 7 can be demonstrated in the myeloid and in the lymphoid lineage.
Bone marrow and blood from three patients with myelodysplastic syndrome (MDS) and monosomy 7 were studied for cell lineage involvement of the chromosomal abnormality. Cytogenetic involvement of the myeloid and erythroid cell lineages in MDS with monosomy 7 has been shown before. Lymphoid subpopulations have also been investigated but generally with negative results. A combined technique of May-Grünwald-Giemsa (MGG) for cell cytology and interphase fluorescence in situ hybridization (FISH) using a chromosome 7 specific DNA probe was applied. Further, immunophenotype and genotype of the cells were simultaneously examined with alkaline phosphatase anti-alkaline phosphatase (APAAP) immunostaining and FISH. The monosomy 7 was found in the blasts and in all or in subpopulations of myeloid and erythroid cells. T cells (CD3+, CD5+) did not appear to be involved. B cells (CD19+, CD22+) showed a normal distribution of FISH spots in two patients. In one patient however the loss of a chromosome 7 was found in approximately 70% of the cells positive for B cell markers including CD79a. The results of this study show that in some cases MDS is a disease arising in a progenitor cell with repopulative abilities restricted to myelopoiesis and erythropoiesis. In other cases, the pluripotent progenitor cells in MDS may show the capacities to differentiate into B lineage lymphoid cells, as well as suggesting that in those instances MDS represents a condition of more primitive transformed hematopoietic ancestor cells.


Measurement of Particle Concentration by Multifrequency Ultrasound Attenuation in Liquid-Solid Dispersion.
In mineral transportations, it is essential to measure the gas hydrate particle concentration to manage the risk of flowline blockage. Traditional single-frequency ultrasonic methods measure the particle concentration by treating the mixtures with an average particle size, which ignores the influence of the particle size distribution, and thus, measurement accuracy is limited. Therefore, this research studies the multifrequency ultrasound attenuation method to measure the particle concentration through the prior estimate of particle size distribution. First, considering the large particle size and low-density contrast characteristics of the hydrate-water dispersion, the influence of multiple scattering among particles cannot be ignored apart from the scattering attenuation caused by each particle, so the ultrasonic scattering attenuation mechanism considering multiple scattering effects is established to solve the attenuation prediction problem of the hydrate-water dispersion. Since the solution of the equation obtained by the ultrasonic attenuation model produces a Fredholm integral equation of the first kind, an inversion algorithm combining simulated annealing with genetic algorithm based on ultrasonic attenuation mechanism is proposed to solve the ill-posed problem in the inversion calculation of particle concentration. Finally, considering the characteristics of hydrate-water dispersion, the experiments were carried out with millimeter-sized acrylic spheres and saltwater as substitute materials of the hydrate-water dispersion. The results show that the method based on the multifrequency attenuation of ultrasound in the range 1-5 MHz has a good discrimination for the particle size, and the measurement error of particle concentration is less than 3% under different particle size distributions.


New therapeutic target for CNS injury? The role of retinoic acid signaling after nerve lesions.
Experiments with sciatic nerve lesions and spinal cord contusion injury demonstrate that the retinoic acid (RA) signaling cascade is activated by these traumatic events. In both cases the RA-synthesizing enzyme is RALDH-2. In the PNS, lesions cause RA-induced gene transcription, intracellular translocation of retinoid receptors, and increased transcription of CRBP-I, CRABP-II, and retinoid receptors. The activation of RARbeta appears to be responsible for neurotrophic and neuritogenic effects of RA on dorsal root ganglia and embryonic spinal cord. While the physiological role of RA in the injured nervous system is still under investigation three domains of functions are suggested: (1) neuroprotection and support of axonal growth, (2) modulation of the inflammatory reaction by microglia/macrophages, and (3) regulation of glial differentiation. Few studies have been performed to support nerve regeneration with RA signals in vivo, but a large number of experiments with neuronal and glial cell cultures and spinal cord explants point to beneficial effects of RA, so that future therapeutic approaches will likely focus on the activation of RA signaling.


Studies on rat liver nuclear DNA damaged by chemical carcinogen (3'-Me DAB) and AP DNA endonuclease. I. Purification and some properties of AP DNA endonucleases in rat liver chromatin.
Three kinds of apurinic/apyrimidinic (AP) DNA endonuclease, APcI, APcII, APcIII, were purified from rat liver chromatin through 1M KCl extraction, DEAE-trisacryl ion exchange chromatography. Sephadex G-150 gel filtration and AP DNA cellulose affinity chromatography. Activities of the purified APcI, APcII and APcIII were 62.5, 83.3 and 52.0 EU/mg of protein, respectively. Molecular weights of APcI, APcII and APcIII, each consisting of a single polypeptide, were 30,000, 42,000 and 13,000, and isoelectric points of them were 7.2, 6.3 and 6.2, respectively. Three enzymes showed different substrate specificities; APcI acted only on AP DNA, and APcII acted on both AP DNA and UV DNA, while APcIII acted on 3'-methyl-4-monomethylaminoazobenzene (3'-Me MAB) DNA adduct as well as AP DNA and UV DNA. These results indicate that three kinds of AP DNA endonuclease present in rat liver chromatin have structural and functional diversities.


National audit of patient choice in pediatric GH therapy.
To assess free patient choice for pediatric patients commencing growth hormone (GH) therapy within the UK and Republic of Ireland. A semistructured questionnaire was sent to all members of the British Society for Paediatric Endocrinology and Diabetes. Of 55 units responding, three do not commence patients on GH. The remaining 52 units included 21 from (all) historic growth centres, 13 from other tertiary centres, and 18 from district general hospitals. 46/52 units (89%) offer free patient choice, involving: demonstration of devices (n = 15), instructional DVDs (6), a combination of both (20), or other (3). Median (range) time spent choosing the GH device was 60 (25-150) min. Device demonstration involved: dialling up doses (39 units), assembling/dissembling (38), GH reconstitution (33), considering facilities provided by manufacturer (25), injecting patient (19), injecting parent (17), showing additional material (11), and cost consideration (4). Median (range) number of steps shown per unit was 5 (1-8), with correlation between numbers of manufacturers/devices and new patients commenced on GH per unit (R = 0.62 and 0.61, both p < 0.01), but not with time spent showing devices (R = 0.12 and 0.22, p > 0.05). Most units now offer some form of patient choice for new patients commencing GH therapy, although this involves several different methods, with larger units offering more manufacturers and more devices.


Exogenous lipoid pneumonia in children: A systematic review.
To describe the clinical-radiological-pathological characteristics and treatment outcomes of children with suspected exogenous lipoid pneumonia (ELP). Systematic review. We searched electronic databases and reference lists published between 1967 and 2018, restricted to non-accidental cases. Forty-four studies including 489 participants aged 1 day to 17 years from 13 countries were included. Cultural, medical, and behavioural rationale for oil-use was described. The clinical-radiological presentation varied widely. Diagnostic certainty was deemed highest if ELP was confirmed on bronchoalveolar lavage/frozen section lung biopsy with documented extracellular lipid on cytological staining and/or fat analysis. Non-tuberculous mycobacteria infection was identified in six studies: Mycobacterium fortuitum/chelonei, Mycobacterium smegmatis and Mycobacterium abscessus. Treatment comprised supportive therapy, corticosteroids, stopping oil, therapeutic lung-lavage and surgical resection. Outcomes were reported inconsistently. Paediatric ELP resulting from cultural and medical practices continues to be described globally. Preventive interventions, standardized reporting, and treatment efficacy studies for cases not averted, are lacking. Protocol registration: PROSPERO CRD42017068313.


Comparison of methods for isolating primary hepatocytes from mini pigs.
Successful porcine hepatocyte isolation is crucial for the development of bioartificial liver devices and hepatocyte transplantation. Serva collagenase NB grades are formulated collagenases that are suitable for various tissue isolation applications. N-acetylcysteine (NAC) can improve the viability of human hepatocytes. The aim of this study was to compare the effectiveness of two collagenases and effect of NAC on hepatocyte isolation from porcine liver tissue. Porcine hepatocytes were isolated using the perfusion method from Bama mini pigs assigned to the Serva NB 4 group (n=6), the Serva NB 8 group (n=6), or the NB 8+NAC group (n=6). Viability and yield were defined as fresh hepatocytes and their spheroids formation after 24-hour rocker culture in serum-free medium. Metabolic function was assessed by gene expression, albumin, and urea synthesis. All procedures resulted in successful hepatocyte isolation. Cells from the NB 8+NAC group had (97.8±1.9)% viability, which was higher than the NB 8 group with (94.4±2.4)% and the NB 4 group with (94.5±3.2)% (P<.001). The final cell yield reached (11.8±1.0)×10(9) cells in the NB 8+NAC group, compared to (9.5±2.1)×10(9) cells in the NB 8 group (P<.01) and (9.1±1.1) ×10(9) cells in the NB 4 group (P<.001). The secretion of albumin was superior in the NB 8+NAC group at a concentration of (425.8±35.3) ng/mL compared to the NB 8 group (339.1±32.6) ng/mL (P <.001) and NB 4 group (293.6±43.3) ng/mL (P <.01). The injury of hepatocytes also decreased in the NB 8+NAC group (P<.01). The data are presented as means ± SD. Formulated collagenase Serva NB 8 and NAC could improve the porcine hepatocyte isolation, resulting in higher yields of viable cells.


Inequities in the care of older adults: Identifying education gaps in geriatric medicine fellowship.
The past year amplified inequities in the care of older adults. Milestones focused on social determinants of health (SDOH) are lacking within Geriatric fellowship training. A virtual learning collaborative GERIAtrics Fellows Learning Online And Together (GERI-A-FLOAT) was developed to connect trainees nationwide. To address gaps in education around SDOH, a needs assessment was conducted to inform a curricular thread. A voluntary, anonymous survey was distributed to fellows through a broad network. We sought to understand prior curricula trainees had that were specifically focused on SDOH and older adults. Respondents prioritized topic areas for the curriculum. Seventy-five respondents completed the survey. More than 50% of participants indicated no training on homelessness, immigration, racism, or LGBTQ+ health at any level of medical training, with more than 70% having no training in sexism or care of formerly incarcerated older adults. The most commonly taught concepts were ableism, ageism, and poverty. Respondents prioritized the topic of racism, ageism, and ableism. There is a lack of consistent SDOH curricula pertaining to older adults across all levels of training. This needs assessment is guiding a curricular thread for GERI-A-FLOAT and ideally larger milestones for fellowships. The time is now to prepare future geriatricians to serve as change agents.


Efficient and accurate simulation of dynamic dielectric objects.
Electrostatic interactions between dielectric objects are complex and of a many-body nature, owing to induced surface bound charge. We present a collection of techniques to simulate dynamical dielectric objects. We calculate the surface bound charge from a matrix equation using the Generalized Minimal Residue method (GMRES). Empirically, we find that GMRES converges very quickly. Indeed, our detailed analysis suggests that the relevant matrix has a very compact spectrum for all non-degenerate dielectric geometries. Each GMRES iteration can be evaluated using a fast Ewald solver with cost that scales linearly or near-linearly in the number of surface charge elements. We analyze several previously proposed methods for calculating the bound charge, and show that our approach compares favorably.


Parasites as biological tags of fish stocks: a meta-analysis of their discriminatory power.
The use of parasites as biological tags to discriminate among marine fish stocks has become a widely accepted method in fisheries management. Here, we first link this approach to its unstated ecological foundation, the decay in the similarity of the species composition of assemblages as a function of increasing distance between them, a phenomenon almost universal in nature. We explain how distance decay of similarity can influence the use of parasites as biological tags. Then, we perform a meta-analysis of 61 uses of parasites as tags of marine fish populations in multivariate discriminant analyses, obtained from 29 articles. Our main finding is that across all studies, the observed overall probability of correct classification of fish based on parasite data was about 71%. This corresponds to a two-fold improvement over the rate of correct classification expected by chance alone, and the average effect size (Zr = 0·463) computed from the original values was also indicative of a medium-to-large effect. However, none of the moderator variables included in the meta-analysis had a significant effect on the proportion of correct classification; these moderators included the total number of fish sampled, the number of parasite species used in the discriminant analysis, the number of localities from which fish were sampled, the minimum and maximum distance between any pair of sampling localities, etc. Therefore, there are no clear-cut situations in which the use of parasites as tags is more useful than others. Finally, we provide recommendations for the future usage of parasites as tags for stock discrimination, to ensure that future applications of the method achieve statistical rigour and a high discriminatory power.


A global study of the unmet need for glycemic control and predictor factors among patients with type 2 diabetes mellitus who have achieved optimal fasting plasma glucose control on basal insulin.
This study used data from different sources to identify the extent of the unmet need for postprandial glycemic control in patients with type 2 diabetes mellitus (T2DM) after the initiation of basal insulin therapy in Europe, Asia Pacific, the United States, and Latin America. Different levels of evidence were used as available for each country/region, with data extracted from seven randomized controlled trials (RCTs), three clinical trial registries (CTRs), and three electronic medical record (EMR) databases. Glycemic status was categorized as "well controlled" (glycated hemoglobin [HbA1c ] at target [<7%]), "residual hyperglycemia" (fasting plasma glucose [FPG] but not HbA1c at target [FPG <7.2/7.8 mmol/L, <130/140 mg/dL, depending on country-specific recommendations]), or "uncontrolled" (both FPG and HbA1c above target). Predictor factors were identified from the RCT data set using logistic regression analysis. RCT data showed that 16.9% to 28.0%, 42.7% to 54.4%, and 16.9% to 38.1% of patients with T2DM had well-controlled glycemia, residual hyperglycemia, and uncontrolled hyperglycemia, respectively. In CTRs, respective ranges were 21.8% to 33.6%, 31.5% to 35.6%, and 30.7% to 46.8%, and in EMR databases were 4.4% to 21.0%, 23.9% to 31.8%, and 53.6% to 63.8%. Significant predictor factors of residual hyperglycemia identified from RCT data included high baseline HbA1c (all countries/regions except Brazil), high baseline FPG (United Kingdom/Japan), longer duration of diabetes (Brazil), and female sex (Europe/Latin America). Irrespective of intrinsic differences between data sources, 24% to 54% of patients with T2DM globally had residual hyperglycemia with HbA1c not at target, despite achieving FPG control, indicating a significant unmet need for postprandial glycemic control.


Tracking a hospital outbreak of KPC-producing ST11 Klebsiella pneumoniae with whole genome sequencing.
An outbreak of carbapenem-resistant Klebsiella pneumoniae strains emerged at a hospital, and was tracked in order to understand the spread of these infectious pathogens. A total of 66 K. pneumoniae strains were collected from sterile samples in 2012. The MICs of 20 antimicrobial agents were determined for all strains. Molecular typing was performed with pulsed-field gel electrophoresis (PFGE). Twelve blaKPC-producing K. pneumoniae strains isolated from ten patients were selected for whole genome sequencing. Phylogenetic reconstruction of these 12 strains was performed by the use of single-nucleotide polymorphism (SNP) row sequences of each draft genome sequence. Plasmids from the 12 strains were separated by S1 digestion and PFGE. The 12 K. pneumoniae strains isolated from the ten patients were deemed to be representative of the hospital outbreak, owing to their similar PFGE patterns. These 12 blaKPC-producing strains conferred multidrug resistance, which contrasted with the remaining 54, more susceptible, strains in the hospital. Differences in SNPs between each draft genome of the blaKPC-producing strains partitioned the 12 outbreak strains into three separate clades. The patients with each clade shared close hospital units. All 12 strains harboured at least one multidrug resistance plasmid. Strains showing high-level resistance may facilitate nosocomial dissemination and result in an infectious pathogen outbreak. Although the 12 blaKPC-producing K. pneumoniae strains possessed similar PFGE patterns, SNP variations throughout the genome allowed the strains to be divided into three clades. These results suggest that three independent transmission events led to hospital-wide dissemination of the outbreak strains.


Characterization and Function of a Novel Welan Gum Lyase From Marine Sphingomonas sp. WG.
Welan gum, a kind of microbial exopolysaccharides, produced by the genus Sphingomonas, have great potential for application in many fields, such as the food industry, cement production, and enhanced oil recovery. But there are still challenges to reduce the cost, enhance the production and the quality. Herein, the bioinformatics analysis of WelR gene was preformed, and the characterization and function of WelR, welan gum lyase, from Sphingomonas sp. WG were investigated for the first time. The results indicated that 382nd (Asn), 383rd (Met), 494th (Asn), and 568th (Glu) were the key amino acid residues, and C-terminal amino acids were essential to keeping the stability of WelR. The optimal temperature and pH of the enzymatic activity were found to be 25°C and 7.4, respectively. And WelR was good low temperature resistance and alkali resistant. K+, Mg2+, Ca2+, Mn2+, and EDTA increased WelR activities, in contrast to Zn2+. Coupled with the change in glucose concentration and growth profile, the qRT-PCR results indicated that WelR may degrade welan gum existing in the culture to maintain bacterial metabolism when glucose was depleted. This work will lay a theoretical foundation to establish new strategies for the regulation of welan gum biosynthesis.


Acute phosphate depletion dissociates hormonal stimulated second messengers in osteoblast-like cells.
The acute effect (24 h) of either phosphate depletion or phosphate surfeit on hormonal stimulated signal transduction systems was studied in the osteoblastic cell line UMR-106. Elevation of intracellular Ca2+ ([Ca2+]in), induced by different calciotropic hormones (PTH, prostaglandin E2, endothelin) was blunted by acute phosphate depletion, whereas at high inorganic phosphate (Pi) concentrations the rise in [Ca2+]in was augmented. Basal [Ca2+]in was not altered by either Pi depletion or Pi excess. The effect of acute phosphate depletion on hormonal mediated [Ca2+]in rise was not observed in the absence of extracellular Ca2+ suggesting that under these conditions, the release of Ca2+ from intracellular stores, is not affected. Also, nonhormonal calcium entry pathways such as depolarization-activated calcium channels or protein kinase C-activated Ca2+ channels were not affected by acute phosphate depletion. cAMP accumulation in the cells, either through receptor or nonreceptor-mediated mechanisms, increased under low Pi conditions and decreased as Pi concentration in the culture media was progressively increased from 0 to 2 mM during 24 h of incubation. Changes in Pi concentration had no effect on basal cAMP generation by the cells. The facilitative effect of acute Pi depletion on agonist-induced cAMP accumulation could be demonstrated in both the presence and absence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (0.2 mM). PTH receptor binding assessed with [Nle8 Nle18 Tyr34] bovine PTH (1-34) NH2 was not altered by phosphate depletion. We conclude that exposure of osteoblasts to different Pi environments modulates the second messenger responses to hormones in a reciprocal fashion so that acute phosphate depletion down-regulates [Ca2+]in signals while augmenting cAMP generation and vice versa. Inasmuch as bone resorption processes can be modulated by Ca2+ and cAMP the data presented herein suggest that the altered bone resorptive response to calciotropic hormones (e.g. PTH), under surfeit or deficit of phosphate, is mediated by changes in [Ca2+]in and cAMP.


White Matter Microstructure Changes and Cognitive Impairment in the Progression of Chronic Kidney Disease.
Cognitive impairment is a well-defined complication of chronic kidney disease (CKD), but the neural mechanisms are largely unknown. The study aimed to assess white matter (WM) microstructure changes and their relationship with cognitive impairment development during CKD progression. Diffusion tensor imaging (DTI) datasets were acquired from 38 patients with CKD (19 patients were at stage 3; 19 patients were at stage 4) and 22 healthy controls (HCs). Tract-based spatial statistics (TBSS) was implemented to assess the differences in WM integrity among the three groups. The associations between abnormal WM integrity and clinical indicators (digit symbol test scores, the type A number connection test scores, hemoglobin, serum urea, serum creatinine, serum calcium, and serum potassium levels) were also computed. Compared with patients with CKD at stage 3 and HCs, patients with CKD at stage 4 showed significantly lower fractional anisotropy (FA) and higher mean diffusivity (MD) in the corpus callosum (CC), anterior thalamic radiation, inferior fronto-occipital fasciculus, and inferior longitudinal fasciculus. Correlation analysis showed that the MD in the genu of CC was negatively associated with the digit symbol test scores (r = -0.61, p = 0.01), and the FA in the left anterior thalamic radiation was positively associated with the level of serum calcium (r = 0.58, p = 0.01). Patients with non-end-stage CKD have multiple abnormalities in WM regions. DTI metrics change with the progression of CKD and are primarily associated with cognitive impairment. The reduced integrity of WM tracts may be related to a low level of blood calcium.


Pathogenesis of Campylobacter fetus infections: critical role of high-molecular-weight S-layer proteins in virulence.
Wild-type Campylobacter fetus strains possess high-molecular-weight S-layer proteins (S+) and are highly resistant to serum-mediated killing and phagocytosis. Spontaneous mutant strains lacking these proteins (S-) are serum and phagocytosis sensitive and have reduced virulence in a mouse model. Intact S+ cells were treated with pronase, which made them S- although genotypically S+ and had essentially no effect on other cellular proteins or on viability. Treatment with pronase, but not buffer alone, rendered these cells serum and phagocytosis sensitive and reduced mouse virulence to the level observed for the S- mutant cells. In related studies, purified S-layer proteins diminished neutrophil chemoluminescent responses to a heterologous particulate antigen. Finally, passive administration of antiserum to the 97-kDa S-layer protein partially protected mice against lethal challenge with the S+ strain. These studies define the contribution of the S-layer proteins to C. fetus virulence.


Preparation of Ni-based metal monolithic catalysts and a study of their performance in methane reforming with CO2.
A series of Ni/SBA-15/Al(2)O(3)/FeCrAl metal monolithic catalysts with Ni loadings varying between 3 % and 16 % were prepared, and their structure was characterized by various techniques. The catalytic activity of the catalyst for methane reforming with CO(2) leading to synthesis gas was evaluated using a fixed-bed reactor. The results indicate good catalytic activity of the Ni/SBA-15/Al(2)O(3)/FeCrAl samples under the reaction conditions. The catalyst with a Ni loading of 8.0 % displays excellent activity and stability at 800 degrees C over 1400 h time on stream. After reaction, the hexagonal mesoporous structure of SBA-15 is still present and the pore walls of SBA-15 prevent the aggregation of nickel. Interactions between NiO, SBA-15, and the Al(2)O(3)/FeCrAl support modify the redox properties of the Ni/SBA-15/Al(2)O(3)/FeCrAl catalysts.


Related factor analysis for predicting large-volume central cervical lymph node metastasis in papillary thyroid carcinoma.
The aim of this study was to investigate the factors related to large-volume central cervical lymph node metastasis (LNM) in papillary thyroid carcinoma. A retrospective study of 340 patients with 642 papillary thyroid carcinoma nodules who underwent thyroidectomy in Peking Union Medical College Hospital between 2011 and 2015 was conducted. These nodules were divided into two groups by the number of central cervical lymph node metastases: large-volume central cervical LNM (>5 metastatic lymph nodes, n = 129) and no central cervical LNM (n = 211). We evaluated the correlations between gender, age, chronic lymphocytic thyroiditis, thyroid ultrasonographic features, and large-volume central cervical LNM. We found that younger age (≤40 years) (OR = 3.796, 95% CI = 2.842, 5.070), male gender (OR = 4.005, 95% CI = 2.858, 5.61), and ultrasonographic features such as tumor macroaxis size (OR = 2.985, 95% CI = 1.581, 5.633), tumor located in the isthmus (OR = 7.578, 95% CI = 4.863, 11.810), ill-defined margin (OR = 3.008, 95% CI = 1.986, 4.556), microcalcification (OR = 2.155, 95% CI = 1.585, 2.929), and abnormal cervical lymph nodes (OR = 13.753, 95% CI = 9.278, 20.385) were independent risk factors for large-volume central cervical LNM in papillary thyroid carcinoma, while chronic lymphocytic thyroiditis (OR = 0.248, 95% CI = 0.172, 0.358) was a protective factor. Younger age (≤40 years), male sex, and ultrasonographic features such as tumor macroaxis size, tumor located in the isthmus, ill-defined margin, microcalcification, and abnormal cervical lymph nodes were independent risk factors for large-volume central cervical LNM in papillary thyroid carcinoma, while chronic lymphocytic thyroiditis can be considered a protective factor. Our results provide a reference for adjusting clinical treatment approaches.


A Simplified Calculation Method of Heat Source Model for Induction Heating.
Line heating is used in forming the complex curve plates of ships, and this process is becoming integrated into automated tools. Induction heating equipment has become commonly used in automatic line heating. When applying automated equipment, it is necessary to calculate the relationship between the heating parameters and the temperature field. Numerical methods are primarily used to accomplish the calculations for induction heating. This computation process requires repeated iterations to obtain a stable heat generation rate. Once the heat generation rate changes significantly, a recalculation takes place. Due to the relative position of the coil and plate changes during heating, the grid needs to be frequently re-divided during computation, which dramatically increases the total computation time. In this paper, through an analysis of the computation process for induction heating, the root node that restricts the computation efficiency in the conventional electromagnetic-thermal computation process was found. A method that uses a Gaussian function to represent the heat flux was proposed to replace the electromagnetic computation. The heat flux is the input for calculating the temperature field, thus avoiding the calculation of the electromagnetic analysis during induction heating. Besides, an equivalence relationship for multi-coil was proposed in this paper. By comparing the results of the experiment and the numerical method, the proposed heat source model's effectiveness was verified.


Role of cyclic AMP response element binding protein (CREB) in angiotensin II-induced responses in vascular smooth muscle cells.
Cyclic AMP response element (CRE) binding protein (CREB) is a nuclear transcription factor that regulates the transcription of several genes containing the CRE sites on their promoters. CREB is activated by phosphorylation on a key serine residue, Ser311, in response to a wide variety of extracellular stimuli including angiotensin II (Ang II). Ang II is an important vasoactive peptide and mitogen for vascular smooth muscle cells (VSMC) that in addition to regulating the contractile response in VSMC also plays an important role in phenotypic switch of VSMC from contractile to a synthetic state. The synthetic VSMC are known to exhibit proliferative and migratory properties due to hyperactivation of Ang II-induced signaling events. Ang II has been shown to induce CREB phosphorylation/activation and transcription of genes implicated in proliferation, growth, and migration. Here, we have highlighted some key studies that have demonstrated an important role of CREB in Ang II-mediated gene transcription, proliferation, hypertrophy, and migration of VSMC.


Amino Acid Profile in Malnourished Patients with Liver Cirrhosis and Its Modification with Oral Nutritional Supplements: Implications on Minimal Hepatic Encephalopathy.
Low plasma levels of branched chain amino acids (BCAA) in liver cirrhosis are associated with hepatic encephalopathy (HE). We aimed to identify a metabolic signature of minimal hepatic encephalopathy (MHE) in malnourished cirrhotic patients and evaluate its modification with oral nutritional supplements (ONS) enriched with ß-Hydroxy-ß-methylbutyrate (HMB), a derivative of the BCAA leucine. Post hoc analysis was conducted on a double-blind placebo-controlled trial of 43 individuals with cirrhosis and malnutrition, who were randomized to receive, for 12 weeks, oral supplementation twice a day with either 220 mL of Ensure® Plus Advance (HMB group, n = 22) or with 220 mL of Ensure® Plus High Protein (HP group, n = 21). MHE evaluation was by psychometric hepatic encephalopathy score (PHES). Compared to the HP group, an HMB-specific treatment effect led to a larger increase in Val, Leu, Phe, Trp and BCAA fasting plasma levels. Both treatments increased Fischer's ratio and urea without an increase in Gln or ammonia fasting plasma levels. MHE was associated with a reduced total plasma amino acid concentration, a reduced BCAA and Fischer´s ratio, and an increased Gln/Glu ratio. HMB-enriched ONS increased Fischer´s ratio without varying Gln or ammonia plasma levels in liver cirrhosis and malnutrition, a protective amino acid profile that can help prevent MHE.


Working Memory Load and Reminder Effect on Event-Based Prospective Memory of High- and Low-Achieving Students in Math.
The effects of working memory (WM) demand and reminders on an event-based prospective memory (PM) task were compared between students with low and high achievement in math. WM load (1- and 2-back tasks) was manipulated as a within-subject factor and reminder (with or without reminder) as a between-subject factor. Results showed that high-achieving students outperformed low-achieving students on all PM and n-back tasks. Use of a reminder improved PM performance and thus reduced prospective interference; the performance of ongoing tasks also improved for all students. Both PM and n-back performances in low WM load were better than in high WM load. High WM load had more influence on low-achieving students than on high-achieving students. Results suggest that low-achieving students in math were weak at PM and influenced more by high WM load. Thus, it is important to train these students to set up an obvious reminder for their PM and improve their WM.


[The angiography of kidney transplant arteries using carbon dioxide].
After renal transplantation hypertension occurs in 60-80% of patients, in 3-23% of them due to renal artery stenosis in the transplanted kidney. Angiographic assessment of the severity of the stenosis would best be done with a non-nephrotoxic contrast medium. We investigated the use of carbon dioxide, a bubble-forming contrast medium excreted via the lung, for its suitability in the angiography of the arterial system in the renal transplant. In an uncontrolled series of observations on six patients with renal transplants and hypertension who were being treated with three or four antihypertensive drugs, the transplant's arteries were demonstrated with carbon dioxide, injected by Seldinger technique into the ipsilateral femoral artery, and the results recorded by subtraction angiography. The angiograms demonstrated postoperative kinked renal artery, a plaque in the external iliac artery, and a sharply angled origin of the external iliac artery in one case each, and three normal cases. Carbon dioxide is a suitable medium for demonstrating renal arteries in a transplanted kidney and is preferable to conventional contrast media for the angiographic diagnosis of possible renal artery stenosis.


[What is the onset of primary biliary liver cirrhosis?].
The medical records of 49 consecutive patients with primary biliary cirrhosis of the liver were screened for informations about medical examinations during the years before the diagnosis was established. In 15 cases previous medical reports could be found. Evidence of liver disease (slight elevation of transaminases and gamma-GT) was documented up to 18 years before the diagnosis was proven. In 6 patients liver biopsies had been performed: normal 1 x, fatty liver 1 x, fibrosis 1 x, non-specific hepatitis 1 x, chron. pers. Hep. 2 x. The characteristic increase of alkaline phosphatase often occurred within a few months. Antimitochondrial antibodies became positive independent of the beginning of cholestasis. It can be concluded that early stage primary biliary cirrhosis must be considered in patients with long standing slight elevation of liver enzymes even without cholestasis when other causes can be excluded.


TP53 mutations detected in circulating tumor cells present in the blood of metastatic triple negative breast cancer patients.
Circulating tumor cells (CTCs) are tumor cells shed from either primary tumors or its metastases that circulate in the peripheral blood of patients with metastatic cancers. The molecular characterization of the CTCs is critical to identifying the key drivers of cancer metastasis and devising therapeutic approaches. However, the molecular characterization of CTCs is difficult to achieve because their isolation is a major technological challenge. CTCs from two triple negative breast cancer patients were enriched using CellSearch and single cells selected by DEPArray™. A TP53 R110 fs*13 mutation identified by next generation sequencing in the breast and chest skin biopsies of both patients was studied in single CTCs. From 6 single CTC isolated from one patient, 1 CTC had TP53 R110 delC, 1 CTC showed the TP53 R110 delG mutation, and the remaining 4 single CTCs showed the wild type p53 sequence; a pool of 14 CTCs isolated from the same patient also showed TP53 R110 delC mutation. In the tumor breast tissue of this patient, only the TP53 R110 delG mutation was detected. In the second patient a TP53 R110 delC mutation was detected in the chest wall skin biopsy; from the peripheral blood of this patient, 5 single CTC and 6 clusters of 2 to 6 CTCs were isolated; 3 of the 5 single CTCs showed the TP53 R110 delC mutation and 2 CTCs showed the wild type TP53 allele; from the clusters, 5 showed the TP53 R110 delC mutation, and 1 cluster the wild type TP53 allele. Single white blood cells isolated as controls from both patients only showed the wild type TP53 allele. We are able to isolate uncontaminated CTCs and achieve single cell molecular analysis. Our studies showed the presence of different CTC sub-clones in patients with metastatic breast cancer. Some CTCs had the same TP53 mutation as their matching tumor samples although others showed either a different TP53 mutation or the wild type allele. Our results indicate that CTCs could represent a non-invasive source of cancer cells from which to determine genetic markers of the disease progression and potential therapeutic targets.


Chronic follicular bronchiolitis requires antigen-specific regulatory T cell control to prevent fatal disease progression.
To study regulatory T (Treg) cell control of chronic autoimmunity in a lymphoreplete host, we created and characterized a new model of autoimmune lung inflammation that targets the medium and small airways. We generated transgenic mice that express a chimeric membrane protein consisting of hen egg lysozyme and a hemoglobin epitope tag under the control of the Clara cell secretory protein promoter, which largely limited transgene expression to the respiratory bronchioles. When Clara cell secretory protein-membrane hen egg lysozyme/hemoglobin transgenic mice were crossed to N3.L2 TCR transgenic mice that recognize the hemoglobin epitope, the bigenic progeny developed dense, pseudo-follicular lymphocytic peribronchiolar infiltrates that resembled the histological pattern of follicular bronchiolitis. Aggregates of activated IFN-γ- and IL-17A-secreting CD4(+) T cells as well as B cells surrounded the airways. Lung pathology was similar in Ifng(-/-) and Il17a(-/-) mice, indicating that either cytokine is sufficient to establish chronic disease. A large number of Ag-specific Treg cells accumulated in the lesions, and Treg cell depletion in the affected mice led to an interstitial spread of the disease that ultimately proved fatal. Thus, Treg cells act to restrain autoimmune responses, resulting in an organized and controlled chronic pathological process rather than a progressive disease.


Effect of dietary bulk on organ mass, fasting heat production and metabolism of the small and large intestines in sheep.
Groups of 10 lambs were fed either a control diet or a diet consisting of 10% polyethylene at isoenergetic levels for 30 d to study the effect of dietary bulk on visceral organ mass and metabolism of mucosa from the jejunum and proximal colon. Fasting heat production was measured using an additional six lambs. Weights of the large intestine, stomach complex and lungs were greater in lambs fed the polyethylene diet than control lambs. Liver and small intestine weights were not affected by diet. Oxygen consumption rates of jejunal mucosa were not affected by diet while a trend existed for oxygen consumption rates of mucosa from the proximal colon to be increased due to feeding polyethylene. Na+,K+ ATPase activity of mucosa from both jejunum and colon was increased due to feeding polyethylene. Fasting heat production, oxygen consumption or carbon dioxide production did not differ with diet. Dietary bulk affects growth and metabolism of visceral organs but metabolic modifications appear different among tissues.


The origin of a Robertsonian chromosomal translocation in house mice inferred from linked microsatellite markers.
The western European house mouse, Mus domesticus, includes many distinct Robertsonian (Rb) chromosomal races. Two competing hypotheses may explain the distribution of Rb translocations found in different populations: they may have arisen independently multiple times, or they may have arisen once and been spread through long-distance dispersal. We investigated the origin of the Rb 5.15 translocation using six microsatellite loci linked to the centromeres of chromosomes 5 and 15 in 84 individuals from three Rb populations and four neighboring standard-karyotype populations. Microsatellite variation on the 5.15 metacentric chromosomes was significantly reduced relative to the amount of variation found on acrocentric chromosomes 5 and 15, suggesting that linked microsatellite loci can track specific mutational events. Phylogenetic analyses resulted in trees which are consistent with multiple origins of the 5.15 metacentric chromosomes found in the three Rb populations. These results suggest that cytologically indistinguishable mutations have arisen independently in natural populations of house mice.


Wild ungulates as sentinel of BTV-8 infection in piedmont areas.
Bluetongue caused by the genotype 8 virus (BTV-8) appeared for the first time in BTV free areas in northern Italy in 2008. The presence of domestic animals outbreaks, abundant wild ungulates populations, and ongoing regional BTV control plans, made this area interesting to evaluate the role of wild ruminants in BTV-8 epidemiology. We analyzed spleen samples from hunted red deer (Cervus elaphus), roe deer (Capreolus capreolus) and Alpine chamois (Rupicapra rupicapra) by quantitative RT-PCR. Samples were collected from 2008 to 2011 in two provinces of Piedmont region. BTV-8 was detected in all ungulate species, confirming their receptivity to the infection. However, the viral load in the positive specimens was low, and decreased from 2008 to 2011. These results, together with the extinction of the epidemic following a regional livestock vaccination campaign, lead to hypothesize that wild ungulates were an epiphenomenon and they had not an important role in the domestic transmission cycle of BTV-8 in this area. In spite of this, wild ruminants appear to be good sentinels of BTV circulation and their monitoring could be useful for surveillance in piedmont areas.


[MR imaging of the shoulder. Impingement and instability-related abnormalities].
In addition to the case history and the clinical examination, MR imaging has an important role in the diagnosis and differential diagnosis of numerous shoulder abnormalities and in the investigation of chronic shoulder pain. Important indications for MR imaging are any conditions or symptoms making assessment of the rotator cuff and the labrocapsular complex necessary. Assessment of the rotator cuff muscles, in particular, is crucial. The value of MR arthrography, which is still controversial, is discussed. The greatest potential benefit of MR arthrography is the accurate evaluation of subtle rotator cuff abnormalities and shoulder instability-related lesions, and the assessment of pathologic conditions of the long biceps tendon. This paper describes the most common pathologic findings of the shoulder joint and describes how the relevant findings are reported and quantified for the orthopaedic shoulder surgeon.


Morphological and Developmental Features of Stone Cells in Eriobotrya Fruits.
Some members of the Rosaceae family, particularly pear, contain stone cells in their fruits. Although stone cells in pear fruits are well studied, relatively little attention has been given to loquat stone cells. Only a few reports have suggested a relationship between stone cell traits and storage and transport tolerance of loquat fruits. Previously, we generated the variety JT8 from the interspecific hybrid of the loquat cultivar Jiefangzhong (JFZ; Eriobotrya japonica Lindl. cv. Jiefangzhong, female parent) and wild Taiwanese loquat (TL; E. deflexa Nakai, male parent). The JT8 fruits had a granular feel, similar to that of pear fruits, due to the presence of stone cells. In this study, the shape, size, development, and distribution dynamics of stone cells of Eriobotrya plants were thoroughly investigated. The results showed that loquat stone cells are brachysclereids and often contain typical branching pits. Loquat stone cells were distributed as both single stone cells and in stone cell clusters (SCCs), and the density of the stone cells near the core was higher than that near the peel. Stone cell density first increased and then decreased during fruit development. These traits noted in Eriobotrya were very similar to those observed in pear, indicating a close relationship between loquat and pear. Moreover, the contents, density dynamics, and aggregation traits of stone cells of the interspecific hybrid JT8 were derived from the male parent (TL). Transgressive segregation was likely exhibited in the content of stone cells and the size of the SCCs. More specifically, the content of stone cells reached 1.61% (w/w). In extreme cases, SCCs of JT8 exceeded 1,000 μm in length and 500 μm in width. This demonstrated that stone cell traits could be transmitted from parent to progeny through interspecific hybridization. The density dynamics of stone cells in two loquat cultivars with different storage and transport tolerances were also investigated, which indicated that the cultivar with more stone cells was more tolerant to storage and transport. We suggest that wild loquat genetic resources containing stone cells in Eriobotrya plants can be used to gradually improve the storage and transport tolerance of loquat fruits.


AGA Clinical Practice Update on Approach to the Use of Noninvasive Colorectal Cancer Screening Options: Commentary.
The purpose of this American Gastroenterological Association (AGA) Institute Clinical Practice Update Commentary is to review the available evidence and provide expert advice regarding the approach to using noninvasive colorectal cancer (CRC) screening options, including evidence for their effectiveness, selection of individuals for whom these tests are appropriate, implications of a positive non-colonoscopy screening test, and opportunities to enhance the quality of noninvasive CRC screening programs. This Clinical Practice Update was commissioned and approved by the AGA Institute Clinical Practice Updates Committee and the AGA Governing Board to provide timely guidance on a topic of high clinical importance to the AGA membership, and underwent internal peer review by the Clinical Practice Updates Committee and external peer review through standard procedures of Gastroenterology. This expert commentary reflects recently published studies in this field, as well as the experiences of the authors who are gastroenterologists with high-level expertise in CRC screening and prevention.


The role of psychosocial interventions in bipolar II disorder: what do we know?
The tendency for patients with bipolar II disorder to present with depressive symptoms rather than hypomanic symptoms can lead to a misdiagnosis of unipolar depression. These patients are often treated with antidepressants, which may be inappropriate for patients with bipolar II disorder due to the risk of inducing manic switching or rapid cycling. Misdiagnosis and mistreatment or nontreatment can lead to substantial psychosocial dysfunction, which may be best addressed in the context of psychotherapy. Psychotherapy in conjunction with pharmacotherapy has been shown to be an effective treatment for patients with bipolar II disorder, but more research is needed to better understand how treatment interventions should be used to optimize symptomatic and functional outcomes.


Tooth size discrepancies in Class II division 1 and Class III malocclusion requiring surgical-orthodontic or orthodontic treatment.
To compare mean anterior (AR) and mean overall (OR) tooth size ratios, prevalence of clinically significant tooth size discrepancies (TSDs) and correlation between AR and OR in subjects with Class II division 1 and Class III malocclusion treated by surgical-orthodontic or orthodontic means. Retrospective, cross-sectional. State-funded and private clinics. From pre-treatment cohorts of 770 surgical and 610 non-surgical subjects, Class II division 1 and Class III malocclusion groups were identified with 60 surgical and 60 non-surgical subjects, comprising 30 males and 30 females, in each. AR and OR were calculated by landmarking digital models. Differences in AR and OR and their relationship were analysed using two-way analysis of variance (ANOVA) and a correlation coefficient, respectively. The proportions of the surgical and non-surgical groups with a TSD were assessed using logistic regression. Intra-examiner reproducibility involved re-landmarking 30 randomly selected image sets and differences in ARs and ORs were compared using a paired t-test. Random error was assessed using the intraclass correlation coefficient (ICC). Analyses were performed using SAS (SAS Institute Inc., Cary, NC, USA) at the 5% level of significance. There were no statistically significant differences associated with the measurement of either the mean AR (P = 0·913) or the mean OR (P = 0·874). ICC values were very high (AR = 0·95; OR = 0·90). Differences existed between both Class II and Class III surgical (AR: P<0·001; OR: P<0·001) and non-surgical groups (AR: P = 0·012; OR: P = 0·003). The AR and OR relationship was strong (correlation coefficient = 0·72). The highest percentage of clinically significant TSDs was seen in the AR of both Class II and Class III surgical groups (23·3%). In the cohort examined: AR and OR differed significantly for malocclusion groups. The prevalence of clinically significant TSDs did not differ significantly between surgical and non-surgical groups although the highest percentage of clinically significant TSDs was recorded for AR in Class II and Class III surgical cases. AR and OR were closely related.


Cancer prevention and early detection among Egyptians.
Although cancer is frequently not perceived as a significant health problem in developing countries, cancer death in developing countries is higher than in developed countries. Therefore, cancer prevention and early detection in developing countries is becoming a universal challenge to health care providers. The purpose of this exploratory study was to identify knowledge, attitudes, and behaviors related to cancer prevention and early detection among Egyptians. Data were collected from 99 people, and the design was descriptive and correlational, using a self-administered survey. Results showed that the Egyptians surveyed had some knowledge about warning signs of cancer as well as prevention and early detection strategies. In addition, results indicated a significant difference between males and females in warning signs of cancer as well as prevention and early detection methods. Participants were less likely to believe their lifestyle behaviors affect their chances of developing cancer. Current practices undertaken included dietary behaviors, abstinence from smoking, check-up for minor suspicious symptoms, and annual complete physical examinations. Barriers to undertaking practices included factors related to lack of preventive resources, lack of information about preventive strategies, family financial concerns, and specific attitudes toward cancer. The study findings indicate the need for providing people with information to help them make decisions regarding undertaking cancer prevention and early detection practices.


Determining the burden of tuberculosis in Eritrea: a new approach.
To obtain an estimate of the prevalence of new cases of smear-positive tuberculosis in Eritrea using a new low-cost approach. The study was designed to include a sample of 35,000 people divided between 40 clusters. The clusters were selected by sampling proportional to population size. In each cluster, census data were obtained from eligible individuals. Individuals aged 15 years or more were questioned about cough and its duration and smoking and were asked to provide two samples of sputum (spot and morning) for examination by fluorescence microscopy. A total of 38,032 individuals were included in the study. Of the 19,197 individuals aged 15 years or more, 18,152 (94.6%) provided at least one sample of sputum. Fifteen individuals fulfilled the case definition, providing a prevalence of new cases of smear-positive tuberculosis of 90 per 100,000 (95% confidence interval, CI, 35-145 per 100,000) individuals aged 15 years or more and 50 per 100,000 (95% CI, 19-80 per 100,000) in the total population. The estimated prevalence of new cases of smear-positive tuberculosis obtained from our study is considerably lower than the estimate published by the World Health Organization. The new methodology for surveys to determine the prevalence of tuberculosis that we used is feasible in the field. This methodology should be validated in other countries and compared with other methods for measuring prevalence.


[Inferior mesocaval shunt].
The results of inferior mesenteric vein-inferior vena cava shunt coupled with splenectomy and gastroesophageal devascularization in 16 patients with cirrhotic portal hypertention were reviewed. The mean diameter of the shunt stoma was 6.46 +/- 0.78 mm, and the portal pressure decreased from 4.25 +/- 0.54 kPa to 3.39 +/- 0.41 kPa. Variceal bleeding stopped in all 16 patients with no severe postoperative complications. This procedure should preserve satisfactory portal perfusion to the liver, and technically is easier than other shunt procedures.


Rupture of splenic artery pseudoaneurysm.
Pseudoaneurysms of the splenic artery have been well described in association with pancreatic pseudocysts secondary to pancreatitis. We present a case of a ruptured splenic artery pseudoaneurysm 14 years after splenectomy for trauma which, at that time, was complicated by a subphrenic abscess.


Exon 3 of the growth hormone receptor (GH-R) is specific to eutherian mammals.
Growth hormone receptor (GH-R) plays a critical role in the control of growth and metabolism in all vertebrates. GH-R consists of 9 coding exons (2-10) in all eutherian mammals, while the chicken only has 8 coding exons, and does not have an orthologous region to exon 3 in eutherians. To further understand the evolutionary origins of exon 3 of the GH-R in eutherians we cloned the full-length GH-R sequence in a marsupial, the tammar wallaby to determine whether exon 3 was present or absent in marsupial liver cDNA. There was no evidence for the presence of an exon 3 containing mRNA in sequence of tammar pouch young and adult livers. We next examined the genomes of the platypus (a monotreme mammal) and the grey short-tailed opossum (another marsupial). Like the tammar, the GH-R gene of neither species contained an exon 3. GH receptor can obviously function in the absence of this exon, raising speculation about the function of this domain, if any, in eutherians. A comparison of exon 3 protein sequences within 16 species of eutherian mammals showed that there was approximately 75% homology in the domain but only 3 of the 21 amino acids were identical (Leu12, Gln13 and Pro17). Interestingly, we detected greater evolutionary divergence in exon 3 sequences from species that have variants of GH or prolactin (PRL) in their placentas. These data show that exon 3 was inserted into the GH-R after the divergence of the marsupial and eutherian lineages at least 130 million years ago.


Influence of Electromechanical Activity on Cardiac Differentiation of Mouse Embryonic Stem Cells.
During differentiation, mouse embryonic stem cell-derived cardiomyocytes (mESC-CMs) receive electromechanical cues from spontaneous beating. Therefore, promoting electromechanical activity via electrical pacing or suppressing it by drug treatment might affect the cellular functional development. Electrical pacing was applied to confluent monolayers of mESC-CMs during late-stage differentiation (days 16-18). Alternatively, spontaneous contraction was suppressed by (a) blocking ion currents with CsCl (HCN channel), trazodone (T-type Ca2+ channel), or both CsCl and trazodone on days 11-18; or (b) applying blebbistatin (excitation-contraction uncoupler) on days 11-14. Electrophysiological properties and gene expression were examined on day 19 and 18, respectively. Optical mapping revealed no significant difference in conduction velocity (CV)in paced vs. non-pacedmonolayers, nor were there significant changes in gene expression of connexin-43, Na-Ca exchanger (NCX), or myosin heavy chain (MHC). However, CV variability among differentiation batches and CV heterogeneity within individual monolayers were significantly lower in paced mESC-CMs. Alternatively, while the four drug treatments suppressed contraction with varying degrees (up to complete inhibition), there was no significant difference in CV for any of the treatments compared with controls. Trazodone treatment significantly reduced CV variability as compared to controls, whereas CsCl treatment significantly reduced CV heterogeneity. Distinct changes in gene expression of connexin-43, MHC, HCNl, Cav3.1/3.2 were not observed. Electrical pacing, but not suppression of spontaneous contraction, during late-stage differentiation reduces the intrinsic variability of CV among differentiation batches and across individual monolayers, which can be beneficial in the application of ESCs for myocardial tissue repair.


3D physiological model of the aortic valve incorporating small coronary arteries.
The diseases of the coronary arteries and the aortic root are still the leading causes of mortality and morbidity worldwide. In this study, a 3D global fluid-structure interaction of the aortic root with inclusion of anatomically inspired small coronary arteries using the finite element method is presented. This innovative model allows to study the impact and interaction of root biomechanics on coronary hemodynamics and brings a new understanding to small coronary vessels hemodynamics. For the first time, the velocity profiles and shear stresses are reported in distal coronary arteries as a result of the aortic flow conditions in a global fluid-structure interaction model.


Facial palsy due to tuberculosis: the value of CT.
This case report demonstrates the value of CT in the investigation of facial palsy due to tuberculous mastoiditis. This is the best imaging modality available for identifying features suggestive of tuberculosis, the disease extent, presence of complications and demonstration of anatomy prior to surgery.


[Nursing diagnoses identified in pregnant patients under prenatal care].
This descriptive research aimed to analyze the Nursing Diagnoses identified in low-risk pregnant patients. Eleven pregnant women received prenatal care in a safe maternity in the state of Goiás. The data were classified under the NANDA (North American Nursing Diagnosis Association) taxonomy II and analyzed according to the referrals in obstetrics. Twenty-five different Nursing Diagnoses were identified. All pregnant women presented: insufficient knowledge (different levels and subjects), disturbed sleep pattern, activity intolerance, self-care deficit (bathing and hygiene), stress urinary incontinence, and some risk diagnoses. Identifying NANDA's Nursing Diagnoses allows the identification of the risk level, early detection of complications, and individualization in nursing assistance.


Bacterial Infection and Hypersensitive Response Assays in Arabidopsis-Pseudomonas syringae Pathosystem.
Arabidopsis thaliana-Pseudomonas syringae pathosystem has been used as an important model system for studying plant-microbe interactions, leading to many milestones and breakthroughs in the understanding of plant immune system and pathogenesis mechanisms. Bacterial infection and plant disease assessment are key experiments in the studies of plant-pathogen interactions. The hypersensitive response (HR), which is characterized by rapid cell death and tissue collapse after inoculation with a high dose of bacteria, is a hallmark response of plant effector-triggered immunity (ETI), one layer of plant immunity triggered by recognition of pathogen-derived effector proteins. Here, we present a detailed protocol for bacterial disease and hypersensitive response assays applicable to studies of Pseudomonas syringae interaction with various plant species such as Arabidopsis, Nicotiana benthamiana, and tomato.


Mice with ribosomal protein S19 deficiency develop bone marrow failure and symptoms like patients with Diamond-Blackfan anemia.
Diamond-Blackfan anemia (DBA) is a congenital erythroid hypoplasia caused by a functional haploinsufficiency of genes encoding for ribosomal proteins. Among these genes, ribosomal protein S19 (RPS19) is mutated most frequently. Generation of animal models for diseases like DBA is challenging because the phenotype is highly dependent on the level of RPS19 down-regulation. We report the generation of mouse models for RPS19-deficient DBA using transgenic RNA interference that allows an inducible and graded down-regulation of Rps19. Rps19-deficient mice develop a macrocytic anemia together with leukocytopenia and variable platelet count that with time leads to the exhaustion of hematopoietic stem cells and bone marrow failure. Both RPS19 gene transfer and the loss of p53 rescue the DBA phenotype implying the potential of the models for testing novel therapies. This study demonstrates the feasibility of transgenic RNA interference to generate mouse models for human diseases caused by haploinsufficient expression of a gene.


Facile and statistical optimization of transfection conditions for secretion of foreign proteins from insect Drosophila S2 cells using green fluorescent protein reporter.
Insect Drosophila melanogaster S2 cells were developed as a plasmid-based and therefore nonlytic expression system for functional foreign proteins. Transfection is an important step to introduce foreign target DNA into cells and should be properly optimized to obtain maximum production yield. Single factor search (SFS) methodology is still generally used to determine optimal condition in a biological system. Although this method is relatively simple to perform, it has many disadvantages such as not considering interactions between several factors and not covering the entire region of the solution pool. Therefore, we approached this optimization problem statistically with response surface (RSM) and evolutionary operation (EVOP) methodologies and compared the transfection efficiencies with the traditional SFS method. We employed secreted green fluorescent protein (GFP) as a reporter for determination of optimal transfection condition and secreted human erythropoietin (hEPO) as a confirming foreign model protein. Consequently, we arrived at the best optimal transient transfection condition (1 microg of plasmid DNA, 5 microg of lipofectin, 2 x 10(6) cells of initial cell number, and 18 h of transfection duration time) through a systematic access in a series of SFS, RSM, and EVOP. The secreted hEPO yield using optimal transient transfection condition by EVOP methodology was enhanced by about 1.8-fold compared to that of traditional SFS. This optimized transient transfection condition can be used as a basis for optimal stable transfections. A linear relationship between secreted GFP fluorescence intensity and secreted hEPO concentration indicated that facile and noninvasive determination of optimal transfection conditions for expression and secretion of foreign proteins in S2 cell cultures was made possible by simple measurement of GFP fluorescence.


Measurement of urinary calcium using AT89C51RD2 microcontroller.
A simple and inexpensive absorption technique for determination of calcium ion in urine samples is developed, comprising a light emitting diode (650 nm) as the light source and photodiode as the detector with AT89C51RD2 microcontroller. The design of the system and details of interface, calibration, and procedure of operation are explained in this paper. Software is developed to monitor sample processing and to display the results in liquid crystal display screen. With 15 microl sample volume, a linear output is obtained in the range of 2.5-7.5 mM calcium with a detection limit of 0.06 mM. Interferences from other cations such as monovalent ion and divalent ion are investigated in the expected range, which are normally present in clinical samples, and absorption changes over the pH range of 3-12 are also determined. This system has been demonstrated successfully for the successive assay of calcium in urine samples, with the results comparing well to those achieved and in good agreement with values obtained with the current clinical spectrophotometric method at 95% of confidence level.


[The effect of elastic taping on patients with patellofemoral syndrome].
To determine the effect of elastic taping on patients with patellofemoral pain and its impact on the onset time of vastusmedialis and vastuslateralis. Eligible patients were assigned into the treatment and control groups randomly. Patients in the treatment group received a 5-day taping with therapeutic stretch. Patients in the control group were give placebo taping without therapeutic stretch. Ultrasound therapy was applied to all of the participants as a basic treatment. The levels of pain and surface electromyography were evaluated before treatment, after the first taping and on the Gh day (without taping). Patients in the treatment group experienced immediate improvement in pain and onset time of vastusmedialis and vastuslateralis compared with the controls. On the 6th day (without taping), further improvement was achieved in pain and onset time of vastusmedialis and vastuslateralis in the treatment group compared with the controls (P < 0.05). Elastic taping can effectively improve pain and onset time of vastusmedialis and vastuslateralis.


Treatment of lepromatous ulcers using citric acid as a sole antimicrobial agent.
A prospective study was carried out to assess the role of citric acid as a sole antimicrobial agent in the management of lepromatous ulcers. Thirty-four known cases of lepromatous ulcers not responding to conventional antibiotic therapies for long duration were investigated for culture and susceptibility studies. Staphylococcus aureus (25·00%) and Klebsiella spp. (23·43%) were found to be the most common isolates. Amikacin (68·75%) and ciprofloxacin (67·18%) were found to be the most effective antimicrobial agents. Topical application of citric acid ointment resulted in complete healing in 25 (73·52%) cases. In eight cases (26·48%), there was elimination of infective agent from ulcer site and formation of healthy granulation, but no complete healing of ulcer was seen. Results indicate that citric acid is the best alternative for the effective management of lepromatous ulcers when other therapies are exhausted.


Ascertainment bias in the estimation of sibling genetic risk parameters.
The sibling recurrence risk, sibling relative risk, and locus-specific sibling relative risk are fundamental quantities in genetic epidemiologic research and are often estimated without accounting for the sampling scheme. For data generated under some genetic models, bias of estimates may be large if the sampling method is incorrectly modeled. In this paper, we explore the relationship between ascertainment of sibships and estimation and interpretation of genetic risk parameters. In particular, we observe that, although traditional definitions of these population parameters are consistent with each other, implied assumptions about ascertainment and the nature of ascertainment correction differ. In the absence of ascertainment correction, unbiased estimation of sibling recurrence risk and overall sibling relative risk requires single ascertainment, while unbiased estimation of locus-specific sibling relative risk requires complete ascertainment.


Rheo-small-angle neutron scattering at the National Institute of Standards and Technology Center for Neutron Research.
We describe the design and operation of a modified commercial rheometer to simultaneously perform rheological measurements and structural studies by small angle neutron scattering (SANS). The apparatus uses a Couette geometry shear cell allowing two of the three scattering planes to be observed by performing experiments in either the radial or tangential geometries. The device enables small angle neutron scattering patterns to be obtained simultaneously with a wide variety of rheological measurements such as stress/strain flow curves, oscillatory deformations, and creep, recovery and relaxation tests, from -20 °C to 150 °C, for samples with viscosities varying by several orders of magnitude. We give a brief report of recent experiments performed on a dispersion of acicular nanoparticles and biopolymer network under stress demonstrating the utility of such measurements. This device has been developed at the National Institute of Standards and Technology's Center for Neutron Research (NCNR) and made available to the complex fluids community as part of the standard sample environment equipment.