Miracle-dz/UniFAR-FTUs huggingface dataset:
Sugar binding induces an outward facing conformation of LacY

According to x-ray structure, the lactose permease (LacY) is a monomer organized into N- and C-terminal six-helix bundles that form a deep internal cavity open on the cytoplasmic side with a single sugar-binding site at the apex. The periplasmic side of the molecule is closed. During sugar/H+ symport, a cavity facing the periplasmic side is thought to open with closure of the inward-facing cytoplasmic cavity so that the sugar-binding site is alternately accessible to either face of the membrane. Double electron–electron resonance (DEER) is used here to measure interhelical distance changes induced by sugar binding to LacY. Nitroxide-labeled paired-Cys replacements were constructed at the ends of transmembrane helices on the cytoplasmic or periplasmic sides of wild-type LacY and in the conformationally restricted mutant Cys-154→Gly. Distances were then determined in the presence of galactosidic or nongalactosidic sugars. Strikingly, specific binding causes conformational rearrangement on both sides of the molecule. On the cytoplasmic side, each of six nitroxide-labeled pairs exhibits decreased interspin distances ranging from 4 to 21 Å. Conversely, on the periplasmic side, each of three spin-labeled pairs shows increased distances ranging from 4 to 14 Å. Thus, the inward-facing cytoplasmic cavity closes, and a cleft opens on the tightly packed periplasmic side. In the Cys-154→Gly mutant, sugar-induced closing is observed on the cytoplasmic face, but little or no change occurs on periplasmic side. The DEER measurements in conjunction with molecular modeling based on the x-ray structure provide strong support for the alternative access model and reveal a structure for the outward-facing conformer of LacY.
SPECIFIC NON-COPLANAR PCB-MEDIATED MODULATION OF BOTTLENOSE DOLPHIN AND BELUGA WHALE PHAGOCYTOSIS UPON IN VITRO EXPOSURE

Contaminant-induced immunosuppression by organochlorines (OC), particularly polychlorinated biphenyls (PCBs), has been suspected as a cofactor in the deaths of thousands of marine mammals. One important innate defense mechanism is phagocytosis, the ability of cells to ingest extracellular macromolecules. The present study was aimed at characterizing the immunomodulatory potential of representative OCs on phagocytosis in bottlenose dolphins and beluga whales. The ability of peripheral blood leukocytes to engulf fluorescent microspheres was evaluated using flow cytometry. The immunomodulatory effects of three non-coplanar PCB congeners, 138, 153, and 180, one coplanar PCB, 169, and 2,3,7,8-TCDD and all possible mixtures (26) were tested upon in vitro exposure. In both species, all mixtures containing at least two non-coplanar PCBs significantly reduced both neutrophil and monocyte phagocytosis, with effects more marked in dolphins than in belugas. Coplanar OCs, on their own or when added to non-coplanar congeners, did not further modulate phagocytosis, suggesting an Ah receptor-independent mechanism. Concentration-response experiments with individual congeners further demonstrated a non-coplanar PCB-induced suppression of phagocytosis, while coplanar congeners produced no consistent effects. Our results suggest simple additive interactions of chemicals in a mixture. However, calculation of toxic equivalency (TEQs) failed to predict the experimentally induced immunomodulatory effects of OCs on dolphin and beluga phagocytosis, confirming the Ah receptor-independent nature of the effects on phagocytosis. Overall, our results suggest that non-AhR mechanisms may explain one facet of immunotoxicity (phagocytosis), something that is not captured using the TEQ approach. This is the first report demonstrating the immunomodulatory effects of OCs on dolphin and beluga phagocytosis, and the first overall demonstration of immunomodulatory effects on phagocytosis mediated specifically by non-coplanar PCBs.
The Effect of the Nonspecific Nitric Oxide Synthase Inhibitor NG-Nitro-l-Arginine Methyl Ester on the Choroidal Compensatory Response to Myopic Defocus in Chickens

Purpose. Chick eyes show rapid compensation to retinal defocus. One component of this mechanism involves changes in the thickness of the choroid: when the retina is exposed to myopic defocus, the choroid thickens, pushing the retina forward; conversely, when the eye is exposed to hyperopic defocus, the choroid thins. The underlying mechanism(s) for these changes are unknown. We tested the hypothesis that nitric oxide might play a role. Methods. We examined the effect of the nonspecific nitric oxide synthase inhibitor NG-nitro-l-arginine methyl ester (l-NAME) on the compensatory choroidal thickening in response to myopic defocus using two visual paradigms: first, in previously form-deprived “recovering” eyes and, second, in eyes wearing +15 D spectacle lenses. l-NAME was injected intravitreally after removal of the diffuser or immediately before putting on the lenses. In addition, we looked at the effect of l-NAME on experimentally thickened choroids (induced by 1 week of recovery from deprivation myopia or 1 week of +15 D lens wear) and on choroids of normal eyes. Eyes were measured using A-scan ultrasonography before the injections and at subsequent intervals for several days. As a control for the injection procedure, eyes with the same visual conditions were injected with saline. Fellow eyes were untreated and uninjected. Results. l-NAME inhibited choroidal thickening in both previously form-deprived eyes (2 vs. 117 &mgr;m; p < 0.001) and eyes wearing +15 D lenses (3 vs. 137 &mgr;m; p < 0.02). The effect was rapid, transient, and dose dependent (ED50, 0.26 &mgr;moles). l-NAME produced thinning in experimentally thickened choroids (recovering: −116 &mgr;m; lenses: −219 &mgr;m) and in normal choroids (−47 &mgr;m) within 7 hours. Conclusions. Nitric oxide may play a role in modulating choroidal thickness. The mechanism is as yet unknown.
Analytical 3-D design of a Transverse Flux Machine with high power factor

Low-speed high-torque applications, e.g. wind energy generation, favor high number of pole solutions. for traditional radial or axial flux machines this leads to an increase in leakage flux while the power output does not increase. For Transverse Flux Machines (TFM) the increase in power output is proportional to the number of poles. However, large flux leakage is also present in TFMs, reducing their power factor and commercial application. Fast and accurate 3-D models are required to model this flux leakage. Finite Element Models (FEM) comprising a large number of elements are commonly used to calculate the 3-D flux density distribution. This approach is very time consuming with hours for a single solution. A fast and more accurate parameterized model is essential to minimize the machine volume while maintaining the required power factor. In this paper, a design approach is presented to obtain a TFM of a minimum volume with a predefined power factor. An analytical 3-D magnetic charge model is used to model a single magnetic period of the TFM. With the required power factor and the obtained flux due to the magnets the coil dimensions are calculated. The performance of the single magnetic period of the TFM is used to determine the full machine dimensions.
The extracellular cytolysin of Vibrio vulnificus: inactivation and relationship to virulence in mice

The ca. 51-kDa extracellular cytolysin of Vibrio vulnificus has been proposed as a virulence factor. We inactivated the structural gene for cytolysin in fully virulent, clinical V. vulnificus strains by both transposon mutagenesis and marker exchange techniques. Inactivation of the cytolysin did not affect virulence in our mouse models. The 50% lethal dose of cytolysin-negative strains was comparable to that of the cytolysin-positive parent strains after intraperitoneal inoculation with and without iron loading. Similar results were obtained after intradermal injection: cytolysin-positive and -negative strains had the same 50% lethal dose and caused comparable tissue damage. While we cannot say that the cytolysin has no effect on the pathogenesis of V. vulnificus infections, its role appears to be of much less importance than are other factors, such as encapsulation.
Quantitative culture of endocervical Chlamydia trachomatis

We examined the number of Chlamydia trachomatis inclusions produced in the initial passage of cell cultures of endocervical specimens from 1,231 women with positive chlamydial cultures who attended a sexually transmitted diseases clinic. Youth, white race, oral contraceptive use, and concurrent infection by Neisseria gonorrhoeae were associated with high chlamydial inclusion counts. Youth, white race, and oral contraceptive use were independent determinants of a high chlamydial inclusion count in women without concurrent gonorrhea but not in women with gonorrhea. Results of our study suggest that the degree of chlamydial excretion from the infected cervix may be influenced by characteristics of the patient being tested and may affect the ability to detect C. trachomatis in different patient groups.
Adoption of the hybrid CTO approach by a single non-CTO operator: procedural and clinical outcomes.

BACKGROUND The feasibility of adopting the "hybrid" approach by a single operator without prior experience in percutaneous coronary intervention (PCI) of chronic total occlusions (CTOs) has not been described. METHODS Consecutive patients who underwent CTO-PCI by a single operator using the "hybrid" approach between 2012 and 2013 formed the analytic cohort. No patient was declined on the basis of angiographic findings. Clinical and angiographic characteristics together with procedural and hospital outcomes are described. RESULTS During the study period, a total of 48 consecutive patients underwent PCI of 50 CTOs. Mean age was 63.4 ± 9.4 years and most patients (83%) were men. The right coronary artery (RCA) was the most commonly treated CTO vessel (54%) and mean J-CTO score was 2.3 ± 1.1. A primary retrograde approach was chosen for 33% of lesions and 40% required use of an epicardial collateral vessel. The primary strategy was effective in 65% of successful cases, 35% required one change in strategy, and 15% requiring two strategy changes. Procedural success rate was 92%. The median number of stents used was 3 (interquartile range [IQR], 2-4] and the total stent length was 73 mm [IQR, 38-96 mm). Mean contrast volume was 356.4 ± 148.3 mL and the mean air kerma radiation exposure was 3.5 ± 2.0 Gy. No patient experienced a major periprocedural complication. CONCLUSION The "hybrid" approach to CTO-PCI can be successfully adopted by a single operator with excellent early procedural success and low complication rates, despite a lack of prior CTO-PCI experience.
Day-to-day variability of maximum walking distance in MS patients can mislead to relevant changes in the Expanded Disability Status Scale (EDSS): average walking speed is a more constant parameter

In this preliminary study we measured maximum walking distance and walking time on four consecutive days in 29 patients with clinically stable multiple sclerosis (MS). Patients were included in the study if they could achieve a maximum unaided walking distance of 100 up to 500 m. Our results showed a certain day-to-day variability of maximum walking distance, in some cases meaning changes up to 1.5 points in the expanded disability status scale (EDSS), which could be misinterpreted as a progression of the disease. Simultaneous measurements of maximum walking time showed a similar variability, unlike the mean walking speed which turned out to be more stable. Our results therefore suggest that scoring of MS patients should not be based on one single measurement of the maximum walking distance. The more reliable parameter appears to be the mean walking speed.
Domoic Acid Neurotoxicity in Cultured Cerebellar Granule Neurons Is Mediated Predominantly by NMDA Receptors that Are Activated as a Consequence of Excitatory Amino Acid Release

Abstract: The participation of NMDA and non‐NMDA receptors in domoic acid‐induced neurotoxicity was investigated in cultured rat cerebellar granule cells (CGCs). Neurons were exposed to 300 µMl‐glutamate or 10 µM domoate for 2 h in physiologic buffer at 22°C followed by a 22‐h incubation in 37°C conditioned growth media. Excitotoxic injury was monitored as a function of time by measurement of lactate dehydrogenase (LDH) activity in both the exposure buffer and the conditioned media. Glutamate and domoate evoked, respectively, 50 and 65% of the total 24‐h increment in LDH efflux after 2 h. Hyperosmolar conditions prevented this early response but did not significantly alter the extent of neuronal injury observed at 24 h. The competitive NMDA receptor antagonist d(−)‐2‐amino‐5‐phosphonopentanoic acid and the non‐NMDA receptor antagonist 2,3‐dihydroxy‐6‐nitro‐7‐sulfamoylbenzo(f)quinoxaline (NBQX) reduced glutamate‐induced LDH efflux totals by 73 and 27%, respectively, whereas, together, these glutamate receptor antagonists completely prevented neuronal injury. Domoate toxicity was reduced 65–77% when CGCs were treated with competitive and noncompetitive NMDA receptor antagonists. Unlike the effect on glutamate toxicity, NBQX completely prevented domoate‐mediated injury. HPLC analysis of the exposure buffer revealed that domoate stimulates the release of excitatory amino acids (EAAs) and adenosine from neurons. Domoate‐stimulated EAA release occurred almost exclusively through mechanisms related to cell swelling and reversal of the glutamate transporter. Thus, whereas glutamate‐induced injury is mediated primarily through NMDA receptors, the full extent of neurodegeneration is produced by the coactivation of both NMDA and non‐NMDA receptors. Domoate‐induced neuronal injury is also mediated primarily through NMDA receptors, which are activated secondarily as a consequence of α‐amino‐3‐hydroxy‐5‐methylisoxazole‐4‐propionate (AMPA)/kainate receptor‐mediated stimulation of EAA efflux.
Rapid enumeration of Escherichia coli in oysters by a quantitative PCR‐ELISA

Direct enumeration of Escherichia coli from oysters was achieved using a polymerase chain reaction (PCR) amplification of the lamB gene coupled with an enzyme‐linked immunosorbent assay (ELISA). Amplified PCR products generated using a digoxigenin‐labelled primer were heat denatured before being quantified by an ELISA. A biotinylated probe immobilized onto streptavidin‐coated microplates was used to capture the digoxigenin‐labelled fragments that were detected with a peroxidase antidigoxigenin conjugate. Subsequent enzymic conversion of substrate gave distinct absorbance differences when assaying oyster samples containing E. coli in the range 10–105 cfu g−1.
Risk behaviours in HIV-positive men who have sex with men participating in an intervention in a primary care setting

Men who have sex with men receiving HIV care reported their sexual behaviours and their intentions, classified according to the Transtheoretical Model of Change, to modify the following behaviours: (1) condom use by partner type and activity type; (2) reduction of partner number; and (3) disclosure of HIV serostatus to partners. Most participants were white (68.8%) or black (29.5%) and were more likely to report unprotected sex with HIV-positive than with serodiscordant partners for most activities. Whites reported more partners than black patients (mean 4.1 versus 2; P < 0.0001) and black participants reported fewer HIV-negative (P = 0.0084) and -unknown status partners (P = 0.00095) than whites. Cocaine/crack use was associated with more sexual partners (P = 0.001) and more frequent unprotected sex with HIV-negative or -unknown status partners (P = 0.036). Readiness to change risk behaviour varied by partner status and type of sexual activity. Understanding patients' risks and their readiness to change behaviours may help providers to promote sexual health.
Time Contracts and Temporal Precision Declines When the Mind Wanders

Our perception of time varies considerably from moment to moment, but how this variability relates to endogenous fluctuations in attentional states has been neglected. Here, we tested the hypothesis that perceptual decoupling during spontaneous mind wandering episodes distorts interval timing. In two studies with different visual subsecond interval timing paradigms, participants judged their attentional state on a trial-by-trial basis. Mind wandering states were characterized by underestimation of temporal intervals and a decline in temporal discrimination. Further analyses suggested that temporal contraction during mind wandering, but not the decline in temporal discrimination, could be attributed in part to attentional lapses. By contrast, we did not find any robust evidence that metacognition pertaining to interval timing was altered during mind wandering states. These results highlight the role of transient fluctuations in attentional states in intraindividual variability in time perception and have implications for the perceptual consequences, behavioral markers, and costs and benefits, of mind wandering.
Sirolimus in association with mycophenolate mofetil induction for the prevention of acute graft rejection in renal allograft recipients.

INTRODUCTION A previous trial in renal transplantation comparing sirolimus (rapamycin) to cyclosporine (CsA) in a triple-drug therapy regimen with azathioprine and corticosteroids found that the incidence of acute rejection was similar (approximately 40%) with a trend for better renal function with sirolimus. METHODS In 14 European centers, first cadaveric renal allograft recipients were randomized to receive sirolimus (n = 40) or CsA (n = 38) in an open-label design. All patients received corticosteroids and mycophenolate mofetil 2 g/day. Sirolimus and CsA were concentration controlled; trough levels of mycophenolic acid and prednisolone were also measured. RESULTS At 12 months, graft survival (92.5% sirolimus vs. 89.5% CsA), patient survival (97.5% sirolimus vs. 94.7% CsA), and the incidence of biopsy-proven acute rejection (27.5% sirolimus vs. 18.4% CsA) were not statistically different. The use of antibodies to treat suspected rejection episodes was also similar (7.5% sirolimus vs. 5.3% CsA). More sirolimus patients received bolus steroid therapy (20 vs. 11, P = 0.068). From month 2 onward, the calculated glomerular filtration rate was consistently higher in sirolimus-treated patients. The adverse events reported more frequently with sirolimus were thrombocytopenia (45% vs. 8%) and diarrhea (38% vs. 11%). In the CsA group, increased creatinine (18% vs. 39%), hyperuricemia (3% vs. 18%), cytomegalovirus infection (5% vs. 21%), and tremor (5% vs. 21%) were observed significantly more often. DISCUSSION Patient and graft survival and the incidence of biopsy-proven acute rejection at 12 months were comparable between sirolimus and CsA, whereas safety profiles were different. These data suggest that sirolimus may be used as primary therapy for the prevention of acute rejection.
Abnormally Phosphorylated Tau Protein in Senile Dementia of Lewy Body Type and Alzheimer Disease: Evidence that the Disorders Are Distinct

Summary The relationship between Alzheimer disease (AD) and dementia with Lewy bodies (senile dementia Lewy body type, or SDLT) and dementia in Parkinson's disease is unclear. AD pathology is characterised by both amyloid deposition and abnormal phosphorylation of tau in paired helical filaments (PHF-$tU). In AD, abnormally phosphorylated PHF-$tU is present in neurofibrillary tangles, in neuritic processes of senile plaques, and also in neuropil threads dispersed throughout the cerebral cortex. Cortical homogenates from 12 cases each of AD and SDLT, 13 cases of Parkinson's disease, and 11 normal controls were examined by Western blot analysis with antibodies that detect PHF-$tU. No PHF-$tU was found in Parkinson's disease or control cortex. No PHF-$tU was found in SDLT cases without histological evidence of tangles. PHF-$tU was detectable in SDLT cases with a low density of tangles, and large amounts of PHF-$tU were present in AD cases. This study demonstrates that abnormally phosphorylated PHF-$tU is only present where tangles are found and not in SDLT cases without tangles or with only occasional tangles. It is concluded that Lewy body dementias are distinct at a molecular level from AD.
Headache in Caucasian patients with Moyamoya angiopathy – a systematic cohort study

Background Headache is common in patients with Moyanoya angiopathy (MMA), but usually underestimated in its management and not well characterized. Methods A validated self-administered headache screening questionnaire and a telephone interview were used in order to investigate headache characteristics, frequency and pain intensity in a large cohort of 55 German patients with MMA. Results Thirty-seven patients (67.3%) had suffered from headache in the past year. Headache intensity was rated 3.2 ± 1.3 on a verbal rating scale from 0 to 10. Seventeen patients (47.9%) reported migraine-like headache, 10 patients (27.0%) reported tension type-like headache and 10 patients (27.0%) had a combination of both. The majority of patients with migraine-like headache (n = 10, 58.8%) described migrainous aura. Headache frequency and intensity improved significantly after revascularization surgery; however, nine patients developed new-onset headache postoperatively. Conclusion Headache is very common in MMA, often with a migraine-like phenotype. Tension type-like headache was also found in 27% of patients, which is a new finding that has not been reported before.
First Isolation of Bacteroides thetaiotaomicron from a Patient with a Cholesteatoma and Experiencing Meningitis

ABSTRACT A 45-year-old man with a cholesteatoma experienced purulent meningitis. Microbial analysis of cerebrospinal fluid yielded in pure culture a gram-negative bacillus. Phenotypic methods were suggestive of a Bacteroides distasonis or either a Bacteroides thetaiotaomicron or Bacteroides ovatus infection. The isolate was identified by 16S rRNA gene sequence analysis as B. thetaiotaomicron. This is the first case of B. thetaiotaomicron meningitis in pure culture.
Learning Disabilities and Central Auditory Dysfunction

Hearing loss, whether peripheral or central, compounds the communication and educational problems of the learning disabled student A central auditory processing disorder uniquely interferes with both the input and integration of verbal information, further resulting in a potentially permanent cognitive dysfunction during the developmental period of acquisition of language. Illustrative cases are presented that indicate the panorama of cognitive dysfunction associated with the learning disabled status. Methods of evaluation and identification and diagnostic criteria are correlated with auditory, visual, and academic performance. Comments regarding clinical awareness, prompt recognition, and ensuing individualized remediation are submitted.