Miracle-dz/UniFAR-FTUs huggingface dataset:
Kinetic validation of 6‐NBDG as a probe for the glucose transporter GLUT1 in astrocytes

In recent years, the use of fluorescent glucose analogs has allowed the study of rapid transport modulation in heterogeneous cell cultures and complex tissues. However, the kinetic behavior of these tracers is not conventional. For instance, the fluorescent glucose analog 6‐NBDG permeates the cell 50–100 times slower than glucose but the uptake of 6‐NBDG is almost insensitive to glucose, an observation that casts doubts as to the specificity of the uptake pathway. To investigate this apparent anomaly in cultured astrocytes, which are rich in the glucose transporter GLUT1, we first estimated the kinetic parameters of 6‐NBDG uptake, which were then incorporated into the kinetic model of GLUT1. The main outcome of the analysis was that 6‐NBDG binds to GLUT1 with 300 times higher affinity than glucose, which explains why its uptake is not efficiently displaced by glucose. The high binding affinity of 6‐NBDG also explains why cytochalasin B is less effective at inhibiting 6‐NBDG uptake than at inhibiting glucose uptake. We conclude that 6‐NBDG, used at low concentrations, permeates into astrocytes chiefly through GLUT1, and advise that the exofacial GLUT1 inhibitor 4,6‐ethylidine‐d‐glucose be used, instead of glucose, as the tool of choice to confirm the specificity of 6‐NBDG uptake.
Acetate transport and utilization in the rat brain

Acetate, a glial‐specific substrate, is an attractive alternative to glucose for the study of neuronal‐glial interactions. The present study investigates the kinetics of acetate uptake and utilization in the rat brain in vivo during infusion of [2‐13C]acetate using NMR spectroscopy. When plasma acetate concentration was increased, the rate of brain acetate utilization (CMRace) increased progressively and reached close to saturation for plasma acetate concentration > 2–3 mM, whereas brain acetate concentration continued to increase. The Michaelis–Menten constant for brain acetate utilization ( = 0.01 ± 0.14 mM) was much smaller than for acetate transport through the blood–brain barrier (BBB) ( = 4.18 ± 0.83 mM). The maximum transport capacity of acetate through the BBB ( = 0.96 ± 0.18 μmol/g/min) was nearly twofold higher than the maximum rate of brain acetate utilization ( = 0.50 ± 0.08 μmol/g/min). We conclude that, under our experimental conditions, brain acetate utilization is saturated when plasma acetate concentrations increase above 2–3 mM. At such high plasma acetate concentration, the rate‐limiting step for glial acetate metabolism is not the BBB, but occurs after entry of acetate into the brain.
Supramolecular elastomer based on polydimethylsiloxanes (SESi) film: synthesis, characterization, biocompatibility, and its application in the context of wound dressing

Supramolecular elastomer based on polydimethylsiloxanes (SESi) is a kind of novel elastomer cross-linked by the multihydrogen bonds supplied by the functional groups linked to the end of the PDMS chains, such as amide, imidazolidone, pending urea (1,1-dialkyl urea), and bridging urea (1,3-dialkyl urea). SESi showed lower glass transition temperature (T g) at about −113 °C because of the softer chain of PDMS, and could show real rubber-like elastic behaviors and acceptable water vapor transmission rate under room temperature. The high biocompatibility of SESi in the form of films was demonstrated by the cytotoxicity evaluation (MTT cytotoxicity assay and direct contact assay), hemolysis assay, and skin irritation evaluation. Based on detailed comparisons between commercial Tegaderm™ film and SESi film using a full-thickness rat skin model experiment, it was found that SESi film showed similar wound contraction rate as that of Tegaderm™ film on day seven, 10, and 14; only on day five, SESi film showed a significant (p < 0.05) lower wound contraction rate. And, the wounds covered with SESi film were filled with new epithelium without any significant adverse reactions, similar with that of Tegaderm™ film.
The Meaning of Letting Go: The Lived Experience for Caregivers of Persons at the End of Life

Parse’s research method was used to describe the structure of the experience of letting go as portrayed by caregivers of persons at the end of life. Eight female caregivers told their stories of what it means to let go of their loved one through the dying process. The participants were asked to bring to the discussion a photograph that most represented the meaning of letting go for them. All but two participants brought a photograph of the participant physically holding their loved one, an image that supports the paradoxical nature of letting go. For these women, the structure of the meaning of letting go was “persevering in presence despite hardships while bearing witness to wavering decline life unfolds with a new view. ”
Neuronal glucose but not lactate utilization is positively correlated with NMDA‐induced neurotransmission and fluctuations in cytosolic Ca2+ levels

Although the brain utilizes glucose for energy production, individual brain cells may to some extent utilize substrates derived from glucose. Thus, it has been suggested that neurons consume extracellular lactate during synaptic activity. However, the precise role of lactate for fuelling neuronal activity is still poorly understood. Recently, we demonstrated that glucose metabolism is up‐regulated in cultured glutamatergic neurons during neurotransmission whereas that of lactate is not. Here, we show that utilization of glucose but not lactate correlates with NMDA‐induced neurotransmitter glutamate release in cultured cerebellar neurons from mice. Pulses of NMDA at 30, 100, and 300 μM, leading to a progressive increase in both cytosolic [Ca2+] and release of glutamate, increased uptake and metabolism of glucose but not that of lactate as evidenced by mass spectrometric measurement of 13C incorporation into intracellular glutamate. In this manuscript, a cascade of events for the preferential neuronal utilization of glucose during neurotransmission is suggested and discussed in relation to our current understanding of neuronal energy metabolism.
Sialic acid Levels in smokers and nonsmokers with malignant pleural effusion

Objective: The present study is designed to measure sialic acid in pleural effusion and in serum samples of patients to differentiate between malignant and nonmalignant diseases. Materials and Methods: The present study was conducted on 30 patients with malignant pleural effusion and 30 patients with nonmalignant pleural effusion. Pleural fluid and blood samples were taken at the time of admission, before starting any treatment. Sialic acid levels were estimated in serum and pleural fluid by Warren′s Thiobarbituric Acid (TBA) method. Results: In the present study, serum sialic acid levels were higher in group II as compared to group I. In the present study, pleural fluid (PF) sialic acid levels and PF/serum(S) ratio was higher in malignant pleural effusion (though difference was not statistically significant). Smokers in group II had higher serum sialic acid as compared to group 1 (P < 0.05). The sensitivity and specificity of PF/S sialic acid ratio with cut off value of 0.7 were 76.67% and 20%, respectively, whereas taking the cut off value of 70mg/dl for pleural fluid sialic acid in malignant pleural effusions, the sensitivity was 63.33%, specificity 60%, and positive predictive value 46.34%. Conclusion: These findings indicate that determination of sialic acid levels in pleural fluid has diagnostic value as a cheap, simple, and reliable marker for malignant pleural effusion.
Lessons learned from the SYNTAX trial for multivessel and left main stem coronary artery disease

Purpose of review The SYNergy between percutaneous coronary intervention with TAXus and cardiac surgery (SYNTAX) trial is the most important trial of surgery and stents in patients with severe coronary artery disease (CAD) and reflects real clinical practice. This review describes its key findings at 3 years with particular reference to what is already known on this topic. Recent findings Taking together the SYNTAX randomized trial and its registry component, almost 79% of patients with three-vessel CAD and almost two-thirds of patients with left main stem (LMS) disease have a survival benefit and marked reduction in the need for repeat revascularization with coronary artery bypass grafting (CABG) in comparison to stents, implying that CABG is still the treatment of choice for most of these patients. This conclusion, at odds with results of previous trials of stenting and surgery but consistent with findings of large propensity-matched registries, can be explained by the fact that SYNTAX enrolled ‘real life’ patients rather than the highly select patients in previous trials. SYNTAX also shows that for patients with less severe CAD there is no difference in survival between CABG and stents but a lower incidence of repeat revascularization with CABG. Summary The results of the SYNTAX trial confirm that at 3 years CABG remains the treatment of choice for most patients with three-vessel and LMS disease and especially in those with the most severe disease. SYNTAX will have a profound effect on practice recommendations for the foreseeable future and already has had a major effect on the new European Society for Cardiology/European Association for Cardiothoracic Surgery guidelines for myocardial revascularization.
Gene expression profiling to define host response to baculoviral transduction in the brain

Recombinant baculoviral vectors efficiently transduce several types of cells in the brain. To characterize host responses to virus challenge, thus verifying the suitability of using baculovirus for the development of gene therapy strategies in the central nervous system, we used cDNA microarray technology to examine in vitro and in vivo global cellular gene expression profiles in the rat brain, cultured human astrocytes and human neuronal cells after viral transduction. We demonstrated that the transduction induced host antiviral responses as a major reaction in all three types of samples profiled. The related genes were mainly those associated with innate immunity, including several of the genes involved in Toll‐like receptor signaling pathway and cytokine‐cytokine receptor interaction. These findings should be useful in understanding the molecular basis for neural cell response to baculoviral transduction and in guiding rational therapeutic applications of baculoviral vectors in the central nervous systems.
Rapid constitutive and ligand‐activated endocytic trafficking of P2X3 receptor

P2X receptors mediate a variety of physiological actions, including smooth muscle contraction, neuro‐endocrine secretion and synaptic transmission. Among P2X receptors, the P2X3 subtype is expressed in sensory neurons of dorsal root‐ and trigeminal‐ganglia, where it performs a well‐recognized role in sensory and pain transmission. Recent evidence indicates that the strength of P2X3‐mediated responses is modulated in vivo by altering the number of receptors at the plasma membrane. In the present study, we investigate the trafficking properties of P2X3 receptor in transfected HEK293 cells and in primary cultures of dorsal root ganglion neurons, finding that P2X3 receptor undergoes rapid constitutive and cholesterol‐dependent endocytosis. We also show that endocytosis is accompanied by preferential targeting of the receptor to late endosomes/lysosomes, with subsequent degradation. Furthermore, we observe that at steady state the receptor localizes predominantly in lamp1‐positive intracellular structures, with a minor fraction present at the plasma membrane. Finally, the level of functional receptor expressed on the cell surface is rapidly up‐regulated in response to agonist stimulation, which also augments receptor endocytosis. The findings presented in this work underscore a very dynamic trafficking behavior of P2X3 receptor and disclose a possible mechanism for the rapid modulation of ATP‐mediated responses potentially relevant during physiological and pathological conditions.
Apolipoprotein‐D expression is increased during development and maturation of the human prefrontal cortex

Apolipoprotein D (apoD) is a lipid binding protein expressed in the brain where its function is largely unknown. Based on changes in lipid metabolism and deposition that occur in the human brain during postnatal development, we investigated changes in apoD expression in the prefrontal cortex in 69 normal cases ranging in age from 40 days to 49 years utilizing gene microarray, quantitative PCR and western blotting methods. In contrast to the high expression of apolipoprotein E (APOE), low‐density lipoprotein receptor‐related protein 8 (LRP8) and 3‐hydroxy‐3‐methyl‐glutaryl‐CoA reducatase (HMGCR) (genes that play a role in lipid‐related pathways in brain development) early in life, apoD expression was low in neonates and increased in expression throughout life resulting in six‐ to eight‐fold higher levels at the mRNA and protein levels in adults. Recent studies suggest that apoD has a novel antioxidant function in the brain and we found that the increased apoD expression throughout development and into adulthood was correlated with the expression of antioxidant genes superoxide dismutase 1 (SOD1) and glutathione peroxidase 3 (GPX3) as well as proteins that were modified by the lipid peroxidation end‐product 4‐hydroxynonenal. These studies reveal that apoD expression is increased throughout life in the human prefrontal cortex and that this is correlated with genetic and biochemical markers of oxidative stress.
Nerve cells culture from lumbar spinal cord on surfaces modified by plasma pyrrole polymerization

Currently, there are several techniques for modified cell culture surfaces under research to improve cell growth and adhesion. Recently, different methods have been used for surface coating, using biomolecules that enhance cell attachment and growth of nerve cells from spinal cord, such as the use of Poly-DL-Ornithine/Laminin. Plasma-polymerized pyrrole (PPy)-treated surfaces have showed improvement on surfaces biocompatibility with the cells in culture since they do not interfere with any of the biological cell functions. In the present work, we present a novel mouse nerve cell culture technique, using PPy-treated cell culture surfaces. A comparative study of cell survival using Poly-DL-Ornithine/Laminin-treated surfaces was performed. Our results of cell survival when compared with data already reported by other investigators, show that cells cultured on the PPy-modified surface increased survival up to 21 days when compared with Poly-DL-Ornithine/Laminin-coated culture, where 8 days cell survival was obtained. There were electrical and morphological differences in the nerve cells grown in the different surfaces. By comparing the peak ion currents of Poly-DL-Ornithine/Laminin-seeded cells for 8 days with cells grown for 21 days on PPy, an increase of 516% in the Na+ current and 127% in K+ currents in cells seeded on PPy were observed. Immunofluorescence techniques showed the presence of cell synapses and culture viability after 21 days. Our results then showed that PPy-modified surfaces are an alternative culture method that increases nerve cells survival from lumbar spinal cord cell culture by preserving its electrical and morphological features.
The importance of increased percentage of biventricular pacing to improve clinical outcomes in patients receiving cardiac resynchronization therapy

Purpose of review To describe the growing evidence that a maximal biventricular pacing is needed to gain the maximal benefits from cardiac resynchronization therapy (CRT). Recent findings Even small gains in the biventricular (BIV) pacing percentage are clinically important both to prevent acute heart failure and, more importantly, to improve survival. Summary Every effort should be made in all patients receiving CRT to approach 100% BIV pacing by a correct device programming, a correct pharmacologic regimen and atrioventricular nodal ablation in atrial fibrillation patients.
A metabolic switch in brain: glucose and lactate metabolism modulation by ascorbic acid

In this review, we discuss a novel function of ascorbic acid in brain energetics. It has been proposed that during glutamatergic synaptic activity neurons preferably consume lactate released from glia. The key to this energetic coupling is the metabolic activation that occurs in astrocytes by glutamate and an increase in extracellular [K+]. Neurons are cells well equipped to consume glucose because they express glucose transporters and glycolytic and tricarboxylic acid cycle enzymes. Moreover, neuronal cells express monocarboxylate transporters and lactate dehydrogenase isoenzyme 1, which is inhibited by pyruvate. As glycolysis produces an increase in pyruvate concentration and a decrease in NAD+/NADH, lactate and glucose consumption are not viable at the same time. In this context, we discuss ascorbic acid participation as a metabolic switch modulating neuronal metabolism between rest and activation periods. Ascorbic acid is highly concentrated in CNS. Glutamate stimulates ascorbic acid release from astrocytes. Ascorbic acid entry into neurons and within the cell can inhibit glucose consumption and stimulate lactate transport. For this switch to occur, an ascorbic acid flow is necessary between astrocytes and neurons, which is driven by neural activity and is part of vitamin C recycling. Here, we review the role of glucose and lactate as metabolic substrates and the modulation of neuronal metabolism by ascorbic acid.
Muscleblind‐like 2: circadian expression in the mammalian pineal gland is controlled by an adrenergic‐cAMP mechanism

Muscleblind‐like 2 (Mbnl2) is a zinc finger protein first identified in Drosophila. It appears to be essential for photoreceptor development and to be involved in RNA splicing. Here we report that Mbnl2 is strongly expressed in the rat pineal gland. The abundance of pineal Mbnl2 transcripts follows a marked circadian rhythm with peak levels approximately sevenfold higher at night than day levels. Mbnl2 protein exhibits a similar rhythm. In vitro studies indicate that the abundance of Mbnl2 transcripts and protein are controlled by an adrenergic/cAMP mechanism.
Risk Factors for Malnutrition Among Under-Five-Year olds in an Inner City Community inIbadan: A Case-Control Study

Background:Morbidity and mortality rates associated with malnutrition in the under-five-year olds are high especially among children from the low socio-economic class, yet not all children from this deprived environment develop malnutrition. Objectives: To identify risk factors associated with the development of malnutrition in the under-five-year olds in a homogeneous inner city community. Design: A community-based, case-control study. Materials and Methods: One hundred and thirty eight children (subjects and controls) aged less than five years living in the traditional area of Idikan, Ibadan, participated in the study. All households with children under five years were listed and visited over a period of three months. All children in these households were examined and those with weight-for-age less than 2 standard deviations of the NCHS median value were enrolled as subjects. A control, who was not malnourished, was selected for every subject identified; they were matched 1:1 by age and sex from the same compound or adjacent compounds. Results: The youngest malnourished child was two months old while the mean age of the subjects was 22.5 ± 14.1 months. A significantly higher proportion of subjects than controls had primary caretakers who were not their parents (16.9 percent vs 6.2 percent; p 0.05). In the case-control analysis, low paternal education (incomplete primary school education and less)(p Conclusion: Infants should be breastfed exclusively for the first six months of life, and breastfeeding should continue up to the age of two years. Immunization coverage needs to be improved in the community. These measures should help in reducing malnutrition in the community. Keywords: Malnutrition, Under-five morbidity, Low socio-economic environment, Immunization, Nigerian Journal of Paediatrics 2003. 30:7-12
Brain sterol dysregulation in sporadic AD and MCI: relationship to heme oxygenase‐1

The objective of this study was to ascertain the impact of aging and Alzheimer’s disease (AD) on brain cholesterol (CH), CH precursors, and oxysterol homeostasis. Altered CH metabolism and up‐regulation of heme oxygenase‐1 (HO‐1) are characteristic of AD‐affected neural tissues. We recently determined that HO‐1 over‐expression suppresses total CH levels by augmenting liver X receptor‐mediated CH efflux and enhances oxysterol formation in cultured astroglia. Lipids and proteins were extracted from postmortem human frontal cortex derived from subjects with sporadic AD, mild cognitive impairment (MCI), and no cognitive impairment (n = 17 per group) enrolled in the Religious Orders Study, an ongoing clinical‐pathologic study of aging and AD. ELISA was used to quantify human HO‐1 protein expression from brain tissue and gas chromatography–mass spectrometry to quantify total CH, CH precursors, and relevant oxysterols. The relationships of sterol/oxysterol levels to HO‐1 protein expression and clinical/demographic variables were determined by multivariable regression and non‐parametric statistical analyses. Decreased CH, increased oxysterol and increased CH precursors concentrations in the cortex correlated significantly with HO‐1 levels in MCI and AD, but not no cognitive impairment. Specific oxysterols correlated with disease state, increasing neuropathological burden, neuropsychological impairment, and age. A model featuring compensated and de‐compensated states of altered sterol homeostasis in MCI and AD is presented based on the current data set and our earlier in vitro work.
A protein–protein interaction of stress‐responsive myosin VI endowed to inhibit neural progenitor self‐replication with RNA binding protein, TLS, in murine hippocampus

We have shown preferential expression of both mRNA and corresponding protein for myosin VI (Myo6) in the murine hippocampus within 24 h after the extreme traumatic experience, water‐immersion restraint stress (WIRS), prior to a drastic decrease in neural progenitor proliferation in the dentate gyrus. Myosin (Myo6) protein levels were significantly increased in hippocampus within 24 h after flashback experience in mice previously exposed to WIRS. Myo6 protein was ubiquitously distributed in discrete mouse brain regions with exceptionally high expression in olfactory bulb, whereas Myo6 protein was expressed in cultured rat astroglia and neurons, in addition to Myo6 mRNA expression by cultured neural progenitors. In mouse embryonal carcinoma P19 cells endowed to proliferate and differentiate, Myo6 protein was expressed in line with astroglial marker protein expression. Transient over‐expression of Myo6 induced a significant decrease in the size of clustered aggregates as an index of self‐replication in P19 cells. Immunoprecipitation analysis revealed the interaction between Myo6 and the RNA‐binding protein, translocated in liposarcoma (TLS), while TLS was predominantly expressed by neurons in the cortex, striatum, cerebellum, and hippocampus. These results suggest that Myo6 may play a pivotal role in the mechanism underlying the suppressed adult neurogenesis after traumatic stress in association with TLS.