Miracle-dz/UniFAR-FTUs huggingface dataset:
Evaluation of the Ca2+ Concentration in Purified Nerve Terminals: Relationship Between Ca2+ Homeostasis and Synaptosomal Preparation

Abstract: The presynaptic Ca2+ concentration ([Ca]i) was evaluated by studying intracellular free Ca2+ with quin‐2 and fura‐2 in synaptosomal preparations. The synaptosomal preparations were purified with hyperosmotic (sucrose) and isoosmotic (Percoll) density gradient centrifugation. Synaptosomes are most viable in the heavier fractions of the density gradients. These synaptosomal fractions exhibit the lowest [Ca]i, [204 ± 2 nM for Percoll (C‐band) synaptosomes, loaded at 30°C with the acetoxymethyl ester of fura‐2 (fura‐2‐AM)], a high stability during prolonged incubations at 37°C, and a more potent response to membrane depolarization by elevated extracellular [K+]. [Ca]i measurement was critically dependent on dye loading, calibration, type of dye used, synaptosomal preparation, and incubation temperature (30° or 37°C). Loading quin‐2 in synaptosomes inserts a considerable buffer component in the synaptosomal [Ca]i regulation, and consequently there is a quin‐2 dependency of [Ca]i, independent of endogenous heavy metal ions. Use of fura‐2 is preferable in synaptosomes, although above a critical fura2‐AM/protein ratio during loading ester hydrolysis is not complete, giving rise to errors in [Ca]i determination. Ionomycin is a selective tool to detect the presence of partially hydrolyzed esters and saturate indicators in the cytosol with Ca2+ for calibration. Parallel studies on lactate dehydrogenase and fura‐2 fluorescence indicate that synaptosomal viability is very sensitive to prolonged incubations at 37°C. This study shows the applicability of measuring steady‐state [Ca]i and dynamic [Ca]i changes quantitatively in fura‐2‐loaded synaptosomes. The possible involvement of different synaptosomal pools to explain the divergence in [Ca]i between different preparations and the interpretation in physiological terms of [Ca]i measured in synaptosomes are discussed.
Number needed to treat in fluconazole prophylaxis in the neonatal intensive care unit.

To the Editors: In a recent publication in this journal, Rueda et al conducted a study in a developing country and demonstrated that routine prophylactic use of fluconazole at birth versus placebo in 523 neonates less than 1250 g is associated with significant impact on the frequency of documented systemic Candida infection (CI). The control group developed CI in 21 (7.7%), whereas the treatment group had only 3 (1.1%) CI. The benefit of prophylactic fluconazole (PF) to prevent a systemic CI was represented by the number needed to treat (NNT) with a nominal value of 7 according to Rueda et al. This measure is calculated on the inverse of the absolute risk reduction. NNT of 7 means that it is necessary to treat 7 neonates with PF to prevent 1 CI. This NNT is calculated incorrectly. Considering the absolute risk reduction of 6.6% (7.7% 1.1%), the NNT should be 15.2 (calculated by 1/6.6%) instead of 7. Although this error does not change the statistical significance of the benefit of fluconazole in the study of Rueda et al (P 0.007), for pharmacoeconomic analysis studies, this error has an important impact. There is scant information from developing countries about PF in neonates, and the estimated NNT of 15.2 is acceptable. A broader view on the updated systematic review results of the Cochrane metaanalysis of 5 of 8 eligible trials on “systemic antifungal agents prophylatic in low birth weight infants,” issue 12, 2010, revealed that NNT for PF is in line with the confidence intervals of PF, with significant reduction in the risk of invasive fungal infection in the infants who received prophylaxis (NNT 11; 95% confidence interval, 7–33). Although it requires confirmation from future studies, the prophylactic use of fluconazole in premature infants of lower weight, with higher natural risk of systemic candidiasis (extremely low birth weight 1000 g), suggests a greater benefit: the calculated NNT is 2.68 for fungal colonization (22% PF 60% placebo) and NNT of 5 for systemic fungal infection (0% PF 20% placebo). Moreover, the systematic reviews still show a nonsignificant decrease in the risk of death before hospital discharge (typical relative risk, 0.74 95% confidence interval, 0.51–1.09 ). To date, the only untoward effects reported with fluconazole use in preterm infants are reversible elevations in hepatic enzymes or bilirubin, without sequelae.
Live demonstration: Feature extraction system using restricted Boltzmann machines on FPGA

Real-time results obtained from an unsupervised feature extraction system using Restricted Boltzmann Machines (RBMs) implemented on FPGA are presented. The feature extraction application is demonstrated using the MNIST dataset, and the weights storing features are visualized in real-time. A digit classification is also performed based on the learning results. Our demonstration system performs 134 times faster than the compared conventional CPU.
Life on a planet of its own: regulation of RNA polymerase I transcription in the nucleolus.

Mammalian cells contain 100 or more copies of tandemly repeated ribosomal RNA (rRNA) genes per haploid genome. These genes are transcribed with high efficiency to keep up with the cell’s metabolic activity and demand for ribosomes. Alterations in cell proliferation are accompanied by profound changes in the transcription rate of rRNA genes. Thus, by responding to changes in the cellular environment, transcription by RNA polymerase I (Pol I) ultimately determines ribosome production and the potential for cell growth and proliferation. There are several comprehensive reviews that discuss regulation of rRNA synthesis in vertebrates and yeast (Grummt 1999; Reeder 1999; Warner 1999; Moss and Stefanovsky 2002). However, new data have been produced even since the latest of these reviews that uncover the mechanisms that link Pol I transcription to cellular physiology. In this review, I restrict the background information to the minimal level that is required for understanding initiation complex formation at the rDNA promoter before proceeding to review the regulatory pathways that adapt cellular rRNA synthesis to cell metabolism and growth.
Mining Novel Candidate Imprinted Genes Using Genome-Wide Methylation Screening and Literature Review

Large-scale transcriptome and methylome data analyses obtained by high-throughput technologies have been enabling the identification of novel imprinted genes. We investigated genome-wide DNA methylation patterns in multiple human tissues, using a high-resolution microarray to uncover hemimethylated CpGs located in promoters overlapping CpG islands, aiming to identify novel candidate imprinted genes. Using our approach, we recovered ~30% of the known human imprinted genes, and a further 168 candidates were identified, 61 of which with at least three hemimethylated CpGs shared by more than two tissue types. Thirty-four of these candidate genes are members of the protocadherin cluster on 5q31.3; in mice, protocadherin genes have non-imprinted random monoallelic expression, which might also be the case in humans. Among the remaining 27 genes, ZNF331 was recently validated as an imprinted gene, and six of them have been reported as candidates, supporting our prediction. Five candidates (CCDC166, ARC, PLEC, TONSL, and VPS28) map to 8q24.3, and might constitute a novel imprinted cluster. Additionally, we performed a comprehensive compilation of known human and mice imprinted genes from literature and databases, and a comparison among high-throughput imprinting studies in humans. The screening for hemimethylated CpGs shared by multiple human tissues, together with the extensive review, appears to be a useful approach to reveal candidate imprinted genes.
Optimization, validation, and identification of two reliable antibodies for immunodetection of WNT5A

Abstract WNT5A is a secreted, noncanonical WNT signaling protein that has been reported to promote progression of several types of cancer, including oral squamous cell carcinoma. Many WNT5A antibodies are available commercially for immunohistochemistry (IHC) and western blot analysis. Validation of the primary antibodies, however, is often neglected. We characterized antibodies for detecting WNT5A by IHC and western blot analysis. We evaluated one polyclonal and three monoclonal commercially available WNT5A antibodies. After optimization of the IHC assay, all four antibodies showed cytoplasmic WNT5A expression in tissue samples; in contrast, only one antibody detected WNT5A in western blots. A pre-absorption test with recombinant WNT5A showed that AF645 and 3A4 antibodies specifically detected WNT5A in different assays. We suggest that the monoclonal 3A4 antibody is the most appropriate for use with IHC, while the polyclonal AF645 antibody is the best for western blot analysis.
Amelioration of impaired cerebral metabolism after severe acidotic ischemia by tirilazad posttreatment in dogs.

BACKGROUND AND PURPOSE Acidosis may contribute to ischemic injury by mobilizing iron because the iron chelator deferoxamine improves early metabolic recovery from hyperglycermic ischemia. Mobilized iron may then promote oxygen radical-induced lipid peroxidative injury during reperfusion. We tested the hypothesis that administration of the antioxidant tirilazad at the start of reperfusion improves early metabolic recovery after severe acidotic ischemia and ameliorates depletion of the endogenous antioxidant glutathione. METHODS In anesthetized dogs, arterial glucose concentration was increased to 500 to 600 mg/dL and global incomplete cerebral ischemia was produced for 30 minutes by ventricular fluid infusion to reduce perfusion pressure to 10 to 12 mm Hg. Metabolic recovery and intracellular pH were measured by phosphorus MR spectroscopy. In the first experiment, four groups of eight dogs each received either vehicle or 0.25, 1, or 2.5 mg/kg of tirilizad mesylate at reperfusion. Cerebral blood flow was measured with microspheres. In the second experiment, two groups of eight dogs each each received either vehicle or 2.5 mg/kg of tirilazad at reperfusion, and cortical glutathione was measured at 3 hours of reperfusion. RESULTS Cerebral blood flow decreased to approximately 6 mL/min per 100 g and intracellular pH decreased to approximately 5.6 during ischemia in all groups. In the vehicle group, ATP recovery was transient and pH remained less than 6.0. Cerebral blood flow, O2 consumption, and ATP eventually declined to near-zero levels by 3 hours. Recovery was improved by tirilazad posttreatment in a dose-dependent fashion. At the highest dose, cerebral blood flow and O2 consumption were sustained near preischemic levels, and five of eight dogs had recovery of ATP greater than 50% and of pH greater than 6.7. Recovery of ATP and phosphocreatine became significantly greater than that in the vehicle group by 17 minutes of reperfusion despite similar levels of early hyperemia, indicating that the drug was acting before the onset of hypoperfusion. Cortical glutathione concentration in the vehicle group was 27% less than that in the tirilazad group and 34% less than that in nonischemic controls. CONCLUSIONS Decreased depletion of the endogenous antioxidant glutathione is consistent with tirilazad acting as an antioxidant in vivo. Improvement in high-energy phosphate recovery 17 minutes after starting tirilazad infusion during reperfusion is consistent with an early onset of a functionally significant oxygen radical injury. Thus, severe acidosis appears to contribute to early ischemic injury through an oxygen radical mechanism sufficient to impede metabolic recovery.
The Effects of Frequency of Social Interaction and Relationship Closeness on Reward Allocation

Abstract In this study of how the frequency of interaction affects decisions on reward allocation, 320 Chinese adults were randomly assigned to 8 conditions and instructed to read a scenario describing 2 co-workers completing a task and obtaining a bonus. The 8 conditions differed in the type of relationships, the frequency of past interaction between co-workers, and the possibility of future interaction. Each participant was instructed to act as the allocator and to hypothetically allocate a monetary reward to the co-worker. The results indicated that both past interaction and future interaction between the co-workers significantly influenced the allocation decision. These results also suggested that the frequency of 2 individuals' past interaction can be predictive of the closeness of their relationship.
Treatment of borderline personality disorder with risperidone.

BACKGROUND Of the various Axis II disorders, borderline personality disorder (BPD) is among the more critical to treat. There are at present few results in terms of clinical outcome with the psychotropic agents available. Possible targets for pharmacotherapy are affective symptoms, cognitive disturbances, and impulsive, self-injurious behaviors. In previous studies, atypical antipsychotics at low-to-moderate doses provided symptom reduction with good tolerability. Our purpose was to assess the efficacy of risperidone in BPD, focusing on its effects on impulsive-aggressive behavior. METHOD Fifteen BPD outpatients (DSM-IV diagnosis) with prominent histories of aggressive behavior were included in an 8-week open-label study with risperidone at low-to-moderate doses. Axis II codiagnoses included antisocial personality disorder (N = 4). Exclusion criteria included current Axis I diagnosis or any major medical or neurologic illness. Efficacy measures were the 21-item Hamilton Rating Scale for Depression, the Brief Psychiatric Rating Scale, the DSM-IV Global Assessment of Functioning, and the self-rated Aggression Questionnaire. Evaluations were carried out at baseline and at the end of the treatment. RESULTS Thirteen patients completed the trial; 2 patients dropped out because of lack of compliance. Final mean dose of risperidone was 3.27 mg/day. There was a significant (p = .0057) reduction in aggression based on Aggression Questionnaire scores. This amelioration was coupled with an overall improvement, including a reduction in depressive symptoms and an increase in energy and global functioning. CONCLUSION Risperidone at low-to-moderate doses can improve BPD symptomatology. Further studies are needed to explore the efficacy of risperidone versus placebo as well as in comparison to other potential treatments for BPD.
Recombinant interferon gamma augments phagocyte superoxide production and X-chronic granulomatous disease gene expression in X-linked variant chronic granulomatous disease.

We examined the potential of interferon gamma (IFN-gamma) to ameliorate the physiologic defect of chronic granulomatous disease (CGD) by studying its effects on CGD phagocyte superoxide generation, NADPH oxidase kinetics, cytochrome b559 content, and expression of X-CGD (the gene for the X-linked disease). Granulocytes and macrophages from three patients in two kindreds with "variant" X-linked CGD (i.e., with very low, but detectable, baseline superoxide-generating activity) responded to IFN-gamma with enhanced nitroblue tetrazolium reduction and two- to eightfold increases in superoxide generation. IFN-gamma did not augment the respiratory burst activity of phagocytes from patients with "classic" CGD (i.e., no detectable baseline superoxide generation) or autosomal variant CGD. Incubation of a responding patient's granulocytes with IFN-gamma nearly doubled the maximal velocity for the NADPH oxidase, but did not change its abnormal Michaelis constant. Although the interferon-treated CGD granulocytes produced superoxide at a rate 40% of normal, the cytochrome b spectrum remained undetectable. IFN-gamma treatment of cultured monocytes from an IFN-gamma-responsive CGD patient increased the steady state level of RNA transcripts from the X-CGD gene from barely detectable up to approximately 5% of normal.
LMNA cardiomyopathy: cell biology and genetics meet clinical medicine

Mutations in the LMNA gene, which encodes A-type nuclear lamins (intermediate filament proteins expressed in most differentiated somatic cells), cause a diverse range of diseases, called laminopathies, that selectively affect different tissues and organ systems. The most prevalent laminopathy is cardiomyopathy with or without different types of skeletal muscular dystrophy. LMNA cardiomyopathy has an aggressive clinical course with higher rates of deadly arrhythmias and heart failure than most other heart diseases. As awareness among physicians increases, and advances in DNA sequencing methods make the genetic diagnosis of LMNA cardiomyopathy more common, cardiologists are being faced with difficult questions regarding patient management. These questions concern the optimal use of intracardiac cardioverter defibrillators to prevent sudden death from arrhythmias, and medical interventions to prevent heart damage and ameliorate heart failure symptoms. Data from a mouse model of LMNA cardiomyopathy suggest that inhibitors of mitogen-activated protein kinase (MAPK) signaling pathways are beneficial in preventing and treating cardiac dysfunction; this basic research discovery needs to be translated to human patients.
Empirical Study on Sustainable Opportunities Recognition. A Polyvinyl Chloride (PVC) Joinery Industry Analysis Using Augmented Sustainable Development Process Model

This paper analyzes factors influencing recognition of sustainable opportunities by using an augmented sustainability process model. The conceptual model used two main factors, Knowledge and Motivation, and one moderating variable, Social embeddedness. We investigated entrepreneurs from PVC joinery industry and concluded that while market orientation and sustainable entrepreneurial orientation definitely and positively influence sustainable opportunity recognition, others variables like knowledge of the natural/communal environment, awareness of sustainable development or focus on success have less support. Among all variables analyzed, perception of the threat of the natural/communal environment and altruism toward others have the poorest impact on opportunity recognition. Finally, we concluded that social embeddedness has a moderating effect on sustainable opportunity recognition, even though the results were mixed.
Presymptomatic studies in ALS

ALS= : amyotrophic lateral sclerosis; DTI= : diffusion tensor imaging; fALS= : familial amyotrophic lateral sclerosis; FTD= : frontotemporal dementia; LMN= : lower motor neuron; MN= : motor neuron; MRS= : magnetic resonance spectroscopy; MUNE= : motor unit number estimation; Pre-fALS= : Pre-symptomatic Familial ALS study; UMN= : upper motor neuron The last 15 years have witnessed a significant shift in our understanding of neurodegenerative diseases. We increasingly recognize that the degenerative process in disorders such as Alzheimer disease,1 Huntington disease,2 and Parkinson disease3,–,5 begins years, if not decades, prior to the appearance of typical clinical manifestations. Whether ALS is also characterized by a prolonged presymptomatic period is unclear, but definitively answering this question is likely to have profound implications for understanding disease biology, uncovering environmental risk factors, developing effective therapies, and even disease prevention. To effectively study presymptomatic ALS, one must first identify people who are at risk for developing disease. Although advancing age is a risk factor for ALS, the broad range of age at symptom onset limits its utility in identifying at-risk individuals. Absent any known environmental risk factor, it is currently only possible to study individuals genetically predisposed to developing ALS—namely, presymptomatic gene mutation carriers. We begin this review with …
On the topological aspects of the theory of represented spaces

Represented spaces form the general setting for the study of computability derived from Turing machines. As such, they are the basic entities for endeavors such as computable analysis or computable measure theory. The theory of represented spaces is well-known to exhibit a strong topological flavour. We present an abstract and very succinct introduction to the field; drawing heavily on prior work by Escardo, Schroder, and others. Central aspects of the theory are function spaces and various spaces of subsets derived from other represented spaces, and - closely linked to these - properties of represented spaces such as compactness, overtness and separation principles. Both the derived spaces and the properties are introduced by demanding the computability of certain mappings, and it is demonstrated that typically various interesting mappings induce the same property.
Effect of recent alcohol intake on parathyroid hormone and mineral metabolism in men.

The mechanisms by which alcohol intake, particularly moderate alcohol intake, effects bone metabolism are poorly defined. We have examined the relationship between mineral metabolism and recent self-reported alcohol intake (SRAI) across a wide range of such intakes in a series of 104 men aged 32 to 78 years of age in an outpatient setting. A morning nonfasting urine, serum specimen and recent SRAI were obtained from each subject. SRAI was reported as between 0 and 45 oz/week. SRAI correlated positively with liver function tests, including serum bilirubin (r = 0.30, p = 0.002), alkaline phosphatase (r = 0.30, p = 0.004), and aspartate aminotransferase (SGOT) (r = 0.29, p = 0.006). SRAI correlated with serum calcium corrected for albumin (r = -0.39, p < 0.001), estradiol (r = 0.43, p < 0.001), and immunoreactive parathyroid hormone (iPTH) (r = -0.51, p < 0.001), as well as urinary calcium (per 100 mg of creatinine) (r = 0.55, p < 0.001). We have arbitrarily divided the participants into two groups on the basis of their reported alcohol intake. Individuals in the first group had intakes ranging from none to moderate intake (drank 8.4 oz or less of ethanol per week, equivalent to an average of two drinks daily or less). Those in the second group had moderate or heavier intake, with >8.4 oz of ethanol intake/week. Mean serum iPTH was significantly greater in those in the first group (none to moderate), compared with the second group (moderate or heavier) (56.0 +/- 3.4 and 39.9 +/- 2.0 pM/liter, respectively). Calcium corrected for serum albumin was significantly greater in individuals in the first, compared with the second, group (9.23 +/- 0.05 vs. 8.88 +/- 0.07 mg/dl, respectively). In addition, urinary calcium (corrected per 100 mg of creatinine) was significantly lower in the former, compared with the latter (3.1 +/- 0.4 vs. 8.4 +/- 1.1 mg/100 mg of creatinine, respectively). Similarly, urinary excretion of collagen crosslinks (corrected per 100 mg of creatinine) was significantly less in men in the second group, compared with the first group (316 +/- 38 vs. 530 +/- 78 nM/100 mg of creatinine, respectively). Not surprisingly, a series of correlations between iPTH and age, 250-hydroxyvitamin D, and testosterone were significant in individuals with none to moderate SRAI, but not moderate or heavier SRAI. Significant independent predictors of serum iPTH in the entire group of men were age (beta = 0.215, p = 0.025), SRAI (beta = -0.281, p = 0.003), 250-hydroxyvitamin D (beta = -0.309, p = 0.002), and testosterone (beta = -184, p = 0.048). We have concluded that, in free-living men, alcohol intake >8.4 oz/week was associated with decreased serum iPTH concentrations.