The DNA end-binding protein Ku regulates silencing at the internal HML and HMR loci in Saccharomyces cerevisiae.
Heterochromatin resides near yeast telomeres and at the cryptic mating-type loci, HML and HMR, where it silences transcription of the alpha- and a-mating-type genes, respectively. Ku is a conserved DNA end-binding protein that binds telomeres and regulates silencing in yeast. The role of Ku in silencing is thought to be limited to telomeric silencing. Here, we tested whether Ku contributes to silencing at HML or HMR. Mutant analysis revealed that yKu70 and Sir1 act collectively to silence the mating-type genes at HML and HMR. In addition, loss of yKu70 function leads to expression of different reporter genes inserted at HMR. Quantitative chromatin-immunoprecipitation experiments revealed that yKu70 binds to HML and HMR and that binding of Ku to these internal loci is dependent on Sir4. The interaction between yKu70 and Sir4 was characterized further and found to be dependent on Sir2 but not on Sir1, Sir3, or yKu80. These observations reveal that, in addition to its ability to bind telomeric DNA ends and aid in the silencing of genes at telomeres, Ku binds to internal silent loci via protein-protein interactions and contributes to the efficient silencing of these loci.


Body art education: the earlier, the better.
Using the major format identified for successful prevention and intervention work, five "W" questions: what, why, whom, where, and who, plus one: how, this article first summarizes the current body art literature. Then, educational strategies specifically regarding the topic of tattoos and body piercing are presented for school nurses. These strategies can be used to create applicable, realistic information for young students before they are faced with decisions related to body art. The school nurse might be the first and only credible source of information the students receive related to body art in an education/health environment. Curriculum emphasis is on the decision making leading up to the action, and the major purposes, rather than on the markings or jewelry associated with procurement. Reducing decisional conflict while promoting positive development in the areas of empowerment, self-esteem, and maturing can help achieve stronger informed decision making.


Development of highly polymorphic SNP markers from the complexity reduced portion of maize [Zea mays L.] genome for use in marker-assisted breeding.
The duplicated and the highly repetitive nature of the maize genome has historically impeded the development of true single nucleotide polymorphism (SNP) markers in this crop. Recent advances in genome complexity reduction methods coupled with sequencing-by-synthesis technologies permit the implementation of efficient genome-wide SNP discovery in maize. In this study, we have applied Complexity Reduction of Polymorphic Sequences technology (Keygene N.V., Wageningen, The Netherlands) for the identification of informative SNPs between two genetically distinct maize inbred lines of North and South American origins. This approach resulted in the discovery of 1,123 putative SNPs representing low and single copy loci. In silico and experimental (Illumina GoldenGate (GG) assay) validation of putative SNPs resulted in mapping of 604 markers, out of which 188 SNPs represented 43 haplotype blocks distributed across all ten chromosomes. We have determined and clearly stated a specific combination of stringent criteria (>0.3 minor allele frequency, >0.8 GenTrainScore and >0.5 Chi_test100 score) necessary for the identification of highly polymorphic and genetically stable SNP markers. Due to these criteria, we identified a subset of 120 high-quality SNP markers to leverage in GG assay-based marker-assisted selection projects. A total of 32 high-quality SNPs represented 21 haplotypes out of 43 identified in this study. The information on the selection criteria of highly polymorphic SNPs in a complex genome such as maize and the public availability of these SNP assays will be of great value for the maize molecular genetics and breeding community.


Decline of tuberculosis notification rate in different populations and regions in Portugal, 2010-2017.
Tuberculosis (TB) incidence declined in Portugal in recent decades, but trends differ between regions and population subgroups. We investigated these differences to inform prevention and control programmes. We extracted TB notifications from the Portuguese National TB Surveillance System (SVIG-TB) in 2010-2017, disaggregated by region, age group, nationality and HIV status. We calculated notification rates using denominators from the Portuguese National Institute of Statistics and the Joint United Nations Programme on HIV/AIDS and performed stratified time series analysis. We estimated interannual decline percentages and 95% confidence intervals (CI) using Poisson and binomial negative regression models. The overall TB notification rate decreased from 25.7 to 17.5/100,000 population from 2010 to 2017 (5.2%/year) in Portugal. Interannual decline did not differ significantly between regions, but it was smaller amongst non-Portuguese nationals (-1.57% [CI: -4.79%, 1.75%] vs -5.85% [CI: -6.98%, -4.70%] in Portuguese nationals); children under five years of age (+1.77% [CI: -4.61%, 8.58%] vs -5.38% [CI: -6.33%, -4.42%] in other age groups); and HIV-negative people (-6.47% [CI: -9.10%, -3.77%] vs -11.29% [CI; -17.51%, -4.60%] in HIV-positive). The decline in TB notification rates in Portugal during the study period has been steady. However, the decline amongst non-Portuguese nationals, children under five years of age and non-infected-HIV patients was lower. No significant differences were observed between regions. Changes in TB epidemiology in specific risk groups and geographical areas should be closely monitored to achieve the objectives of the End TB Strategy. We recommend intensifying screening of TB in the subpopulations identified.


Effect of omeprazole on bioavailability of an oral extended-release formulation of ciprofloxacin.
The effect of omeprazole on the oral bioavailability and urinary exposure of the Depomed formulation of extended-release (ER) ciprofloxacin was studied. A two-way crossover study was conducted in healthy subjects. Subjects received either a single dose of ER ciprofloxacin 1000 mg or a single dose of ER ciprofloxacin 1000 mg following three days of treatment with omeprazole 40 mg. Blood and urine samples were collected over 36 hours, and ciprofloxacin concentrations were determined using high-performance liquid chromatography. Twenty-seven subjects (16 men, 11 women) received both treatments. The mean maximum concentration, mean area under the plasma-versus-concentration curve, and mean amount of ciprofloxacin excreted in urine were similar between the two treatments and met strict bioequivalence criteria. Omeprazole did not affect the plasma or urinary pharmacokinetics of an oral ER formulation of ciprofloxacin.


Leptin and TNF-alpha promoter methylation levels measured by MSP could predict the response to a low-calorie diet.
Obesity-associated adipose tissue enlargement is characterized by an enhanced proinflammatory status and an elevated secretion of adipokines such as leptin and cytokines such as tumor necrosis factor (TNF)-alpha. Among the different mechanisms that could underlie the interindividual differences in obesity, epigenetic regulation of gene expression has emerged as a potentially important determinant. Therefore, 27 obese women (age, 32-50 years; baseline body mass index, 34.4 ± 4.2 kg/m(2)) were prescribed an 8-week low-calorie diet, and epigenetic marks were assessed. Baseline and endpoint anthropometric parameters were measured, and blood samples were drawn. Genomic DNA and RNA from adipose tissue biopsies were isolated before and after the dietary intervention. Leptin and TNF-alpha promoter methylation were measured by MSP after bisulfite treatment, and gene expression was also analyzed. Obese women with a successful weight loss (≥5% of initial body weight, n=21) improved the lipid profile and fat mass percentage (-12%, p<0.05). Both systolic (-5%, p<0.05) and diastolic (-8%, p<0.01) blood pressures significantly decreased. At baseline, women with better response to the dietary intervention showed lower promoter methylation levels of leptin (-47%, p<0.05) and TNF-alpha (-39%, p=0.071) than the non-responder group (n=6), while no differences were found between responder and non-responder group in leptin and TNF-alpha gene expression analysis. These data suggest that leptin and TNF-alpha methylation levels could be used as epigenetic biomarkers concerning the response to a low-calorie diet. Indeed, methylation profile could help to predict the susceptibility to weight loss as well as some comorbidities such as hypertension or type 2 diabetes.


The light-induced increase of carbohydrate metabolism in glial cells of the honeybee retina is not mediated by K+ movement nor by cAMP.
The retina of the honeybee drone is a nervous tissue in which glial cells and photoreceptor neurons constitute two distinct metabolic compartments. The phosphorylation of glucose and its subsequent incorporation into glycogen occur essentially in glia, whereas O2 consumption occurs in the photoreceptors. After [3H] glucose loading of superfused retinal slices, light stimulation induced a significant rise in [3H] glycogen turnover in the glia. This occurs without a concomitant covalent modification of glycogen enzymes. Probably only an increase or a decrease of the availability of [3H] glycosyls that are incorporated into glycogen is necessary. As only photoreceptors are directly excitable by light, we searched for a signal that stimulates glycogen metabolism in the glia. Although K+ in extracellular space and glia increases after repetitive light stimulation, increasing bath K+ in the dark did not mimic the metabolic effects of light, despite an equivalent increase of K+ in the extracellular space and glia. We subsequently explored the role of cAMP, a universal intracellular second messenger. Exposure of retinal slices to the adenylate-cyclase activator forskolin induced an expected increase in the rate of formation of cAMP, but only partially mimicked the metabolic effects of light. Furthermore, light stimulation failed to induce a rise in the rate of formation of cAMP. We conclude that in this nervous system, without synapses, neither K+ nor cAMP mediates the effect of light stimulation on intraglial glucose metabolism.


Fever management in children.
Treating a child with a fever is a skilled nursing activity. This article explains the underlying physiological mechanisms of a fever and relates these to the clinical signs, symptoms and care of children.


Rivaroxaban reversal with prothrombin complex concentrate or tranexamic acid in healthy volunteers.
Essentials Specific reversal agents for managing severe factor Xa inhibitor-associated bleeding are lacking. We assessed 4-factor-prothrombin complex concentrate (4F-PCC) and tranexamic acid (TXA). 4F-PCC, but not TXA, reduced the prothrombin time and increased endogenous thrombin potential. These agents may be viable options for reversal of therapeutic doses of rivaroxaban. Background Oral activated factor X inhibitors such as rivaroxaban are widely used, but specific reversal agents are lacking. Although four-factor prothrombin complex concentrate (4F-PCC) and tranexamic acid (TXA) are sometimes used to manage serious bleeding, their efficacy is unknown. Prior studies in healthy subjects taking rivaroxaban revealed that 4F-PCC partially reverses the prolonged prothrombin time (PT), and fully restores the endogenous thrombin potential (ETP). The effect of TXA has not been evaluated. Methods In this double-blind, parallel-group study, 147 healthy volunteers given rivaroxaban 20 mg twice daily for 3 days were randomized after their morning dose on day 4 to receive intravenous 4F-PCC (50 IU kg-1 ), TXA (1.0 g), or saline. Standardized punch biopsies were performed at baseline and after 4F-PCC, TXA or saline administration. Reversal was assessed by measuring bleeding duration and bleeding volume at biopsy sites, and by determining the PT and ETP. Results As compared with saline, 4F-PCC partially reversed the PT and completely reversed the ETP, whereas TXA had no effect. Neither 4F-PCC nor TXA reduced bleeding duration or volume. All treatments were well tolerated, with no recorded adverse events. Conclusions Although 4F-PCC reduced the PT and increased the ETP in volunteers given supratherapeutic doses of rivaroxaban, neither 4F-PCC nor TXA influenced punch biopsy bleeding.


Dickkopf1: An Immunomodulator in Tissue Injury, Inflammation, and Repair.
Upon injury, inflammation and repair processes are orchestrated to maintain tissue homeostasis. The Wnt ligands play essential roles in cell differentiation and proliferation for tissue repair and regeneration. It is increasingly clear that Wnt ligands play crucial immune-modulatory roles in inflammatory diseases. It is predicted that comprehensive research regarding the cross-talk between nonimmune and immune cells in tissue injury and repair will flourish. The Wnt system and immune system interaction will be critical to understanding tissue injury, inflammation, and repair. In this study, we will first introduce the Wnt system and review the role of the Wnt system in tissue regeneration and repair. We will review the previous literature regarding how the Wnt ligands regulate the immune system. Next, we will discuss the current and future perspectives of Wnt ligands to target cancer and other immunological diseases. Finally, we will discuss the quintessential Wnt antagonist Dickkopf1 as an immunomodulatory ligand.


[Connective tissue growth factor promotes the proliferation of myofibroblast through Erk-1/2 signaling pathway].
To investigate the mechanism by which connective tissue growth factor (CTGF) enhances transforming growth factor-beta(1) (TGF-beta(1))-mediated myofibroblastic activation in renal interstitial fibroblast NRK-49F. NRK-49F cells were pretreated by TGF-beta(1) so that some cells transform into myofibroblasts, and then the cells were devided into vechile, CTGF treated group, TGF-beta(1) treated group, and PD98059 intervene group. The hallmark of myofibroblast, alpha-SMA immunostaining and marker of cellular proliferation, BrdU incorporation were determined by immunocytochemistry doublestaining. The protein level of alpha-SMA was determined by Western blot analysis. CTGF induced a proliferative response in myofibroblast initiated by TGF-beta(1), whereas TGF-beta(1) had no action on proliferation. Although CTGF could not induce myofibroblastic activation in renal interstitial fibroblast, it upregulated the protein level of alpha-smooth muscle actin significantly in cells pretreated by TGF-beta1 (P < 0.05). Significant phosphorylation of Erk-1/2 was detected after incubation with CTGF for 30 min in the cells pretreated by TGF-beta(1), while TGF-beta(1) did not have this ability. Inhibition of Erk-1/2 activation by Mek kinase inhibitor PD98059 suppressed CTGF-mediated myofibroblasts proliferation and significantly down-regulated expression of alpha-SMA protein in cells pretreated by TGF-beta(1) (P < 0.05). CTGF induced a proliferative response in TGF-beta(1)-initiated myofibroblasts, and this action is likely dependent on the activation of Erk-1/2 signaling pathway.


Pathogens associated with persistent diarrhoea in children in low and middle income countries: systematic review.
Persistent diarrhoea in children is a common problem in low and middle income countries. To help target appropriate treatment for specific pathogens in the absence of diagnostic tests, we systematically reviewed pathogens most commonly associated with persistent diarrhoea in children. We sought all descriptive studies of pathogens in the stool of children with diarrhoea of over 14 days duration in low and middle income countries with a comprehensive search of the MEDLINE, EMBASE, LILACS and WEB OF SCIENCE databases. We described the study designs and populations, assessed the quality of the laboratory tests, and extracted and summarised data on pathogens. For Escherichia coli, we calculated high and low prevalence estimates of all enteropathic types combined. Results across studies were compared for geographical patterns. Nineteen studies were included. Some used episodes of diarrhoea as the unit of analysis, others used children. The quality of reporting of laboratory procedures varied, and pathogens (particularly E. coli types) were classified in different ways. As there were no apparent regional differences in pathogen prevalence, we aggregated data between studies to give a guide to overall prevalence. Enteropathic E. coli types were commonly found in children with persistent diarrhoea (up to 63%). Various other organisms, including viruses, bacteria and parasites, were detected but across all studies their prevalence was under 10%. However, these pathogens were also found in similar frequencies in children without diarrhoea. A number of pathogens are commonly associated with persistent diarrhoea in children, but in children without diarrhoea the pathogens are found with similar frequencies. New research with carefully selected controls and standardised laboratory investigations across countries will help map causes and help explore effective options for presumptive treatment.


A Unique Model for ONSD Part II: Inter/Intra-operator Variability.
To evauluate our novel ultrasound model for measurement of optic nerve sheath diameter (ONSD) and determine the intra- and inter-operator variability associated with this technique. We conducted ten measurements of ONSD per model amongst eight different models with a single experienced operator to examine intra-operator variability. Similarly, we had seven different operators measure the OSND twice in eight different models, in order to determine inter-operator variability analyzed with a three level linear statistical model. For intra-operator variability, the intra-cluster correlation coefficients for the experienced and novice operators were 0.643 and 0.453 respectively. This displayed improvement in intra-operator variability with experience. The inter-cluster correlation coefficient was 0 for the group of novice operators, indicating negligible difference amongst multiple operators in measuring any given model of ONSD. A strong, statistically significant, linear relationship between the actual model disc size and the ultrasound ONSD measures was identified, implying the reliability of the images produced by our novel model. Utilizing a novel model for ONSD ultrasonography, we have determined the intraoperator reliability of ONSD measurement to be moderate, with no appreciable difference amongst multiple operators. Improvement in measurement reliability has been demonstrated between expert and novice operators with our model, indicating the potential benefit of simulation platforms for teaching the technique of ONSD ultrasound.


Effect of chlordiazepoxide on schedule-controlled responding and schedule-induced drinking.
Lever pressing of four rats was maintained under a multiple fixed ratio 80, fixed interval 2-min schedule of food presentation. Water was concurrently available from a drinking tube. Overall rates of lever pressing were highest under the fixed ratio schedule and, for three rats, most drinking occurred during the pause preceding responding under the fixed interval schedule. Chlordiazepoxide increased the lower rates of lever pressing maintained under the fixed interval schedule but generally decreased the higher response rates under the fixed ratio schedule. The effects of chlordiazepoxide on schedule-induced licking also depended on the extent to which this response occurred in each schedule component. Typically chlordiazepoxide produced relatively greater increases in the lower levels of licking and either increased less or decreased licking in that component where, under control conditions, this response was more extensive. Chlordiazepoxide also produced overall increases in the total amount of water consumed during the session. When the number of food pellets obtained during the experimental session was given all at one time in the home cage, the amount of water ingested over a period of time equivalent to the session duration was substantially less than that consumed during the experimental session. Chlordiazepoxide did not increase home cage water consumption under this condition.


Pharmacological evaluation of JO 1870: relation to the potential treatment of urinary bladder incontinence.
The effect of p.o. administered JO 1870 [(-)-1-(p-chlorophenyl)-N,N- dimethyl-1-ethyl(4-phenyl)-but-3-en-1-ylamine, hydrochloride] on the urinary bladder cystometrogram has been investigated in the unanesthetized rat. The effects of JO 1870 have been compared with those of morphine and reference drugs currently used for the treatment of urinary incontinence in humans. JO 1870 (2.5-25 mg/kg) dose-dependently increased the bladder capacity and the threshold pressure responsible for urination. These effects were antagonized by the i.v. administration of naloxone (0.2 mg/kg). In comparison, morphine (2.5-25 mg/kg) moderately increased urinary volume and threshold pressure. Different anticholinergic drugs (propantheline bromide, 10 mg/kg; terodiline, 25 mg/kg; dicyclomine, 25 mg/kg) had no significant effects on either parameter and the antidepressant clomipramine (25 mg/kg) decreased both. In vitro, JO 1870 specifically displaced [3H]-[D-Ala2,MePhe4,Gly5-(ol)]enkephalin from binding sites in guinea pig whole-brain membranes and rat thalamus; the sodium shift ratio obtained from [3H]naloxone binding was 16. These results suggest that JO 1870 has some opioid agonist activity. JO 1870 (2.5-300 mg/kg) had few effects on cardiorespiratory, gastrointestinal or nociceptive systems in rats. Together, these results indicate that JO 1870 is a nonanticholinergic agent that potently increases bladder capacity, likely through an opioid mechanism, and may have potential use in the treatment of urinary incontinence.


Evidence for deep phylogenetic conservation of exonic splice-related constraints: splice-related skews at exonic ends in the brown alga Ectocarpus are common and resemble those seen in humans.
The control of RNA splicing is often modulated by exonic motifs near splice sites. Chief among these are exonic splice enhancers (ESEs). Well-described ESEs in mammals are purine rich and cause predictable skews in codon and amino acid usage toward exonic ends. Looking across species, those with relatively abundant intronic sequence are those with the more profound end of exon skews, indicative of exonization of splice site recognition. To date, the only intron-rich species that have been analyzed are mammals, precluding any conclusions about the likely ancestral condition. Here, we examine the patterns of codon and amino acid usage in the vicinity of exon-intron junctions in the brown alga Ectocarpus siliculosus, a species with abundant large introns, known SR proteins, and classical splice sites. We find that amino acids and codons preferred/avoided at both 3' and 5' ends in Ectocarpus, of which there are many, tend, on average, to also be preferred/avoided at the same exon ends in humans. Moreover, the preferences observed at the 5' ends of exons are largely the same as those at the 3' ends, a symmetry trend only previously observed in animals. We predict putative hexameric ESEs in Ectocarpus and show that these are purine rich and that there are many more of these identified as functional ESEs in humans than expected by chance. These results are consistent with deep phylogenetic conservation of SR protein binding motifs. Assuming codons preferred near boundaries are "splice optimal" codons, in Ectocarpus, unlike Drosophila, splice optimal and translationally optimal codons are not mutually exclusive. The exclusivity of translationally optimal and splice optimal codon sets is thus not universal.


The quality of impressions for crowns and bridges: an assessment of the work received at three commercial dental laboratories. assessing qualities of impressions that may lead to occlusal discrepancies with indirect restorations.
There are few published studies that directly assess the quality of impressions for crowns and bridges in the UK. This paper considers aspects of impression quality with particular attention to factors causing potential occlusal discrepancies in the final restoration. To this end three dental laboratories were visited over a 3-month period. All impressions for conventional crown and bridgework that arrived on the days of the visits were examined and assessed against criteria defined on a custom-designed assessment form. A total of 206 impression cases were considered in this study. Flexible impression trays were used for 65% of working impressions. Their use was more common for NHS work than for private work. 31.9% of all alginate impressions examined were not adequately fixed to the tray. Visible contamination of impressions was not uncommon.


The multifaceted roles of microtubule-associated proteins in the primary cilium and ciliopathies.
The primary cilium is a conserved microtubule-based organelle that is critical for transducing developmental, sensory and homeostatic signaling pathways. It comprises an axoneme with nine parallel doublet microtubules extending from the basal body, surrounded by the ciliary membrane. The axoneme exhibits remarkable stability, serving as the skeleton of the cilium in order to maintain its shape and provide tracks to ciliary trafficking complexes. Although ciliary trafficking and signaling have been exhaustively characterized over the years, less is known about the unique structural and functional complexities of the axoneme. Recent work has yielded new insights into the mechanisms by which the axoneme is built with its proper length and architecture, particularly regarding the activity of microtubule-associated proteins (MAPs). In this Review, we first summarize current knowledge about the architecture, composition and specialized compartments of the primary cilium. Next, we discuss the mechanistic underpinnings of how a functional cilium is assembled, maintained and disassembled through the regulation of its axonemal microtubules. We conclude by examining the diverse localizations and functions of ciliary MAPs for the pathobiology of ciliary diseases.


Cardiolipin prevents rate-dependent uncoupling and provides osmotic stability in yeast mitochondria.
The role of cardiolipin in mitochondrial function was studied by comparing the energy-transforming and osmotic properties of mitochondria isolated from the Saccharomyces cerevisiae cardiolipin synthase-null mutant crd1Delta, which has no cardiolipin, and the isogenic wild type. The results indicated that the importance of cardiolipin for energetic coupling strongly depends on the rate of oxidative phosphorylation, which was set by using NADH (maximal rate limited by coupling mechanism) or ethanol (moderate rate limited by electron supply) as a respiratory substrate, or by modulating the steady-state rate of NADH supply. The absence of cardiolipin resulted in only a small effect on oxidative phosphorylation proceeding at a moderate rate, but led to significant uncoupling (decreased ADP/O and increased state 4 respiration) at the maximal rate of respiration. This indicates that cardiolipin prevents rate-dependent uncoupling in the energy-transforming apparatus. This role of cardiolipin may derive from its strong interaction with, and modulation of the function of, respiratory complexes, and from its effects on the physical properties of the membrane. The importance of cardiolipin for mitochondrial osmotic properties was determined by comparing oxidative phosphorylation, release of matrix enzyme, shrinking ability and volume dynamics upon hypotonically induced swelling in crd1Delta and wild-type mitochondria. Opening of the yeast mitochondrial unspecific channel (YMUC) in the wild-type and mutant mitochondria was also tested. It was found that the lack of cardiolipin strongly undermines the osmotic stability of the mitochondrial membrane.


Randomised controlled trial of oxygen therapy and high-flow nasal therapy in African children with pneumonia.
The life-saving role of oxygen therapy in African children with severe pneumonia is not yet established. The open-label fractional-factorial COAST trial randomised eligible Ugandan and Kenyan children aged > 28 days with severe pneumonia and severe hypoxaemia stratum (SpO2 < 80%) to high-flow nasal therapy (HFNT) or low-flow oxygen (LFO: standard care) and hypoxaemia stratum (SpO2 80-91%) to HFNT or LFO (liberal strategies) or permissive hypoxaemia (ratio 1:1:2). Children with cyanotic heart disease, chronic lung disease or > 3 h receipt of oxygen were excluded. The primary endpoint was 48 h mortality; secondary endpoints included mortality or neurocognitive sequelae at 28 days. The trial was stopped early after enrolling 1852/4200 children, including 388 in the severe hypoxaemia stratum (median 7 months; median SpO2 75%) randomised to HFNT (n = 194) or LFO (n = 194) and 1454 in the hypoxaemia stratum (median 9 months; median SpO2 88%) randomised to HFNT (n = 363) vs LFO (n = 364) vs permissive hypoxaemia (n = 727). Per-protocol 15% of patients in the permissive hypoxaemia group received oxygen (when SpO2 < 80%). In the severe hypoxaemia stratum, 48-h mortality was 9.3% for HFNT vs. 13.4% for LFO groups. In the hypoxaemia stratum, 48-h mortality was 1.1% for HFNT vs. 2.5% LFO and 1.4% for permissive hypoxaemia. In the hypoxaemia stratum, adjusted odds ratio for 48-h mortality in liberal vs permissive comparison was 1.16 (0.49-2.74; p = 0.73); HFNT vs LFO comparison was 0.60 (0.33-1.06; p = 0.08). Strata-specific 28 day mortality rates were, respectively: 18.6, 23.4 and 3.3, 4.1, 3.9%. Neurocognitive sequelae were rare. Respiratory support with HFNT showing potential benefit should prompt further trials.


[Lipid peroxidation and microviscosity of erythrocyte membranes in patients with schizophrenia].
During examination of 27 patients with schizophrenia, a study was made of the processes of free radical lipid oxidation. It has been established that in patients with schizophrenia, the processes of lipid peroxidation are activated and microviscosity of erythrocytic membranes is increased. The rise of microviscosity of erythrocytic membranes is attended by an increase of cholesterol content and of the amount of sialic acids of gangliosides involved in the formation of the superficial charge of erythrocytes. The authors hold that the changes discovered may cause functional destabilization of erythrocytic membranes, which may play a definite part in the development of hypoxia in schizophrenic patients.


NSAIDs associated with increased risk of congestive heart failure in elderly patients taking diuretics.
Both diuretics and nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used, in particular among the elderly. The use of NSAIDs may decrease the efficacy of diuretics and induce congestive heart failure (CHF) in patients treated with diuretics. To investigate the risk of CHF associated with combined use of diuretics and NSAIDs in patients older than 55 years. We conducted a study in a base cohort of 10,519 recipients of diuretics and NSAIDs identified in the PHARMO database during the period from 1986 through 1992. The incidence density of hospitalizations for CHF during exposure to both diuretics and NSAIDs (index) was compared with that during exposure to diuretics only (reference). We found an overall increased risk of hospitalization for CHF during periods of concomitant use of diuretics and NSAIDs compared with use of diuretics only (crude relative risk, 2.2; 95% confidence interval, 1.7-2.9). After adjusting for cofactors including age, sex, history of hospitalization, and drug use, a 2-fold increased risk remained (relative risk, 1.8; 95% confidence interval, 1.4-2.4). Use of NSAIDs in elderly patients taking diuretics is associated with a 2-fold increased risk of hospitalization for CHF, especially in those with existing serious CHF.


RH maps of bovine chromosomes 15 and 29: conservation of human chromosomes 11 and 5.
Comparative mapping data on evolutionary conserved coding sequences and synteny maps between human and cattle are insufficient to define the extent and distribution of conserved segments between these two species, because the order of loci is often rearranged. A 5000-rad cattle whole-genome radiation hybrid (WG-RH) panel was constructed to provide high-resolution comparative maps and also to integrate linkage maps of microsatellites with evolutionary conserved genes and transcripts in a single ordered map. We used the WG-RH panel to construct radiation hybrid maps of bovine Chromosomes (Chrs) 15 and 29 (BTA15 and BTA29), integrating microsatellites from published linkage maps with selected genes. The comprehensive map of BTA15 consists of 24 markers, 13 of which were placed in the framework map. Eleven molecular markers compose the comprehensive map of BTA29, seven of which were placed in the framework map. We identified the homologous regions between bovine Chr 15 (BTA15) and human Chrs 5 and 11 (HSA5 and HSA11), as well as between BTA29 and HSA11. The present study demonstrates that WG-RH mapping is an efficient method for integrating multiple genetic maps into one map and for incorporating monomorphic Type I loci into ordered maps for comparison between species.


Single-frequency, thin-film-tuned, 0.6-W, diode-pumped Nd:YVO(4) laser.
Application of a metallic thin-film selector to the single-frequency oscillation of a diode-pumped Nd:YVO(4) laser has been investigated theoretically and experimentally. We show that a chromium thin-film selector with a thickness between 8 and 9 nm provides single-frequency output within a power range of 0.6 W. Single-frequency operation, slow smooth tuning, or chirping was realized by the output coupler movement with a piezoceramic transducer. Chirping at a repetition rate of 0.5 kHz in the 0.5-10-GHz range was achieved. Physical and technical limitations caused by the wide-gain bandwidth, thermal effects, and mechanical vibrations of cavity elements are discussed.


The diversity and breadth of cancer cell fatty acid metabolism.
Tumor cellular metabolism exhibits distinguishing features that collectively enhance biomass synthesis while maintaining redox balance and cellular homeostasis. These attributes reflect the complex interactions between cell-intrinsic factors such as genomic-transcriptomic regulation and cell-extrinsic influences, including growth factor and nutrient availability. Alongside glucose and amino acid metabolism, fatty acid metabolism supports tumorigenesis and disease progression through a range of processes including membrane biosynthesis, energy storage and production, and generation of signaling intermediates. Here, we highlight the complexity of cellular fatty acid metabolism in cancer, the various inputs and outputs of the intracellular free fatty acid pool, and the numerous ways that these pathways influence disease behavior.


Growth and characterization of LNCaP prostate cancer cell spheroids.
Cells from the prostate tumor cell line LNCaP have been grown as spheroids. The growth kinetics of the spheroids have been characterized by fitting a Gompertz equation to spheroid growth curves. The proliferation state of cells within spheroids of different diameters was assessed by bromodeoxyuridine staining. Scanning and electron transmission microscopy were performed to determine the ultrastructure of the spheroids. Prostate-specific antigen (PSA) secretion was monitored throughout spheroid growth. Consistent with Gompertzian kinetics, the volume of LNCaP spheroids initially increased exponentially and then reached a plateau. The doubling time during the exponential phase was 29 +/- 4 h. A core of nonproliferating cells was seen in spheroids with a diameter of 400 microm; at a diameter of 600 microm, a necrotic core had formed. In smaller, 200-microm diameter spheroids, a core of nonproliferating cells was not seen, but proliferating cells were concentrated at the spheroid periphery. Electron microscopy showed that the spheroids were enveloped by an extracellular matrix and that cell adhesion within the spheroids was due in part to desmosomes. PSA secretion by the spheroids could be modeled as originating from a spherical shell whose thickness was independent of overall spheroid diameter. The shell thickness obtained by fitting an appropriate equation to the data was consistent with that determined from the bromodeoxyuridine studies. LNCaP cells exhibit several important features of prostate cancer cells; in vivo, they are androgen responsive, and they express prostatic acid phosphatase, PSA, and prostate-specific membrane antigen. LNCaP spheroids provide a simple but relevant model for the study of drug delivery and response in prostate cancer.


The extraction and purification of a cysteine transfer ribonucleic acid from baker's yeast.
1. A modification of the RPC 1 system of A.D. Kelmers, G.D. Novelli & M.P. Stulberg (1965) (J. Biol. Chem. 240, 3979-3983) is described in which the support medium is a Celite of narrow range particle size treated with dichlorodimethylsilane. 2. By using this system an apparently pure preparation of tRNA Cys was isolated from baker's yeast tRNA. 3. This preparation accepted at least 60% of the theoretical quantity of [3-14C]cysteine in a conventional assay and failed to accept isoleucine, phenylalanine, proline, serine or tyrosine. 4. A theoretical countercurrent-distribution curve calculated by assuming a distribution coefficient K of 2.03 was in excellent agreement with the profiles of E260 and cysteine-acceptor ability after 537 transfers in the 1.85 M-phosphate/formamide/propan-2-ol system of C.M. Connelly & B.P. Doctor (1965) (J. Biol. Chem. 241, 715-719). 5. Chromatography of tRNA Cys on Bio-Gel P100 polyacrylamide beads afforded two components one of which was far less efficient than the other in accepting cysteine. The base compositions of the two were similar.


Basolateral potassium channel in turtle colon. Evidence for single-file ion flow.
Treatment of the apical surface of the isolated, ouabain-inhibited turtle colon with the polyene antibiotic amphotericin B permitted the properties of a barium-sensitive potassium conductance in the basolateral membrane to be discerned from the measurements of transepithelial fluxes and electrical currents. Simultaneous measurements of potassium currents and 42K fluxes showed that the movement of potassium was not in accord with simple diffusion. Two other cations, thallium and rubidium, were also permeable and, in addition, exhibited strong interactions with the potassium tracer fluxes. The results indicate that permeant cations exhibit positive coupling, which is consistent with a single-file mechanism of ion translocation through a membrane channel.


Trichoderma species associated with the green mold epidemic of commercially grown Agaricus bisporus.
Trichoderma aggressivum sp. nov. and T. aggressivum f. europaeum f. nov. are described. These forms cause the green mold epidemic in commercially grown Agaricus bisporus in North America and Europe, respectively. In the literature they have been reported as T. harzianum biotypes Th 4 and Th 2, respectively. They are strongly separated from their closest relative, T. harzianum, in sequences of the ITS-1 region of nuclear rDNA and an approximately 689 bp fragment of the protein coding translation elongation factor gene (EF-1α). They are distinguished from the morphologically similar T. harzianum and T. atroviride (the latter also known as biotype Th 3) most readily by rate of growth. Of these, only T. harzianum grows well and sporulates at 35 C, while T. atroviride is the slowest growing. Trichoderma aggressivum f. aggressivum and f. europaeum are effectively indistinguishable morphologically although they have subtly different growth rates at 25 C on SNA and statistically significant micromorphological differences. Based on findings of this study, descriptions of T. harzianum and T. atroviride are expanded. A key to Trichoderma species commonly found associated with commercially grown A. bisporus is provided.


Requirements for catalysis in mammalian glycogenin.
Glycogenin is a glycosyltransferase that functions as the autocatalytic initiator for the synthesis of glycogen in eukaryotic organisms. Prior structural work identified the determinants responsible for the recognition and binding of UDP-glucose and the catalytic manganese ion and implicated two aspartic acid residues in the reaction mechanism for self-glucosylation. We examined the effects of substituting asparagine and serine for the aspartic acid residues at positions 159 and 162. We also examined whether the truncation of the protein at residue 270 (delta270) was compatible with its structural integrity and its functional role as the initiator for glycogen synthesis. The truncated form of the enzyme was indistinguishable from the wild-type enzyme by all measures of activity and could support glycogen accumulation in a glycogenin-deficient yeast strain. Substitution of aspartate 159 by either serine or asparagine eliminated self-glucosylation and reduced trans-glucosylation activity by at least 260-fold but only reduced UDP-glucose hydrolytic activity by 4-14-fold. Substitution of aspartate 162 by either serine or asparagine eliminated self-glucosylation activity and reduced UDP-glucose hydrolytic activity by at least 190-fold. The trans-glucosylation of maltose was reduced to undetectable levels in the asparagine 162 mutant, whereas the serine 162 enzyme showed only an 18-30-fold reduction in its ability to trans-glucosylate maltose. These data support a role for aspartate 162 in the chemical step for the glucosyltransferase reaction and a role for aspartate 159 in binding and activating the acceptor molecule.


Myocilin levels in primary open-angle glaucoma and pseudoexfoliation glaucoma human aqueous humor.
To determine the concentration of myocilin in primary open-angle glaucoma (POAG) and pseudoexfoliation glaucoma (PEXG) aqueous humor. Aqueous humor was collected during surgery from patients with POAG, PEXG, and elective cataract removal (control). Volume-equivalent aqueous samples were separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gradient gels. Quantification of myocilin levels was performed using Western blots probed with 2 independent N-terminal polyclonal anti-myocilin antibodies (AB1 and AB2) followed by densitometry. Myocilin levels in aqueous humor were quantified by plotting the densitometry readings of the aqueous samples against a recombinant myocilin standard curve. Total protein concentration was determined by Bradford protein assay. Transforming growth factor β 2 levels were assessed by enzyme-linked immunosorbent assay. Myocilin levels are significantly elevated in human POAG aqueous humor when compared with control aqueous humor (AB1: 0.66±0.53 ng/μL vs. 0.23±0.20 ng/μL, P<0.001; AB2: 0.98±0.59 ng/μL vs. 0.65±0.5 ng/μL, P<0.03; mean±SD). Myocilin makes up a larger percent of the total protein in POAG aqueous humor compared with control aqueous (AB1: 0.26±0.20% vs. 0.10±0.20%, P<0.001; AB2: 0.43±0.32% vs. 0.28±0.18%, P<0.05). In contrast to POAG, myocilin levels were not elevated in PEXG aqueous humor when compared with control aqueous humor. No correlation between myocilin and transforming growth factor β 2 levels was observed. Myocilin is elevated in POAG, but not in PEXG aqueous humor.


Cross-sectional study of the prevalence of and risk factors for hoof disorders in horses in The Netherlands.
Information is scarce on the prevalence of hoof disorders in horses. In this study, we examined the prevalence of and risk factors for hoof disorders in a population of horses in The Netherlands. In a group of 942 randomly selected horses, hoof health was scored during regular foot trimming (one horse/farm). Hooves were assessed for the occurrence of one of 12 hoof disorders by a group of 21 certified farriers in two periods i.e. winter and summer of 2015. The mean age of the group of horses was 11.2±5.6years. They were mainly used for recreation (28.2%), dressage (26.8%), other disciplines (such as carriage driving and breeding) (18.7%), showjumping (17.6%) or combinations of these activities (8.6%). The horse farms studied were evenly distributed throughout the country. The horses were housed on different types of bedding, including straw (51.0%), shavings (17.5%), flax (16.1%) or other materials (11.0%), or were kept at pasture (4.4%). In 85% of the horses, at least one hoof disorder was observed during regular foot trimming. Most of the lesions were mild. The most frequently diagnosed hoof disorders were: thrush (T; 45.0%); superficial hoof wall cracks (SHWC; 30.4%); growth rings (GR; 26.3%); and sole bruises (SB; 24.7%). Less frequently observed hoof disorders included: perforating hoof wall cracks (PHWC; 16.4%); white line disease (WLD; 17.8%); and white line widening (WLW; 11.8%). Horizontal hoof cracks (5.2%), chronic laminitis (3.9%), quarter cracks (2.7%), keratoma (1.8%) and frog cancer (1.0%) were less frequently observed. Factors significantly associated with the occurrence of thrush comprised a wet stable floor (OR 1.6 and 2.9, for somewhat wet to wet respectively, compared to dry), the use of straw as bedding (OR=1.5, compared to flax), the housing strategy (e.g. permanent housing in contrast to permanent pasturing) (OR=1.7) and poor horn quality (OR=3.4). A higher prevalence of WLD was associated with less frequent hoof picking (OR=2.1 if performed weekly instead of daily), the use of flax bedding (OR=2.1, compared to straw) and poor horn quality (OR=8.1). A higher prevalence of SB was observed in horses used for multiple disciplines (OR=3.5, compared to dressage), with white-coloured hooves (OR=5.0, compared to black hooves), with longer intervals between trimming sessions (OR=4.8 in case of 8-10 weeks compared to weekly) and with poor horn quality (OR=5.4). A higher prevalence of WLW was observed in older horses (OR=15.5 for horses >19years, compared to <5years), in those with longer intervals between trimming sessions (OR=1.8 in case of 8-10 weeks compared to weekly), and in certain breeds (OR=3.2 for Friesian horses, 2.9 for Welsh ponies and 13.1 for Shetland ponies, all compared to Dutch Warmblood). In conclusion, although most of the hoof disorders identified were only in a mild stage, still an unexpectedly high prevalence of hoof disorders was observed during regular hoof trimming. Analysis of the data showed that some parameters, such as the use of flax bedding, may be protective for certain hoof disorders but a risk factor for others. This study provides useful guidelines for monitoring hoof health, reducing lameness and optimizing equine welfare.


Dental practice during a world cruise: characterisation of oral health at sea.
To describe oral health of passengers and crew attending the dental service aboard during a two months world cruise. In a retrospective, descriptive epidemiologic study design the routine documentation of all dental treatment provided at sea was analysed after the voyage. Subjects were n = 57 passengers (3.5 % of 1619) with a mean age of 71 (+/- 9.8) years and n =56 crew (5.6 % of 999) with a mean age of 37 (+/- 12.0) years. Age, gender, nationality, number of natural teeth and implants were extracted. The prosthetic status was described by recording the number of teeth replaced by fixed prosthesis and number of teeth replaced by removable prosthesis. Oral health-related quality of life (OHRQoL) was measured using the 14-item Oral Health Impact Profile (OHIP-14) and characterised by the OHIP sum score. Women attended for treatment more often than men. Passengers had a mean number of 20 natural teeth plus substantial fixed and removable prosthodontics. Crew had a mean of 26 teeth. British crew and Australian passengers attended the dental service above average. Crew tended to have a higher average OHIP-14 sum score than passengers indicating an increased rate of perceived problems. Emergency patients from both crew and passengers have a higher sum score than patients attending for routine treatment. In passengers the average number of teeth appears to be higher than that of an age matched population of industrialized countries. However, the passengers' socioeconomic status was higher which has an effect on this finding. Socioeconomic factors also serve to explain the high standard of prosthetic care in passengers. Crew in general present with less sophisticated prosthetic devices. This is in line with their different socioeconomic status and origin from developing countries. The level of dental fees aboard in comparison to treatment costs in home countries may explain some of the differences in attendance. Passengers have enjoyed high standards of prosthetic care in the past and will expect a similarly high standard from ship based facilities. The ease of access to quality dental care may explain the relatively low level of perceived problems as characterised by oral health-related quality of life scores. The dental officer aboard has to be prepared to care for very varied diagnostic and treatment needs.


Using egocentric analysis to investigate professional networks and productivity of graduate students and faculty in life sciences in Japan, Singapore, and Taiwan.
Prior studies showed that scientists' professional networks contribute to research productivity, but little work has examined what factors predict the formation of professional networks. This study sought to 1) examine what factors predict the formation of international ties between faculty and graduate students and 2) identify how these international ties would affect publication productivity in three East Asian countries. Face-to-face surveys and in-depth semi-structured interviews were conducted with a sample of faculty and doctoral students in life sciences at 10 research institutions in Japan, Singapore, and Taiwan. Our final sample consisted of 290 respondents (84 faculty and 206 doctoral students) and 1,435 network members. We used egocentric social network analysis to examine the structure of international ties and how they relate to research productivity. Our findings suggest that overseas graduate training can be a key factor in graduate students' development of international ties in these countries. Those with a higher proportion of international ties in their professional networks were likely to have published more papers and written more manuscripts. For faculty, international ties did not affect the number of manuscripts written or of papers published, but did correlate with an increase in publishing in top journals. The networks we examined were identified by asking study participants with whom they discuss their research. Because the relationships may not appear in explicit co-authorship networks, these networks were not officially recorded elsewhere. This study sheds light on the relationships of these invisible support networks to researcher productivity.


Tramadol prescribed use in general and chronic noncancer pain: a nationwide register-based cohort study of all patients above 16 years.
Background and aims In the Western world, it has become clear that we are facing a crisis of overuse, abuse and improperly prescribed use of opioids. As part of the ongoing discussion on opioid use, the use and prescription of tramadol have been addressed in recent years. A significant portion of this discussion should adequately address the risk factors for the use of weak opioid products such as tramadol. The risk factors which characterise the long-term tramadol use are still incompletely understood. Thus, we aimed to describe the characteristics of Danish patients using tramadol in more detail, under different scenarios and determinants of subsequent usage patterns. Methods We conducted a nationwide cohort study to identify individuals purchasing tramadol from 01/01/2004 to 31/12/2015 who are age 16 + years old by using data from The Danish National Databases; these databases consist of unique information for all citizens in Denmark. Logistic regression analyses were used to assess the potential risk factors for repeated tramadol use. Results The final cancer-free cohort consisted of N = 941,839 tramadol users: 54.4% women, with a mean age of 53.2 years. The number of chronic noncancer pain (CNCP) was 430,641 individuals, and 56% of the total third who repeated the use of tramadol with two + purchased prescriptions were CNCP patients. The increased risk of repeated use for CNCP was, among others, associated with: male sex (HR 1.21), age 69-110 (HR 1.72), back/spine pain men (HR 1.47), women (HR 1.46), spondylopathies (HR 1.24), male osteoporosis (HR 1.22), multimorbid ulcer/skin (HR 1.28), region of municipality Northern Jutland (HR 1.74), Central Jutland (HR 1.75), number of co-medication 4-9 (HR 1.33), dementia (HR 1.27). Factors associated with decreased risk: co-medication ischemic heart disease (HR 0.85), diagnosis headache (HR 0.70), household income highest tertile (HR 0.81), unknown (HR 0.70), single women (HR 0.96). Conclusions This study proved a widespread prescribed use of tramadol in Denmark, and, as know from the literature, weak opioid use may lead to long-term use of high potent opioids, this usage is inappropriate, in general, but especially for the treatment of CNCP. Implications When striving to reduce the overuse of opioids, focus on the extensive use of tramadol may be essential. The current study indicates an excessive and not appropriately prescribed use of tramadol among Danish CNCP patients. In addition to being inappropriate, such use may also have an impact on the growing problem of an illicit Internet market for this drug. Thus, the situation must be taken seriously. The current study confirms the recent clinical guideline and the National Recommendations in Denmark, which emphasises the risks of problematic use of tramadol. The research may also be relevant in other comparable countries. Caution must especially be taken with CNCP patients with comorbidities like diabetes, lung disease, dementia, and osteoporosis.


Investigation of retinoic acid function during embryonic brain development using retinaldehyde-rescued Rdh10 knockout mice.
Retinoic acid (RA) signaling controls patterning and neuronal differentiation within the hindbrain, but forebrain RA function remains controversial. RA is produced from metabolism of retinol to retinaldehyde by retinol dehydrogenase (RDH), followed by metabolism of retinaldehyde to RA by retinaldehyde dehydrogenase (RALDH). Previous studies on Raldh2-/- and Raldh3-/- mice demonstrated an RA requirement for γ-aminobutyric acid (GABA)ergic and dopaminergic differentiation in forebrain basal ganglia, but no RA requirement was observed during early forebrain patterning or subsequent forebrain cortical expansion. However, other studies suggested that RA controls forebrain patterning, and analysis of ethylnitrosourea-induced Rdh10 mutants suggested that RA synthesized in the meninges stimulates forebrain cortical expansion. We generated Rdh10-/- mouse embryos that lack RA activity early in the head and later in the meninges. We observed defects in hindbrain patterning and eye RA signaling, but early forebrain patterning was unaffected. Retinaldehyde treatment of Rdh10-/- embryos from E7-E9 rescues a cranial skeletal defect, resulting in E14.5 embryos lacking meningeal RA activity but maintaining normal forebrain shape and cortical expansion. Rdh10-/- embryos demonstrate that RA controls hindbrain but not early forebrain patterning, while studies on retinaldehyde-rescued Rdh10-/- embryos show that meningeal RA synthesis is unnecessary to stimulate forebrain cortical expansion.


Current and future trends in men's health clinics.
Men's health is a unique field that requires a multi-disciplinary approach to appropriately treat the full spectrum of men's health needs. Given that interest in men's health is relatively new, the definition of a men's health clinic continues to evolve. Originally conceived as testosterone replacement centers, men's health clinics are increasingly understood to encompass male endocrine, surgical, urologic, physical performance, and psychological issues. As interest in men's health continues to grow, it is important to focus on the field's future, including pharmacies, telemedicine, and the diffusion of healthcare.


Changing frequency of equivocal HER-2/neu scores and factors predictive of negative HER 2/neu fluorescent in situ hybridisation in invasive carcinomas of the breast.
Analysis of human epidermal growth factor receptor 2 (HER-2) status has become standard of care in breast cancer patients due to its important prognostic and therapeutic implications. Immunohistochemistry (IHC) is the most commonly used primary method for detection of HER-2 overexpression. Controversy exists on the interpretation of samples that are equivocal for HER 2 status (IHC 2+). Recent guidelines state that samples equivocal for HER 2 status require validation with fluorescent in situ hybridisation (FISH). The use of FISH, however, despite higher accuracy comes at a higher cost that is not affordable to all patients. This study is a retrospective study conducted at the American University of Beirut Medical Center, including women diagnosed with breast cancer with equivocal IHC scores presenting between 2009 and 2011. We attempted to correlate clinicopathological characteristics of patients diagnosed with breast cancer that can influence conclusions made on HER 2 status when analysing IHC equivocal samples in an effort to decrease the need for FISH testing. 113 patients in our records were included; charts were reviewed for different patient clinical characteristics and samples were analysed for pathological characteristics. Using logistic regression, progesterone receptor status and HER-2 staining of the normal glands around the tumour by IHC were the two statistically significant variables that showed association with FISH results. The strength of progesterone receptor status positivity and HER-2 staining of the normal glands around the tumour were proportional to the likelihood of a negative FISH. Also, the presence of strong and diffuse hormone receptor positivity in low-grade tumours was predictive of negative HER-2 status. In countries where resources are strained, oncologists need to think of measures to minimise the increasing financial burden of cancer care. Our study serves to highlight a few clinicopathological characteristics that might eliminate the need for further testing through FISH.


Familial visceral myopathy with pseudo-obstruction, megaduodenum, Barrett's esophagus, and cardiac abnormalities.
This report describes a new subgroup of familial visceral myopathy. Three patients from within this family were admitted to the hospital with pseudo-obstruction. Barium x-ray, abdominal plain film, esophageal manometry, colonoscopy, gastroscopy, and echocardiography were performed in all siblings for diagnostic evaluation. Two of our patients had surgery because of suspicion of acute abdomen. In one of them, full-thickness biopsy, which was performed during laparotomy, revealed findings that were compatible with familial visceral myopathy. Three siblings from this family with visceral myopathy, in which the parents were consanguineous, had megaduodenum, long-segment Barrett's esophagus, and different cardiac abnormalities.


Oxford Classification of IgA nephropathy 2016: an update from the IgA Nephropathy Classification Working Group.
Since the Oxford Classification of IgA nephropathy (IgAN) was published in 2009, MEST scores have been increasingly used in clinical practice. Further retrospective cohort studies have confirmed that in biopsy specimens with a minimum of 8 glomeruli, mesangial hypercellularity (M), segmental sclerosis (S), and interstitial fibrosis/tubular atrophy (T) lesions predict clinical outcome. In a larger, more broadly based cohort than in the original Oxford study, crescents (C) are predictive of outcome, and we now recommend that C be added to the MEST score, and biopsy reporting should provide a MEST-C score. Inconsistencies in the reporting of M and endocapillary cellularity (E) lesions have been reported, so a web-based educational tool to assist pathologists has been developed. A large study showed E lesions are predictive of outcome in children and adults, but only in those without immunosuppression. A review of S lesions suggests there may be clinical utility in the subclassification of segmental sclerosis, identifying those cases with evidence of podocyte damage. It has now been shown that combining the MEST score with clinical data at biopsy provides the same predictive power as monitoring clinical data for 2 years; this requires further evaluation to assess earlier effective treatment intervention. The IgAN Classification Working Group has established a well-characterized dataset from a large cohort of adults and children with IgAN that will provide a substrate for further studies to refine risk prediction and clinical utility, including the MEST-C score and other factors.


Compression mechanisms of ferroelectric PbTiO3 via high pressure neutron scattering.
Switchable atomic displacements generate electric dipole moments in ferroelectric materials utilized in many contemporary devices. Lead titanate, a perovskite oxide with formula PbTiO3, has been referred to as a textbook example of a prototype displacive ferroelectric and is a testing platform of widely used models of piezoelectric response of complex solid-solutions. PbTiO3 has been addressed by experimental and computational studies, often with apparently conflicting conclusions. To date, hydrostatic pressure experiments have been interpreted in terms of a model in which the dipole moments gradually diminish with increasing pressure until a transition to a cubic phase, characterized by a zero average dipole moment, occurs. The model unrealistically assumes an even compression of the crystal. Here we show by high-pressure neutron powder diffraction measurements that a fast and slow shrinkage of 12-oxygen cages around Pb and octahedra around Ti, respectively, takes place. A phase diagram consolidating earlier and present results is given.


Evaluation of the Modified Diet in Renal Disease equation for calculation of carboplatin dose.
Serum creatinine (SCr)-based formulas are used to estimate glomerular filtration rate (GFR) when calculating a dosage for carboplatin using the Calvert equation, but these formulas often underestimate measured GFR. The Modified Diet in Renal Disease (MDRD) equation appears to be a more accurate estimate of GFR in patients with chronic kidney disease, but this equation has not been studied extensively in patients with cancer. To determine the absolute difference between the dose of carboplatin administered (traditional SCr-based formulas used to estimate GFR) and the dose calculated using the MDRD equation to estimate GFR and compare the frequencies of thrombocytopenia, neutropenia, and dosage modifications between subjects in whom the difference in dose was 20% or more (divergent) or less than 20% (nondivergent). A retrospective analysis was conducted using data from patients who received carboplatin. Inclusion criteria were receipt of at least 2 doses of carboplatin, either as monotherapy or combination therapy, and documentation of desired area under the concentration-time curve (AUC). Patients were excluded if the baseline values needed to estimate GFR using the MDRD equation were not available. The absolute difference between the dose of carboplatin administered and that calculated using the MDRD equation was determined and, from this comparison, the subjects were divided into 2 groups (divergent vs nondivergent). The medical records of 186 adults who received more than 2 doses of carboplatin were included in the analysis. The doses were divergent in 89 (48%) patients. The mean target AUC values were 5.3 mg/mL/min and 5.1 mg/mL/min in the divergent and nondivergent groups, respectively, and most patients received cytotoxic regimens with a relatively low risk of febrile neutropenia. The frequencies of neutropenia, thrombocytopenia, and dosage modifications were similar between the 2 groups. Use of the MDRD equation to calculate the carboplatin dosage did not appear to result in a change in the frequency of myelosuppression or the need for dose modifications compared with traditional SCr-based formulas. The traditional SCr-based formulas for the calculation of carboplatin dosage should be used to estimate carboplatin dose until more data become available regarding the use of the MDRD equation in this population.


[Dynamics of IL-22-producing cells of mice infected with Schistosoma japonicum].
To study the dynamics of IL-22-producing cells and analyze the main source of IL-22 of mice infected with Schistosoma japonicum. The lymphocytes and splenocytes were prepared from lymphnodes and spleens of mice with S. japonicum infection at different stages (0 w, 3 w, 5w, 8 w, 13 w after infection). The proportions of IL-22-producing cells were determined by FACS. The proportions of IL-22-producing cells increased significantly in the lymphocytes and splenocytes from the mice infected with S. japonicum. The IL-22-producing T cells accounted 80% of the total IL-22-producing cells. The IL-22-producing CD4- T cells increased obviously in the 3rd week after S. japonicum infection. However, the proportion of the IL-22-producing CD4+ T cells began to rise since the 3rd week post-infection, and increased significantly at the 8th week after infection. In the lymphnodes and spleens of mice infected with S. japonicum, IL-22 is produced by both T cells and non-T cells, and T cells are the most important source of IL-22. The IL-22-producing CD4- T cells increase rapidly in the early acute inflammatory response, and the IL-22-producing CD4+ T cells increase significantly during the chronic inflammatory phase.


How to improve the performances of Fecal Immunological Tests (FIT): Need for standardization of the sampling and pre-analytical phases and revision of the procedures for comparison of methods.
Lack of reference materials and standard procedures, on faecal tests leads to major problems in harmonisation of methods and do not allow the comparison of outcome data. In particular the absence of standardisation of pre-analytical characteristic was noted for faecal test methods for haemoglobin since different manufacturers have developed different sampling procedures and report units. Moreover the physical characteristics of the faecal specimen and the designs of specimen collection devices do not allow analysis of samples on different systems in consequence, faecal tests cannot be compared using standard evaluation protocols. To improve the harmonization of results generated using different analytical systems and the overall performances of test on faecal materials we propose the introduction of standard procedures for sampling and pre-analytical phase and the adoption of specific procedures based on the use of artificial biological samples for comparison of methods. Harmonization of sampling devices with the use of a standard design for pickers and a standard ratio between analyte and buffer for different manufacturers represent a mandatory step in the roadmap for harmonization of clinical laboratory measurement on faecal materials and can allow a significant standardisation of results generated by different devices.The creation of specific protocols for the evaluation and comparison of analytical methods for analyse of faeces could lead to a significant improvement in the performance of methods and systems.


Predictors of high sensitivity C-reactive protein levels in patients with systemic lupus erythematosus.
Despite the increased prevalence of cardiovascular disease in patients with systemic lupus erythematosus (SLE), little is known about the role of high sensitivity C-reactive protein (hsCRP) or whether ethnicity, gender, anthropometric measures and treatment can alter hsCRP levels. We evaluated the effects of treatment and demographic, anthropometric and socio-economic variables on hsCRP levels in SLE. High sensitivity C-reactive protein levels were measured using an immunoturbidimetric assay in 610 patients from the Hopkins Lupus Cohort, who were followed-up regularly. In stepwise multiple regression analyses, body mass index (BMI) [odds ratio (OR) 1.72, 95% confidence interval (CI) 1.34-2.20, P < 0.001], African-American ethnicity (OR 1.97, 95% CI 1.22-3.19, P < 0.01), education (OR 0.60, 95% CI 0.42-0.86, P < 0.01), statin use (OR 0.38, 95% CI 0.18-0.82, P < 0.05), estrogen use (OR 3.65, 95% CI 1.19-11.22, P < 0.05), SLE Disease Activity Index score (OR 1.76, 95% CI 1.09-2.87, P < 0.05) and cumulative prednisone dose (OR 1.27, 95% CI 1.01-1.60, P < 0.05) were significant predictors of hsCRP levels. These findings suggest that hsCRP levels should be adjusted for BMI, ethnicity, education level, disease activity and medications when conducting cardiovascular risk assessment in patients with lupus.


Management of Inflammatory Bowel Disease in the Elderly Patient: Challenges and Opportunities.
The population of older patients with inflammatory bowel disease (IBD) continues to grow, partly reflecting the aging global population in general. The debilitating effects of IBD compound age-related decrements in health and functional capacity, and make the medical management of older patients with Crohn's disease and ulcerative colitis distinctly challenging to clinicians. Here, we review the recent literature describing the pharmacologic management of IBD in this population, with focus on the safety, tolerability, and efficacy of common treatment options, such as steroids, immunomodulators, tumor necrosis factor-α antagonists, and integrin antagonists; surgical interventions in older patients are also discussed. Few studies have systematically and prospectively evaluated the clinical challenges in the medical management of IBD in this patient population, leaving a limited evidence base to which clinicians can turn to for guidance. Treatment patterns may thus be suboptimal. For example, prolonged steroid use in the elderly was found to be common, causing significant morbidity from side effects in a particularly vulnerable population. Finally, within the context of a limited evidence base, we discuss common treatment scenarios to define the parameters within which physicians can individualize care for older patients with IBD. Overall, older patients with IBD are at higher risk of adverse events and less treatment responsiveness compared with younger patients, underscoring the need for future studies to fully characterize appropriate treatment courses for this population.


Fabrication of salt-hydrogel marbles and hollow-shell microcapsules by an aerosol gelation technique.
We designed a new method for preparation of liquid marbles by using hydrophilic particles. Salt-hydrogel marbles were prepared by atomising droplets of hydrogel solution in a cold air column followed by rolling of the collected hydrogel microbeads in a bed of micrometre sized salt particles. Evaporation of the water from the resulting salt marbles with a hydrogel core yielded hollow-shell salt microcapsules. The method is not limited to hydrophilic particles and could potentially be also applied to particles of other materials, such as graphite, carbon black, silica and others. The structure and morphology of the salt-hydrogel marbles were analysed by SEM and their particle size distributions were measured. We also tested the dissolution times of the dried salt marbles and compared them with those of table salt samples under the same conditions. The high accessible surface area of the shell of salt microcrystals allows a faster initial release of salt from the hollow-shell salt capsules upon their dissolution in water than from the same amount of table salt. The results suggest that such hollow-shell particles could find applications as a table salt substitute in dry food products and salt seasoning formulations with reduced salt content without the loss of saltiness.


Tactile modulation of whisking via the brainstem loop: statechart modeling and experimental validation.
Rats repeatedly sweep their facial whiskers back and forth in order to explore their environment. Such explorative whisking appears to be driven by central pattern generators (CPGs) that operate independently of direct sensory feedback. Nevertheless, whisking can be modulated by sensory feedback, and it has been hypothesized that some of this modulation already occurs within the brainstem. However, the interaction between sensory feedback and CPG activity is poorly understood. Using the visual language of statecharts, a dynamic, bottom-up computerized model of the brainstem loop of the whisking system was built in order to investigate the interaction between sensory feedback and CPG activity during whisking behavior. As a benchmark, we used a previously quantified closed-loop phenomenon of the whisking system, touched-induced pump (TIP), which is thought to be mediated by the brainstem loop. First, we showed that TIPs depend on sensory feedback, by comparing TIP occurrence in intact rats with that in rats whose sensory nerve was experimentally cut. We then inspected several possible feedback mechanisms of TIPs using our model. The model ruled out all hypothesized mechanisms but one, which adequately simulated the corresponding motion observed in the rat. Results of the simulations suggest that TIPs are generated via sensory feedback that activates extrinsic retractor muscles in the mystacial pad. The model further predicted that in addition to the touching whisker, all whiskers found on the same side of the snout should exhibit a TIP. We present experimental results that confirm the predicted movements in behaving rats, establishing the validity of the hypothesized interaction between sensory feedback and CPG activity we suggest here for the generation of TIPs in the whisking system.


Tissue-specific contributions of pneumococcal virulence factors to pathogenesis.
We assessed the ability of Streptococcus pneumoniae mutants deficient in either choline binding protein A (CbpA), pneumolysin (Pln), pyruvate oxidase (SpxB), autolysin (LytA), pneumococcal surface protein A, or neuraminidase A (NanA) to replicate in distinct anatomical sites and translocate from one site to the next. Intranasal, intratracheal, and intravenous models of disease were assessed in 4-week-old BALB/cJ mice by quantitation of bacterial titers in the relevant organs. Mice were also observed by use of real-time bioluminescent imaging (BLI). BLI allowed visualization of the bacteria in sites not tested by sampling. All mutants were created in D39 Xen7, a fully virulent derivative of capsular type 2 strain D39 that contains an optimized luxABCDE cassette. NanA, SpxB, and, to a lesser extent, CbpA contributed to prolonged nasopharyngeal colonization, whereas CbpA and NanA contributed to the transition to the lower respiratory tract. Once lung infection was established, Pln, SpxB, and LytA contributed to bacterial replication in the lungs and translocation to the bloodstream. In the bloodstream, only Pln and LytA were required for high-titer replication, whereas CbpA was required for invasion of the cerebrospinal fluid. We conclude that transitions between body sites require virulence determinants distinct from those involved in organ-specific replication.


Mitochondrial resolution of a deep branch in the genealogical tree for perching birds.
Animal mitochondrial DNA (mtDNA) is known to contain information about the genealogical relations among closely related species and is shown here to yield information about distant relations as well. Our results also draw attention to the need for caution in using third positions of codons for tree construction. This is evident from comparative studies of the cytochrome b gene in 13 species representing major groups within the order of perching birds (Passeriformes). Sequences of a 924 base-pair segment of this gene were obtained from each of these species via the polymerase chain reaction and a novel set of versatile primers. With a woodpecker sequence as an outgroup, trees that separate songbirds from other perching birds and resolve the ancient branch leading to songbirds were obtained utilizing the conservative first and second positions of codons. Analysis of positions within codons suggests that, for deep branches, the skewed base composition at the fast-changing third positions can result in phylogenetic disinformation, which conflicts with the information retained in the first and second positions. The mitochondrial tree shows broad concordance with that based on hybridization of nuclear DNA; however, parsimony and maximum likelihood methods suggest a close kinship between thrushes and Australian babblers, in agreement with the traditional morphological classification.


Isolation and sequencing of a novel tropomyosin isoform preferentially associated with colon cancer.
Nonmuscle human tropomyosin (hTM) isoforms have distinct functions and may play important roles in various disease processes. In an attempt to identify colon epithelial tropomyosin isoform, a complementary DNA library prepared from a human colon cancer cell line T84 was screened by an oligonucleotide probe complementary to messages of all known hTM isoforms. A novel clone called TC22 was obtained. The amino acid sequence of TC22 isoform is identical to isoform 5 (hTM5) apart from the C terminal domain, amino acids 222-247 coding the exon 9. Northern blot analysis showed that TC22 message is expressed in transformed epithelial cell lines and tumor tissues but not in normal epithelial cells. We developed a monoclonal antibody specific to TC22 isoform (TC22-4). By Western blot and immunoperoxidase assays, we analyzed 105 colonic specimens (fresh frozen and formalin fixed) from 96 patients with colon polyps (hyperplastic) or adenomas with or without dysplasia and cancer. Twenty-one of 22 (95%) of colon cancer specimens showed the presence of TC22, compared with only 1 of the 17 normal colon specimens and none of the 13 hyperplastic polyps (P < 0.0001). As assayed by immunoperoxidase staining, TC22 expression progressively increased in benign adenomatous polyps (35%) and polyps with mild and severe dysplasia (57% and 100%, respectively). We cloned and sequenced a novel hTM isoform, TC22, which is strongly associated with colonic neoplasia and carcinoma. TC22 may provide a useful biomarker for surveillance of colon cancer.


Effect of germination time on antioxidative activity and composition of yellow pea soluble free and polar soluble bound phenolic compounds.
This research aims to study antioxidative activities of polar solvent extractable phenolic compounds from yellow peas with different germination times against oil-in-water emulsion oxidation. After germination (0, 2, 4, and 6 days), soluble free and polar soluble bound phenolic compounds were extracted and their antioxidative activity was evaluated using stripped soybean oil (SSO)-in-water emulsions. Liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS) and size-exclusion chromatography with multiangle-light-scattering and refractive-index detection (SEC-MALS-RI) were employed to analyze the phenolic composition and molar mass, respectively. Antioxidative activities of soluble free phenolic compounds increased in the SSO-in-water emulsion system, while those of polar soluble bound phenolic compounds decreased with germination. On the basis of chemometric analysis, pratensein (2), phloridzin (4), quercetin (9), sayanedine (12), hesperetin (13), glyzaglabrin (14), and pinocembrin (15) were speculated as the pivotal phenolic compounds responsible for the hydrogen donating capacity. Additionally, decreased molecular weight of soluble bound phenolic compounds was accompanied by the reduction of antioxidative activity in SSO-in-water emulsions indicating that the moieties of polar soluble bound phenolic compounds also have an important impact on the antioxidative activity of phenolic compounds.


Fecal microbiota transplantation for the management of autoimmune diseases: Potential mechanisms and challenges.
Autoimmune diseases (AIDs) are a series of immune-mediated lethal diseases featured by over-activated immune cells attacking healthy self-tissues and organs due to the loss of immune tolerance, which always causes severe irreversible systematical organ damage and threatens human health heavily. To date, there are still no definitive cures for the treatment of AIDs due to their pathogenesis has not been clearly understood. Besides, the current clinical treatments of AIDs majorly rely on glucocorticoids and immune suppressors, which can lead to serious side effects. In the past years, there are increasing studies demonstrating that an imbalance of gut microbiota is intimately related to the pathogenesis of various AIDs, shedding light on the development of therapeutics by targeting the gut microbiota for the management of AIDs. Among all the approaches targeting the gut microbiota, fecal microbiota transplantation (FMT) has attracted increasing interest, and it has been proposed as a possible strategy to intervene in the homeostasis of gut microbiota for the treatment of various diseases. However, despite the reported good curative effects and clinical studies conducted on FMT, the detailed mechanisms of FMT for the effective treatment of those diseases have not been figured out. To fully understand the mechanisms of the therapeutic effects of FMT on AIDs and improve the therapeutic efficacy of FMT treatment, a systematic review of this topic is necessary. Hence, in this review paper, the potential mechanisms of FMT for the treatment of various AIDs were summarized, including promotion, shaping, activation, or inhibition of the host immune system via the interactions between the microorganisms and the gut immune system, gut-brain, gut-liver, gut-kidney axis, and so on. Then, applications of FMT for the treatment of various AIDs were detailed presented. Finally, the current challenges and potential solutions for the development of FMT formulations and FMT therapeutics were comprehensively discussed.


Microbiological/clinical characteristics and validation of topical therapy with kanamycin in aerobic vaginitis: a pilot study.
The term 'aerobic vaginitis' defines a 'new' vaginal pathology that is neither classifiable as specific vaginitis nor as bacterial vaginosis. We studied a sample of 30 women with a clinical and microbiological diagnosis of aerobic vaginitis and compared the efficacy and tolerability of kanamycin and meclocycline, two products commercially available in Italy in the form of vaginal pessaries. In chronological order of enrollment, the patients were alternately treated with kanamycin or meclocycline; the dose of administration in both groups was of one pessary per day for 6 days. The evaluation of the therapeutic efficacy was carried out both at the first check-up (7th-8th day) and at a second check-up (13th-16th day). At the first follow-up carried out immediately at the end of therapy, the percentage of normalisation of clinical signs and symptoms was increased independently of the type of treatment in the case of moderate grade aerobic vaginitis, while kanamycin was produced a better effect in the group with severe aerobic vaginitis. Furthermore, at the second follow-up, a direct correlation with recovery of vaginal homeostasis was demonstrated by the normalisation of the vaginal pH and by the presence of lactobacilli, only in kanamycin treated group. In conclusion, our results showed the validity of the treatment with kanamycin intravaginally in this recently recognised disease.


Patterning the heart's left-right axis: from zebrafish to man.
Normal left-right asymmetry is highly conserved among vertebrates. Errors in the proper patterning of this axis are believed to lead to congenital anomalies of the heart and abdominal viscera, often with profound clinical consequences. We review briefly the nature of potential signals and signaling sources that lead to the break in left-right symmetry. The evidence suggests that left-right reversal, or homogenization, of these signals may lead to different consequences, and we explain some malpositions and malalignments of the atria, ventricles, and/or outflow tract that are seen in a variety of congenital cardiac diseases. We speculate that there are units of organ assembly responsive to laterality signals, and these units may be driven independently. One crucial source of signals appears to be the notochord and floorplate. In order to examine the clinical relationship of these midline structures to putative disorders of laterality, we review all patients with disturbances of normal laterality seen at the Massachusetts General Hospital over the past 20 years. We find a significant association between laterality defects and anomalies of the spine and other midline structures.


Fluoxetine ameliorates imbalance of redox homeostasis and inflammation in an acute kidney injury model.
Ischemia-reperfusion (IR) has been reported to be associated with augmented reactive oxygen radicals and cytokines. Currently, we aimed to examine the influence of fluoxetine, which is already used as a preoperative anxiolytic, in the context of IR induced by occlusion of infrarenal abdominal aorta (60 min of ischemia) and its effects on renal oxidative status, inflammation, renal function, and cellular integrity in reperfusion (120 min post-ischemia). Male rats were randomly assigned as control, IR, and pretreated groups. The pretreated group animals received fluoxetine (20 mg/kg, i.p.) once daily for 3 days. Renal tissue oxidative stress, myeloperoxidase activity, proinflammatory cytokines (tumor necrosis factor-α, interleukin-1β, interleukin-6), histology, and function were assessed. As an anti-inflammatory cytokine, interleukin-10 was also assessed. IR led to a significant increase in lipid hydroperoxide, malondialdehyde, and pro-oxidant antioxidant balance and decrease in superoxide dismutase activity and ferric reducing/antioxidant power level (p < 0.05), but fluoxetine was able to restore these parameters. High concentrations of tumor necrosis factor-α, interleukin-1β, interleukin-6, and myeloperoxidase activity caused by IR were significantly decreased in kidney tissue with fluoxetine. In addition, interleukin-10 levels were high in fluoxetine pretreated group. IR resulted in disrupted cellular integrity, infiltration of tissue with leukocytes, and decreased serum creatinine-urea levels (p < 0.05). Fluoxetine significantly restored impaired redox balance and inflammation parameters of rats subjected to IR to baseline values. This beneficial effect of fluoxetine on redox balance might be addressed to an improvement in renal function.


Caspase-3 and caspase-7 but not caspase-6 cleave Gas2 in vitro: implications for microfilament reorganization during apoptosis.
Apoptosis is characterized by proteolysis of specific cellular proteins by a family of cystein proteases known as caspases. Gas2, a component of the microfilament system, is cleaved during apoptosis and the cleaved form specifically regulates microfilaments and cell shape changes. We now demonstrate that Gas2 is a substrate of caspase-3 but not of caspase-6. Proteolytic processing both in vitro and in vivo is dependent on aspartic residue 279. Gas2 cleavage was only partially impaired in apoptotic MCF-7 cells which lack caspase-3, thus indicating that different caspases can process Gas2 in vivo. In vitro Gas2 was processed, albeit with low affinity, by caspase-7 thus suggesting that this caspase could be responsible for the incomplete Gas2 processing observed in UV treated MCF-7 cells. In vivo proteolysis of Gas2 was detected at an early stage of the apoptotic process when the cells are still adherent on the substrate and it was coupled to the specific rearrangement of the microfilament characterizing cell death. Finally we also demonstrated that Gas2 in vitro binds to F-actin, but this interaction was unaffected by the caspase-3 dependent proteolytic processing.


[The SLAM-associated protein (SAP) regulates IFN-gamma expression in leprosy].
The SLAM-associated protein (SAP) regulates IFN-gamma expression in leprosy. Tuberculoid leprosy patients locally produce Th1 cytokines, while lepromatous patients produce Th2 cytokines. Signaling lymphocytic activation molecule (SLAM) and the SLAM-associated protein (SAP) participate in the differentiation process that leads to the production of specific patterns of cytokines by activated T cells. To investigate the SLAM/SAP pathway in M. leprae infection, we determined the expression of SAP, IFN-gamma and SLAM RNA messenger in leprosy patients. We found a direct correlation of SLAM expression with IFN-gamma expression, whereas the expression of SAP was inversely correlated with the expression of both SLAM and IFN-gamma. Therefore, our data indicate that SAP might interfere with Th1 cytokine responses while SLAM expression may contribute to Th1 responses in leprosy. This study further suggests that the SLAM/SAP pathway might be a focal point for therapeutic modulation of T cell cytokine responses in diseases characterized by dysfunctional Th2 responses.


Endovascular treatment with flow diverters of recanalized and multitreated aneurysms initially treated by endovascular approach.
To evaluate the feasibility, safety and efficacy of endovascular treatment with flow diverters in patients with recanalized and multitreated aneurysms in a retrospective, multicenter, single-arm study. The study included 29 patients with 29 recanalized aneurysms who were treated by flow diverters (Silk or Pipeline devices). Pre- and post-procedural complications and morbidity and mortality rates were evaluated and functional outcomes (modified Rankin Score (mRS)) at 1 month (short-term) and 3-4 months (mid-term) were compared with preoperative mRS (before the procedure). Mid-term angiographic follow-up was performed assessing aneurysmal occlusion by the Montreal scale (complete occlusion, neck remnant, aneurysm remnant). Placement of the flow diverters was achieved in all patients. Two misdeployments of the flow diverters necessitated balloon dilation in two patients, which was associated with stent delivery in one patient. Permanent morbidity related to treatment was 6.9% (2/29), transient morbidity was 10.3% (3/29) and there were no deaths resulting from the treatment. One patient died from a myocardial infarct 4 weeks after the procedure. 25/29 patients (86.2%) had a good final functional outcome, 26/29 (89.7%) had an unchanged functional outcome and 2/29 patients (6.9%) had clinical worsening. Angiographic follow-up showed complete occlusion in 17/28 patients (60.7%), neck remnants in 6/28 patients (21.4%) and residual aneurysms in 5/28 (17.9%). Flow diverter placement is feasible and safe in patients with recanalized and multitreated aneurysms. The procedure is associated with a high percentage of good functional outcomes as well as good mid-term anatomical results (82.1%).


Complete jejunoileal necrosis due to torsion of the superior mesenteric artery.
We have reported a case of small-bowel volvulus in which complete jejunoileal necrosis resulted from torsion of the superior mesenteric artery. This case was unusual not only because of the extent of necrosis but also because primary small-bowel volvulus is rare in adults. Despite nearly total small-bowel resection, the patient continues to do well 6 months postoperatively.


Impact of Routine and Long-Term Follow-Up on Weight Loss after Bariatric Surgery.
Weight loss after bariatric surgery varies among patients. Patients who do not complete long-term follow-up are considered to loose less weight than those with regular follow-up visits. To evaluate the influence of patients' follow-up compliance on long-term excess weight loss (%EWL) and total weight loss (%TWL) after bariatric surgery, comparing results between gastric bypass (GB) and sleeve gastrectomy (SG). Patients with up to 5 years of follow-up data after bariatric surgery were included in this retrospective analysis. Patients were divided in 2 groups: those in group 1 who had attended every scheduled postoperative appointment and those in group 2 who had been lost to follow-up before 1 year and were later contacted by telephone. %EWL and %TWL were compared to determine the possible relationship between type of surgery and regularity of the follow-up. A total of 385 patients were included. A significant difference in EWL was observed at 5 years in the SG group (78% for group 1 versus 39% for group 2; p = 0.02) and GB group (75% for group 1 versus 62% for group 2; p = 0.01). No significant differences between surgeries were found when comparing long-term EWL in group 1 patients 77% for SG versus 75% for GB. For group 2 patients, GB achieved greater EWL than SG; p = 0.005. %TWL patients in group 2 showed significant differences in all periods of study (p < 0.05). Bariatric surgery patients who attended all scheduled follow-up appointments experienced significantly greater long-term EWL and TWL than those who did not. GB has apparent increased benefits for weight loss in long-term follow-up when compared with SG for patients who did not attend long-term follow-up. Therefore, continued long-term follow-up of bariatric patients should be encouraged to increase postoperative weight loss results.


Voltage-gated Ca2+ conductances in acutely isolated guinea pig dorsal cochlear nucleus neurons.
Although it is known that voltage-gated Ca2+ conductances (VGCCs) contribute to the responses of dorsal cochlear nucleus (DCN) neurons, little is known about the properties of VGCCs in the DCN. In this study, the whole cell voltage-clamp technique was used to examine the pharmacology and voltage dependence of VGCCs in unidentified DCN neurons acutely isolated from guinea pig brain stem. The majority of cells responded to depolarization with sustained inward currents that were enhanced when Ca2+ was replaced by Ba2+, were blocked partially by Ni2+ (100 microM), and were blocked almost completely by Cd2+ (50 microM). Experiments using nifedipine (10 microM), omegaAga IVA (100 nM) and omegaCTX GVIA (500 nM) demonstrated that a variety of VGCC subtypes contributed to the Ba2+ current in most cells, including the L, N, and P/Q types and antagonist-insensitive R type. Although a large depolarization from rest was required to activate VGCCs in DCN neurons, VGCC activation was rapid at depolarized levels, having time constants <1 ms at 22 degrees C. No fast low-threshold inactivation was observed, and a slow high-threshold inactivation was observed at voltages more positive than -20 mV, indicating that Ba2+ currents were carried by high-voltage activated VGCCs. The VGCC subtypes contributing to the overall Ba2+ current had similar voltage-dependent properties, with the exception of the antagonist-insensitive R-type component, which had a slower activation and a more pronounced inactivation than the other components. These data suggest that a variety of VGCCs is present in DCN neurons, and these conductances generate a rapid Ca2+ influx in response to depolarizing stimuli.


Characterizing the immiscible transport properties of diesel and water in peat soil.
Extensive pipeline and railway corridors crossing Canadian peatlands make them vulnerable to hydrocarbon spills, potentially impairing ecosystem health, so it is important to be able to forecast hydrocarbon fate and transport within and beyond the peatland. The redistribution of hydrocarbon liquids in groundwater systems are controlled by the multiphase flow characteristics of the aquifer material including capillary pressure-saturation-relative permeability (Pc-S-kr) relations. However, these relations have never been characterized for the hydrocarbon-water phases in peat. To address this, the flow and transport of diesel and water in peat soils were examined through a number of one dimensional vertical immiscible displacement tests, in which diesel was percolated into peat pore space displacing peat water, leading to a two-phase flow regime. Inverse modelling simulations using both Brooks and Corey's and power law relative permeability models, matched the data of the immiscible displacement tests well. Irreducible water saturation (Swirr) and the curvature of water relative permeability relation increased with peat bulk density. The residual diesel saturation (SNr) ranged between 0.3% and 17% and increased with bulk density of peat. In a given peat, SNr was a function of saturation history and increased with increasing maximum diesel saturation. The receding contact angles of water in water-air systems and diesel in diesel-air systems, respectively, were 51.7° and 61.2°, showing that the wetting tendency of peat in the air imbibition condition is toward the draining liquid. These experiments advance our knowledge on the behavior of hydrocarbons in peat, and can improve numerical modelling of hydrocarbon transport after a spill.


Simulation-based education and performance assessments for pediatric surgeons.
Education in the knowledge, skills, and attitudes necessary for a surgeon to perform at an expert level in the operating room, and beyond, must address all potential cognitive and technical performance gaps, professionalism and personal behaviors, and effective team communication. Educational strategies should also seek to replicate the stressors and distractions that might occur during a high-risk operation or critical care event. Finally, education cannot remain fixed in an apprenticeship model of "See one, do one, teach one," whereby patients are exposed to the risk of harm inherent to any learning curve. The majority of these educational goals can be achieved with the addition of simulation-based education (SBE) as a valuable adjunct to traditional training methods. This article will review relevant principles of SBE, explore currently available simulation-based educational tools for pediatric surgeons, and finally make projections for the future of SBE and performance assessments for pediatric surgeons.