Miracle-dz/UniFAR-FTUs huggingface dataset:
Activation and desensitization of N‐methyl‐D‐aspartate receptors in nucleated outside‐out patches from mouse neurones.

1. Activation and desensitization of N‐methyl‐D‐aspartate (NMDA) receptors were studied in large outside‐out patches excised from cultured embryonic neurones dissociated from mouse forebrain. The patches were exposed to rapid changes of NMDA or L‐glutamate concentrations in the presence of glycine at concentrations (10‐20 microM) saturating the modulatory site of the NMDA receptor. 2. Immediately after formation of the patch the responses to NMDA and L‐glutamate showed a slow and small desensitization, even with high concentrations of agonist. During the following hour, the peak response either decreased or remained relatively stable, but in all cases the desensitization increased and accelerated until it stabilized. In this 'stabilized’ state, the desensitization produced by high concentrations of NMDA (1 mM) or L‐glutamate (300 microM) had an exponential time course, with a time constant of about 30 ms. The ratio of the peak over the steady‐state current was in the order of 40 for NMDA and about 30 for L‐glutamate. 3. Concentration‐response curves were built for the peak and the plateau responses, for NMDA and for L‐glutamate. The comparison of these curves indicated that (i) the EC50 of the peak (K(app) was always higher than the EC50 of the plateau (Kss); (ii) the two EC50 values for NMDA (K(app) and Kss) were higher than those for L‐glutamate; (iii) the Hill coefficient was close to 1.4 for each of the four curves. 4. The application of NMDA or L‐glutamate at a low concentration for 3 s periods reduced the response to a subsequent application of the same agonist at a saturating concentration. The IC50 of this ‘predesensitization’, termed Kpre, was lower than the EC50 of the steady‐state response, Kss. 5. The onset rates of desensitization increased with the concentration of agonist. The EC50 of this relation was close to the value of K(app). 6. The decay of the currents at the end of a 3 s application of agonist was usually well described by the sum of two exponentials both of which were faster for NMDA than for L‐glutamate. 7. The recovery from desensitization after a long (3 s) pulse of agonist was approximately exponential, with a time constant of about 0.5 s for NMDA and about 3.5 s for L‐glutamate. (ABSTRACT TRUNCATED AT 400 WORDS)
Real-Time Detection of Noroviruses in Surface Water by Use of a Broadly Reactive Nucleic Acid Sequence-Based Amplification Assay

ABSTRACT Noroviruses are the most common agents causing outbreaks of viral gastroenteritis. Outbreaks originating from contaminated drinking water and from recreational waters have been described. Due to a lack of cell culture systems, noroviruses are detected mostly by molecular methods. Molecular detection assays for viruses in water are often repressed by inhibitory factors present in the environment, like humic acids and heavy metals. To study the effect of environmental inhibitors on the performance of nucleic acid sequence-based amplification (NASBA), we developed a real-time norovirus NASBA targeting part of the RNA-dependent RNA polymerase (RdRp) gene. Specificity of the assay was studied with 33 divergent clones that contained part of the targeted RdRp gene of noroviruses from 15 different genogroups. Viral RNA originated from commercial oysters, surface waters, and sewage treatment plants in The Netherlands. Ninety-seven percent of the clones derived from human noroviruses were detected by real-time NASBA. Two clones containing animal noroviruses were not detected by NASBA. We compared the norovirus detection by real-time NASBA with that by conventional reverse transcriptase PCR (RT-PCR) with large-volume river water samples and found that inhibitory factors of RT-PCR had little or no effect on the performance of the norovirus NASBA. This consequently resulted in a higher sensitivity of the NASBA assay than of the RT-PCR. We show that by combining an efficient RNA extraction method with real-time NASBA the sensitivity of norovirus detection in water samples increased at least 100 times, which consequently has implications for the outcome of the infectious risk assessment.
Genetic Dissection of the Vav2-Rac1 Signaling Axis in Vascular Smooth Muscle Cells

ABSTRACT Vascular smooth muscle cells (vSMCs) are key in the regulation of blood pressure and the engagement of vascular pathologies, such as hypertension, arterial remodeling, and neointima formation. The role of the Rac1 GTPase in these cells remains poorly characterized. To clarify this issue, we have utilized genetically engineered mice to manipulate the signaling output of Rac1 in these cells at will using inducible, Cre-loxP-mediated DNA recombination techniques. Here, we show that the expression of an active version of the Rac1 activator Vav2 exclusively in vSMCs leads to hypotension as well as the elimination of the hypertension induced by the systemic loss of wild-type Vav2. Conversely, the specific depletion of Rac1 in vSMCs causes defective nitric oxide vasodilation responses and hypertension. Rac1, but not Vav2, also is important for neointima formation but not for hypertension-driven vascular remodeling. These animals also have allowed us to dismiss etiological connections between hypertension and metabolic disease and, most importantly, identify pathophysiological programs that cooperate in the development and consolidation of hypertensive states caused by local vascular tone dysfunctions. Finally, our results suggest that the therapeutic inhibition of Rac1 will be associated with extensive cardiovascular system-related side effects and identify pharmacological avenues to circumvent them.
Two Cyclic Dipeptides from Pseudomonas fluorescens GcM 5-1 A Carried by the Pine Wood Nematode and Their Toxicities to Japanese Black Pine Suspension Cells and Seedlings in vitro

Pseudomonas fluorescens GcM5-1A, isolated from the pine wood nematode (PWN), Bursaphelenchus xylophilus, was cultured in Luria Broth medium (LB). The clarified culture was extracted with ethyl acetate, and two dipeptides were purified from the extract. The chemical structures of 1 and 2 were identified as cyclo(-Pro-Val-)and cyclo(-Pro-Tyr-), respectively, by MS, H NMR, C NMR, H-H COSY, H -C COSY spectra. Bioassay results showed that the two compounds were toxic to both suspension cells and seedlings of Pinus thunbergii, which may offer some clues to research the mechanism of pine wilt disease caused by PWN.
Echocardiographic measurements in greyhounds, whippets and Italian greyhounds--dogs with a similar conformation but different size.

OBJECTIVE To determine the effect of body size on various echocardiographic measurements in dogs of widely differing size, but identical body conformation. DESIGN A randomised echocardiographic study of healthy sighthounds. ANIMALS 60 dogs comprising an equal number (20) of racing Greyhounds, Whippets and Italian Greyhounds. PROCEDURE Following sedation with acepromazine and morphine, and acclimatization, a thorough echocardiographic examination was performed on each dog using standard methods. RESULTS Dimensions measured echocardiographically were highly correlated with body size. These data were subsequently examined using analyses of variance and regression. Body surface area was the best overall predictor of dimensional measurements. In comparison to previous studies using dogs of differing size and conformation, the spread of values for measurements plotted against body surface area showed substantially narrower ranges. Thus, the relationship between echocardiographic measurements and body surface area was much closer for dogs with an identical somatotype than for dogs of differing size and conformation. Commonly used ejection phase indices (fractional shortening, ejection fraction and velocity of circumferential fibre shortening) were negatively correlated with body size. In contrast, the thickening fraction of the left ventricular posterior wall, another ejection phase index, was independent of body weight and body surface area for all three breeds and when the data were pooled. CONCLUSION Taken in consideration with previous work, this study demonstrates that body conformation and body size both influence canine echocardiographic measurements. Commonly used ejection phase indices are significantly affected by body size, with larger sighthounds having lower values. A more appropriate method of quantitating left ventricular function may be the determination of the thickening fraction of the left ventricular posterior wall.
Weight Regain: No Link to Success in a Real-Life Multiday Boxing Tournament.

PURPOSE Combat-sport athletes acutely reduce body mass (BM) before weigh-in in an attempt to gain a size/strength advantage over smaller opponents. Few studies have investigated these practices among boxers and none have explored the impact of this practice on competitive success. METHODS One hundred (30 women, 70 men) elite boxers participating in the Australian national championships were weighed at the official weigh-in and 1 h before each competition bout. Regain in BM after weigh-in was compared between finalists and nonfinalists, winners and losers of each fight, men and women, and weight divisions. Boxers were surveyed on their pre- and post-weigh-in nutrition practices. RESULTS The lightest men's weight category displayed significantly greater relative BM regain than all other divisions, with no difference between other divisions. BM prebout was higher than official weigh-in for men (2.12% ± 1.62%; P < .001; ES = 0.13) and women (1.49% ± 1.65%; P < .001; ES = 0.11). No differences in BM regain were found between finalists and nonfinalists, winners and losers of individual bouts, or between preliminary or final bouts. BM regain was significantly greater (0.37% BM, P < .001; ES = 0.25) before an afternoon bout compared with a morning bout. CONCLUSIONS Boxers engage in acute BM-loss practices before the official competition weigh-in, but this does not appear to affect competition outcomes, at least when weight regain between weigh-in and fighting is used as a proxy for the magnitude of acute loss. While boxers recognize the importance of recovering after weigh-in, current practice is not aligned with best-practice guidance.
Sequelae of Arteriosclerosis of the Aorta and Coronary Arteries: A Statistical Study in Diabetes Mellitus

The relation of diabetes mellitus to arteriosclerosis has recently been summed up by Edwards as follows: "Diabetes is commonly recognized as having a positive influence in coronary atherosclerosis. It is a relatively common experience to perform necropsies on diabetic persons who die of severe atherosclerosis at ages much younger than the average. " In the first edition of their book, Pathology of Diabetes Mellitus, Warren and LeCompte stated that they had not seen an autopsy on a diabetic patient whose disease had lasted five years or more, free from arteriosclerosis, regardless of age. However, they subsequently encountered a few cases with duration over five years practically free from arteriosclerosis. The evaluation of the severity of arteriosclerosis appears to be, in large part, a subjective one. It would seem that a more objective approach to the problem would be an evaluation of the frequency of sequelae of arteriosclerosis. For example, White has pointed out that diabetic patients with hypercholesterolemia are fifteen times more likely to develop arteriosclerosis than diabetic patients with normal cholesterol levels, but it has not been established that patients with controlled diabetes have any striking aberrations of the serum cholesterol. *" Despite such considerations it is generally accepted that the positive effect of diabetes on arteriosclerosis is the result of increased serum lipid levels associated with this disease. Strangely, when one evaluates the frequency of sequelae of arteriosclerosis they are peculiarly limited to the heart and extremities, and Warren and LeCompte have pointed out that cerebral arteriosclerosis is no more frequently a cause of death in diabetic patients than in nondiabetics. Furthermore, Blumenthal and coworkers" have been unable to demonstrate, on a microscopic level, an increased intensity of
Neural Mechanisms of Visual Attention: How Top-Down Feedback Highlights Relevant Locations

Attention helps us process potentially important objects by selectively increasing the activity of sensory neurons that represent the relevant locations and features of our environment. This selection process requires top-down feedback about what is important in our environment. We investigated how parietal cortical output influences neural activity in early sensory areas. Neural recordings were made simultaneously from the posterior parietal cortex and an earlier area in the visual pathway, the medial temporal area, of macaques performing a visual matching task. When the monkey selectively attended to a location, the timing of activities in the two regions became synchronized, with the parietal cortex leading the medial temporal area. Parietal neurons may thus selectively increase activity in earlier sensory areas to enable focused spatial attention.
Takotsubo cardiomyopathy: pathophysiology and treatment

Since it was first described 20 years ago in Japan, takotsubo cardiomyopathy has received considerable interest from the medical community around the world, particularly in recent years. Although takotsubo cardiomyopathy was originally described as a transient, stress-induced dysfunction of the left ventricular apex, other morphological subtypes have now been described. The pathogenesis of this disorder is likely to be catecholamine mediated myocyte damage and microvascular dysfunction; however, a number of possible alternative theories have been suggested. These include oxidative stress, transient coronary obstruction and oestrogen deficiency, the last explaining the high prevalence of takotsubo cardiomyopathy in women. The treatment remains largely supportive; however, a number of agents have been implicated in the acute stage and long term. Although most of the patients show complete recovery, there is a high risk of complications at the initial presentation requiring intense support.
Structured Outpatient Peritoneal Dialysis Catheter Insertion Is Safe and Cost-Saving

♦ Background: Data about outcomes and costs for peritoneal catheter insertion on an outpatient basis are scarce. ♦ Methods: Using patient files, all peritoneal dialysis (PD) catheter insertions performed between 2004 and 2009 in a single-center tertiary care institution for adult patients were located. Patient demographics, complications, hospitalizations, survival, and treatment modality changes were recorded. Procedure-related expenses were valued as actual production costs. ♦ Results: During the study period, 106 PD catheters were inserted. In 46 cases, the patients were admitted electively for catheter insertion; 19 catheters were placed during admission for other medical reasons; and 41 catheters were placed on an outpatient basis. Among the study patients (54.7 ± 16.0 years of age), 45% were diabetic. Early (<30 days) catheter-related complications occurred in 22% of patients. The incidences of technique failure and any complication within 90 days were 10% and 38% respectively. The occurrence of complications was not statistically significantly different for outpatients and electively admitted patients. Average costs for catheter insertion were higher in electively hospitalized patients than in outpatients (€2320 ± €960 vs €1346 ± €208, p < 0.000). ♦ Conclusions: Compared with an inpatient procedure, outpatient insertion of a PD catheter results in similar outcomes at a lower cost.
β‐Catenin—A supporting role in the skeleton

In the last 5 years a role for β‐catenin in the skeleton has been cemented. Beginning with mutations in the Lrp5 receptor that control β‐catenin canonical downstream signals, and progressing to transgenic models with bone‐specific alteration of β‐catenin, research has shown that β‐catenin is required for normal bone development. A cell critical to bone in which β‐catenin activity determines function is the marrow‐derived mesenchymal stem cell (MSC), where sustained β‐catenin prevents its distribution into adipogenic lineage. β‐Catenin actions are less well understood in mature osteoblasts: while β‐catenin contributes to control of osteoclastic bone resorption via alteration of the osteoprotegerin/RANKL ratio, a specific regulatory role during osteoblast bone synthesis has not yet been determined. The proven ability of mechanical factors to prevent β‐catenin degradation and induce nuclear translocation through Lrp‐independent mechanisms suggests processes by which exercise might modulate bone mass via control of lineage allocation, in particular, by preventing precursor distribution into the adipocyte pool. Effects resulting from mechanical activation of β‐catenin in mature osteoblasts and osteocytes likely modulate bone resorption, but whether β‐catenin is involved in osteoblast synthetic function remains to be proven for both mechanical and soluble mediators. As β‐catenin appears to support the downstream effects of multiple osteogenic factors, studies clarifying when and where β‐catenin effects occur will be relevant for translational approaches aimed at preventing bone loss and terminal adipogenic conversion. J. Cell. Biochem. 110: 545–553, 2010. © 2010 Wiley‐Liss, Inc.
The Role of C-terminal Tyrosine Phosphorylation in the Regulation of SHP-1 Explored via Expressed Protein Ligation*

The protein-tyrosine phosphatase SHP-1 plays a variety of roles in the “negative” regulation of cell signaling. The molecular basis for the regulation of SHP-1 is incompletely understood. Whereas SHP-1 has previously been shown to be phosphorylated on two tail tyrosine residues (Tyr536and Tyr564) by several protein-tyrosine kinases, the effects of these phosphorylation events have been difficult to address because of the intrinsic instability of the linkages within a protein-tyrosine phosphatase. Using expressed protein ligation, we have generated semisynthetic SHP-1 proteins containing phosphotyrosine mimetics at the Tyr536 and Tyr564 sites. Two phosphonate analogues were installed, phosphonomethylenephenylalanine (Pmp) and difluorophosphonomethylenephenylalanine (F2Pmp). Incorporation of Pmp at the 536 site led to 4-fold stimulation of the SHP-1 tyrosine phosphatase activity whereas incorporation at the 564 site led to no effect. Incorporation of F2Pmp at the 536 site led to 8-fold stimulation of the SHP-1 tyrosine phosphatase activity and 1.6-fold at the 564 site. A combination of size exclusion chromatography, phosphotyrosine peptide stimulation studies, and site-directed mutagenesis led to the structural model in which tyrosine phosphorylation at the 536 site engages the N-Src homology 2 domain in an intramolecular fashion relieving basal inhibition. In contrast, tyrosine phosphorylation at the 564 site has the potential to engage the C-Src homology 2 domain intramolecularly, which can modestly and indirectly influence catalytic activity. The finding that phosphonate modification at each of the 536 and 564 sites can promote interaction with the Grb2 adaptor protein indicates that the intramolecular interactions fostered by post-translational modifications of tyrosine are not energetically strong and susceptible to intermolecular competition.
The structural basis of cell-mediated immunological reactions of collagen. Recognition by the cutaneous delayed hypersensitivity reaction in guinea-pigs of conformational alterations of rat and calf skin collagen.

Guinea-pigs were sensitized for delayed cutaneous hypersensitivity reactions to native and thermally denatured rat and calf skin collagens. Native and denatured collagens sensitized the animals to a comparable degree. However, the immune mechanism of the animals differentiated the purely conformational alterations of the molecule, even across the species barrier. Denaturation was accompanied by loss of immunogenic information as well as by exposure of new determinants, not recognizable in the native molecule. The sera of these animals were examined for the presence of antibodies reactive with native collagen in a passive haemagglutination system. Low levels of such antibodies were found in animals sensitized with native collagen and no antibodies reactive with native collagen in animals sensitized with denatured molecules although the latter group of guinea-pigs responded well with delayed skin reactions to native collagen.
Bacterial pathogens modulate an apoptosis differentiation program in human neutrophils

Human polymorphonuclear leukocytes (PMNs or neutrophils) are essential to the innate immune response against bacterial pathogens. Recent evidence suggests that PMN apoptosis facilitates resolution of inflammation during bacterial infection. Although progress has been made toward understanding apoptosis in neutrophils, very little is known about transcriptional regulation of this process during bacterial infection. To gain insight into the molecular processes that facilitate resolution of infection, we measured global changes in PMN gene expression during phagocytosis of a diverse group of bacterial pathogens. Genes encoding key effectors of apoptosis were up-regulated, and receptors critical to innate immune function were down-regulated during apoptosis induced by phagocytosis of Burkholderia cepacia, Borrelia hermsii, Listeria monocytogenes, Staphylococcus aureus, and Streptococcus pyogenes. Importantly, we identified genes that comprise a common apoptosis differentiation program in human PMNs after phagocytosis of pathogenic bacteria. Unexpectedly, phagocytosis of Str. pyogenes induced changes in neutrophil gene expression not observed with other pathogens tested, including down-regulation of 21 genes involved in responses to IFN. Compared with other bacteria, PMN apoptosis was significantly accelerated by Str. pyogenes and was followed by necrosis. Thus, we hypothesize that there are two fundamental outcomes for the interaction of bacterial pathogens with neutrophils: (i) phagocytosis of bacteria induces an apoptosis differentiation program in human PMNs that contributes to resolution of bacterial infection, or (ii) phagocytosis of microorganisms such as Str. pyogenes alters the apoptosis differentiation program in neutrophils, resulting in pathogen survival and disease.