Miracle-dz/UniFAR-FTUs huggingface dataset:
The Pseudomonas aeruginosa Exotoxin A Regulatory Gene, ptxS: Evidence for Negative Autoregulation

ABSTRACT We have previously described a Pseudomonas aeruginosagene, ptxR, which enhances exotoxin A production at the transcriptional level. We have also described another gene,ptxS, which is transcribed divergently fromptxR and interferes with the enhancement of exotoxin A synthesis by ptxR. However, the mechanisms through whichptxR and/or ptxS are regulated is not known. In this study, we attempted (by using the DNA gel shift assay) to determine if P. aeruginosa contains a potential regulatory protein that binds specifically to the ptxR orptxS upstream region. In the initial analysis, different-sized gel shift bands were detected when a probe containing the ptxR-ptxS intergenic region was incubated with the lysate of P. aeruginosa PAO1. The strongest binding activity was detected with a smaller fragment that represents theptxS upstream region. Additional deletion analysis localized the binding to a 52-bp fragment immediately upstream ofptxS. The gel shift band was not detected when the 52-bp fragment was incubated with the lysate of the ptxS isogenic mutant PAO1::ptxS. However, the binding band was regenerated when a plasmid carrying ptxS intact was introduced into PAO1::ptxS. In addition, the gel shift band was detected when the 52-bp fragment was incubated with a lysate of Escherichia coli in which ptxS was overexpressed from the T7 promoter. The effect of PtxS onptxS expression was examined by using aptxS-lacZ fusion plasmid. The level of β-galactosidase activity produced by PAO1::ptxS carrying the fusion plasmid was four- to fivefold higher than that produced by PAO1 carrying the same plasmid. Using DNase I footprinting analysis, the binding region was specified to a 20-bp fragment. Within the fragment, a 14-bp palindromic sequence exists that may function as a PtxS binding site. These results suggest that PtxS autoregulates its synthesis by binding to a specific sequence within the ptxSupstream region.
Iron fortified follow on formula from 9 to 18 months improves iron status but not development or growth: a randomised trial

AIMS Iron deficiency anaemia is associated, in observational studies, with developmental disadvantage. This study tested the hypothesis that feeding iron supplemented formula from 9 to 18 months of age would improve developmental performance. SUBJECTS AND METHODS 493 healthy children aged 9 months being fed pasteurised cows’ milk were recruited from three UK centres. They were randomised to: cows’ milk as before, formula containing 0.9 mg/litre iron, or formula containing 1.2 mg/litre iron, until 18 months of age. Bayley mental and psychomotor developmental indices were measured at 18 months, as were growth and haematological indices. RESULTS Children fed iron fortified formula had higher plasma ferritin concentrations, but there were no significant intergroup differences in development or growth. CONCLUSIONS There are no developmental or growth advantages in children given iron supplemented formula, but a benefit for a minority who were anaemic, or the possibility that a benefit may emerge at a later age, cannot be excluded.
Preclinical evaluation and validation of [18F]HX4, a promising hypoxia marker for PET imaging

Hypoxia has been shown to be an important microenvironmental parameter influencing tumor progression and treatment efficacy. Patient guidance for hypoxia-targeted therapy requires evaluation of tumor oxygenation, preferably in a noninvasive manner. The aim of this study was to evaluate and validate the uptake of [18F]HX4, a novel developed hypoxia marker for PET imaging. A heterogeneous accumulation of [18F]HX4 was found within rat rhabdomyosarcoma tumors that was significantly (P < 0.0001) higher compared with the surrounding tissues, with temporal increasing tumor-to-blood ratios reaching a plateau of 7.638 ± 0.926 and optimal imaging properties 4 h after injection. [18F]HX4 retention in normal tissues was found to be short-lived, homogeneous and characterized by a fast progressive temporal clearance. Heterogeneity in [18F]HX4 tumor uptake was analyzed based on 16 regions within the tumor according to the different orthogonal planes at the largest diameter. Validation of heterogeneous [18F]HX4 tumor uptake was shown by a strong and significant relationship (r = 0.722; P < 0.0001) with the hypoxic fraction as calculated by the percentage pimonidazole-positive pixels. Furthermore, a causal relationship with tumor oxygenation was established, because combination treatment of nicotinamide and carbogen resulted in a 40% reduction (P < 0.001) in [18F]HX4 tumor accumulation whereas treatment with 7% oxygen breathing resulted in a 30% increased uptake (P < 0.05). [18F]HX4 is therefore a promising candidate for noninvasive detection and evaluation of tumor hypoxia at a macroscopic level.
Integrin Expression and Function on Human Osteoblast‐like Cells

The integrin family of cell adhesion molecules are a series of cell surface glycoproteins that recognize a range of cell surface and extracellular matrix (ECM)‐associated ligands. To date, the precise role of individual integrin molecules in bone cell–ECM interactions remains unclear. Cell binding assays were performed to examine the ability of normal human bone cells (NHBCs) to adhere to different ECM proteins in vitro. NHBCs displayed preferential adhesion to fibronectin over collagen types I, IV, and vitronectin and showed low affinity binding to laminin and collagen type V. No binding was observed to collagen type III. The integrin heterodimers α1β1, α2β1, α3β1, α5β1, αvβ3, and αvβ5 were found to be constitutively expressed on the cell surface of NHBCs by flow cytometric analysis. The integrins α4β1 and α6β1 were not expressed by NHBCs. Subsequent binding studies showed that NHBC adhesion to collagen and laminin was mediated by multiple integrins where cell attachment was almost completely inhibited in the presence of a combination of function‐blocking monoclonal antibodies (Mabs) to α1β1, α2β1, α3β1, and β1. In contrast, the adhesion of NHBCs to fibronectin was only partially inhibited (50%) in the presence of blocking Mabs to α3β1, α5β1, and β1. The attachment of NHBCs to collagen, laminin, fibronectin, and vitronectin was also found to be unaffected in the presence of a function‐blocking Mab to αvβ3. The results of this study indicate that β1 integrins appear to be the predominant adhesion receptor subfamily utilized by human osteoblast‐like cells to adhere to collagen and laminin and in part to fibronectin.
Postischemic blood flow response in hypercholesterolemic patients.

We undertook this cross-sectional study to compare the mechanical behavior and postischemic response of the radial artery of 15 newly diagnosed hypercholesterolemic patients with those of 15 age- and sex-matched normocholesterolemic control subjects and 21 hypercholesterolemic patients treated for 2 years with an 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor (simvastatin, 10 to 20 mg/d). At the time of the study total cholesterol levels were at 7.9 +/- 0.2, 4.9 +/- 0.2, and 6.0 +/- 0.3 mmol/L in the three groups, respectively (mean +/- SEM, P < .001). High-resolution, noninvasive echotracking for assessment of internal arterial diameter was combined with measurements of blood flow velocity by Doppler and blood pressure by photoplethysmography. Radial cross-sectional compliance and distensibility were similar in all groups. Forearm blood flow and flow-mediated dilation were measured after a 5-minute upper arm occlusion. Flow was calculated from the vessel diameter and blood flow velocity recorded simultaneously at the same site. Flow-mediated dilation after ischemia was not significantly different among the three groups. However, forearm blood flow increase was markedly blunted (P < .01) in untreated hypercholesterolemic patients (211%) compared with the normocholesterolemic control subjects (411%) and treated patients (365%). These findings suggest that the distensibility of the radial artery, a muscular conduit vessel usually devoid of atherosclerotic lesions, and its flow-mediated dilation are preserved in hypercholesterolemic patients. In contrast, forearm resistance vessels exhibit a markedly reduced postischemic blood flow response that may be restored by prolonged lipid-lowering intervention.
Major Urinary Protein as a Negative Tumor Marker in Mouse Hepatocarcinogenesis

A cDNA clone isolated by differential screening of a mouse liver cDNA library using RNA probes from normal adult liver and from hepatocellular tumors showed, on northern blot analysis, a single transcript of 1 kb present at high levels in normal adult mouse liver but at 15‐ to 100‐fold reduced levels in spontaneous and p‐dimethylaminoazobenzene‐ or urethan‐induced liver tumors. The transcript, which was absent in two‐wk‐old mouse liver, is coded by a developmentally regulated gene. Nucleotide sequence analysis demonstrated that the isolated clone encoded a major urinary protein (MUP). Immunocytochemistry analysis, using a goat antibody to MUP on histologic liver sections containing diethylnitrosamine‐induced liver nodules, showed that the decrease in MUP levels also occurred in microscopic nodules. Therefore, MUP decrease represents an early event in the development of mouse liver tumors, and it may be used as a tumor marker in mouse hepatocarcinogenesis.
Individual acid aspartic proteinases (Saps) 1-6 of Candida albicans are not essential for invasion and colonization of the gastrointestinal tract in mice.

In order to investigate whether there is a role for individual secreted aspartic proteinases (Saps) of Candida albicans in gastrointestinal infection of mice we compared the differential expression of SAP1-6 genes and production of Sap1-6 proteins with invasion and persistence of SAP knockout strains in the gastrointestinal tract. Using an in vivo expression technology (IVET) we found a high percentage of expression of SAP4-6 genes which increased steadily in the course of infection. Expression of SAP1-3 genes was detected occasionally and in lower percentages than that of SAP4-6 genes. With reverse transcriptase-polymerase chain reaction (RT-PCR), mRNA for SAP 4 and SAP6 were detected in the stomach of all mice, whereas SAP2, SAP3 and SAP5 mRNA were detected not in all animals and SAP1 mRNA was not detectable. Also with immunoelectron microscopy we demonstrated production of Saps1-3 as well as Saps4-6 with antibodies cross-reacting with either Saps1-3 or Saps4-6. In contrast to the fact that gene expression and production of Saps were readily detectable, we were unable to demonstrate differences in the ability to invade the stomach, to disseminate to the brain as well as in the duration of faecal shedding and the number of fungi persisting in the faeces of mice infected with SAP knockout strains in comparison to control strains. We conclude that although Saps were produced, individual Saps were not indispensable factors for virulence during gastrointestinal infection of mice.
Mobile-Bearing Total Knee Arthroplasty: Do the Polyethylene Bearings Rotate?

In vivo kinematics were determined during a weightbearing deep knee bend in 39 patients implanted with four different designs of mobile-bearing total knee arthroplasty to assess polyethylene bearing mobility patterns and magnitudes. The femoral and tibial components and mobile polyethylene insert (implanted with four tantalum beads) were overlaid onto the fluoroscopic images using a three-dimensional model-fitting technique to determine bearing mobility. Three of the four designs were tested at a single time interval while one was evaluated at two postoperative intervals, (12 months apart) to assess changes in bearing mobility with time. All patients had polyethylene bearing rotation relative to the tibial tray and minimal rotation relative to the femoral component. The average maximum amounts of bearing rotation ranged from 8.4° to 10.3° (range, 3.0°-20.1°). In patients evaluated at two time intervals, the average maximum bearing rotation increased from 8.5° (range, 3.4°-15.5°) at 3 months to 9.8° (range, 48°-14.1°) 15 months postoperatively. The presented data demonstrates polyethylene bearing mobility occurs following mobile-bearing total knee arthroplasty and mobility is maintained during the time interval tested. The presence of bearing mobility should result in lower contact stresses reducing the potential for polyethylene wear.
Maintenance efficacy designs in psychiatry: Randomized discontinuation trials – enriched but not better

Introduction Although classical randomized clinical trials (RCTs) are the gold standard for proof of drug efficacy, randomized discontinuation trials (RDTs), sometimes called “enriched” trials, are used increasingly, especially in psychiatric maintenance studies. Methods A narrative review of two decades of experience with RDTs. Results RDTs in psychiatric maintenance trials tend to use a dependent variable as a predictor: treatment response. Treatment responders are assessed for treatment response. This tautology in the logic of RDTs renders them invalid, since the predictor and the outcome are the same variable. Although RDTs can be designed to avoid this tautologous state of affairs, like using independent predictors of outcomes, such is not the case with psychiatric maintenance studies Further, purported benefits of RDTs regarding feasibility were found to be questionable. Specifically, RDTs do not enhance statistical power in many settings, and, because of high dropout rates, produce results of questionable validity. Any claimed benefits come with notably reduced generalizability. Conclusions RDTs appear to be scientifically invalid as used in psychiatric maintenance designs. Their purported feasibility benefits are not seen in actual trials for psychotropic drugs. There is warrant for changes in federal policy regarding marketing indications for maintenance efficacy using the RDT design.
Effects of nitrogen and helium at pressure on drug-induced stereotypy and locomotion in mice.

Stereotyped behavior was induced in mice by apomorphine (5-50 mg X kg-1 s.c.) and d-amphetamine (3-5 mg X kg-1 i.p.), and increased locomotor activity was induced by d-amphetamine (8 and 10 mg X kg-1 i.p.) and by morphine sulfate (15 and 30 mg X kg-1 i.p.). Experiments were conducted at 1, 4, and 7 ATA. All mice were myringotomized under ether anesthesia 3 d before behavioral studies to minimize disturbances due to pressure differences across the tympanic membrane during compression. Compressed air significantly increased locomotor activity induced by d-amphetamine and morphine sulfate whereas He/O2 had no effect, suggesting that the change was due to the narcotic effect of N2. Drug-induced stereotyped behavior was affected variably (usually depressed) by compressed air and not by He/O2. Since both stereotypy and locomotor activity induced by these drugs involve dopamine receptor systems, the results suggest that compressed air does not influence all such membrane receptors in like manner. Evidence for receptor plasticity is discussed.
Simultaneous non-invasive measurement of liquid gastric emptying and small bowel transit by combined 13C-acetate and H2-lactulose breath test

The 13C-acetate breath test (13C-ABT) allows noninasive measurement of liquid gastric emptying time (LGET), the H2-lactulose breath (H2-LBT) measures orocecal transit time (OCTT). Because of different test principles, both tests can generally be combined. This would not only spare time and resources but may also deliver additional information on the integrated regulation of gastrointestinal motor functions. Our aim was to define an adequate test protocol and to generate normal values for the combined 13C-ABT/H2-LBT. Twenty two healthy volunteers participated (HV, 12 females, age: 26.5 ± 4.7 years, BMI: 22 ± 2.1 kg m−2). Fasting subjects received 200 ml of an aequous solution containing 150 mg 13C-acetate and 10 g lactulose. Breath samples for H2 measurements were collected before ingestion of the test solution and afterwards at 5 min intervals for 180 min, for 13C measurements sampling intervals were prolonged to 15 min after the first hour. LGET was defined as time of maximal 13C-exhalation, OCTT as increase in H2 of ⩾ 20 ppm above basal. Small bowel transit time (SBTT) was calculated as the difference between OCTT and LGET. Mean [95% CI] values for LGET, OCTT and SBTT were 26 [23–30] min, 85 [68–102] min and 60 [44–76] min, respectively. Normal values for LGET and OCTT corresponded with published data. SBTT was not correlated with LGET (R2 = 0.019; p = 0.546). By contrast, there was a very tight and highly significant correlation between SBTT and OCTT (R2 = 0.933; p < 0.001). Multivariate linear regression analyses confirmed that age, sex and BMI had no significant effects on OCTT or SBTT while LGET was weakly correlated with BMI (R2 = 0.240; p = 0.021). In conclusion, the combined 13C-ABT/H2-LBT allows simultaneous measurement of LGET and OCTT as well as calculation of SBTT.
Temperament and character traits in major depressive disorder: influence of mood state and recurrence of episodes

Background: The objective of this study was to compare personality traits between major depressive disorder (MDD) patients and healthy comparison subjects (HC) and examine if personality traits in patients are associated with specific clinical characteristics of the disorder. Methods: Sixty MDD patients (45 depressed, 15 remitted) were compared to 60 HC using the Temperament and Character Inventory. Analysis of covariance, with age and gender as covariates, was used to compare the mean Temperament and Character Inventory scores among the subject groups. Results: Depressed MDD patients scored significantly higher than HC on novelty seeking, harm avoidance, and self‐transcendence and lower on reward dependence, self‐directedness, and cooperativeness. Remitted MDD patients scored significantly lower than HC only on self‐directedness. Comorbidity with anxiety disorder had a main effect only on harm avoidance. Harm avoidance was positively correlated with depression intensity and with number of episodes. Self‐directedness had an inverse correlation with depression intensity. Conclusions: MDD patients present a different personality profile from HC, and these differences are influenced by mood state and comorbid anxiety disorders. When considering patients who have been in remission for some time, the differences pertain to few personality dimensions. Cumulated number of depressive episodes may result in increased harm avoidance. Depression and Anxiety, 2009. © 2009 Wiley‐Liss, Inc.
Quantification of Physical Activity and Sedentary Time in Adults with Cerebral Palsy.

PURPOSE The purpose of this study was to determine objective and subjective quantification of habitual physical activity (HPA) and sedentary time in ambulatory and nonambulatory adults with cerebral palsy (CP). METHODS We recruited a clinical sample of adults with CP (N = 42; 21 women; mean (SD) age, 33.5 (12.3) yr; Gross Motor Function Classification System (GMFCS) distribution: level I (n = 5), level II (n = 9), level III (n = 10), level IV (n = 11), and level V (n = 7). Objective measures of HPA and sedentary time were obtained by using ActiGraph GT3X accelerometers at both hip and wrist sites. Three previously established cut-point values distinguishing light physical activity (LPA) and moderate-to-vigorous physical activity (MVPA) were evaluated across GMFCS levels. The concurrent validity of the self-report Physical Activity Recall Assessment for People with Spinal Cord Injury (PARA-SCI) was assessed for LPA and MVPA intensities in GMFCS levels II-V. RESULTS Participants showed little reluctance to wearing accelerometers; one participant reported discomfort. Nonambulatory adults (GMFCS levels IV-V) differed from ambulatory adults (GMFCS levels I-III) for recorded activity counts (hip and wrist sites), minutes of MVPA with each cut-point value, and breaks from sedentary time (all P < 0.05). For the same measures, adults in GMFCS level III also differed from GMFCS level I (all P < 0.05). The PARA-SCI correlated significantly with accelerometer-derived minutes of MVPA per day (r = 0.396; P = 0.014) and per hour of monitoring time (r = 0.356; P = 0.027). CONCLUSIONS Our findings support the use of accelerometers to objectively measure HPA and sedentary behavior in adults with CP across the severity spectrum, regardless of cut-point implementation. The PARA-SCI is a valid tool to capture subjectively reported patterns of MVPA in adults with CP who are GMFCS levels II-V.
Decision criteria for resolving isotype switching conflicts by B cells

Isotype switching by B cells is highly regulated by a group of cytokines including IL‐4, IFN‐γ and TGF‐β. A B cell can only express one isotype at a time; however, during an immune response it may be exposed to combinations of stimuli that provide it with conflicting switching instructions. To determine how such cytokine‐induced isotype switch conflicts would be resolved, the responses of B cells exposed to multiple cytokines were investigated. To eliminate complications arising from simultaneous effects of switching cytokines on proliferation, division number was used as a reference framework to monitor switching rate. The results show a clear hierarchy in which IFN‐γ is dominant over IL‐4, and both IL‐4 and IFN‐γ are dominant over TGF‐β. These studies reveal how B cells possess a set of logical decision criteria for dealing with pathogens that invoke a range of different stimuli.
Performance evaluation of the Sysmex® XP‐300 in an oncology setting: evaluation and comparison of hematological parameters with the Sysmex® XN‐3000

The Sysmex XP‐300® (XP‐300) is a new, fully automated hematology analyzer, designed to generate complete blood counts (CBC) with 3‐part differential. In our study, the XP‐300 was evaluated as a point‐of‐care (POC) analyzer in an oncology setting. In which blood samples from patients with different pathologies and treatments, affecting hematopoiesis, were analyzed.