Chemosensitization by a non-apoptogenic heat shock protein 70-binding apoptosis-inducing factor mutant

Heat shock protein 70 (HSP70) inhibits apoptosis and thereby increases the survival of cells exposed to a wide range of lethal stimuli. HSP70 has also been shown to increase the tumorigenicity of cancer cells in rodent models. The protective function of this chaperone involves interaction and neutralization of the caspase activator apoptotic protease activation factor-1 and the mitochondrial flavoprotein apoptosis-inducing factor (AIF). In this work, we determined by deletional mutagenesis that a domain of AIF comprised between amino acids 150 and 228 is engaged in a molecular interaction with the substrate-binding domain of HSP70. Computer calculations favored this conclusion. On the basis of this information, we constructed an AIF-derived protein, which is cytosolic, noncytotoxic, yet maintains its capacity to interact with HSP70. This protein, designated ADD70, sensitized different human cancer cells to apoptosis induced by a variety of death stimuli by its capacity to interact with HSP70 and therefore to sequester HSP70. Thus, its chemosensitizing effect was lost in cells in which inducible HSP70 genes had been deleted. These data delineate a novel strategy for the selective neutralization of HSP70.     

Snoring and Sleep Apnea in Obese Adolescents: Effect of Long-term Weight Loss-Rehabilitation

Objective: To test the effect of a long-term weight loss rehabilitation program in extremely obese adolescents on breathing parameters during sleep. Methods: Thirty-eight extremely obese [mean body mass index (BMI) 45.3 ± 7.9kg/m²] adolescents participated during a three- to nine-month period in an inpatient weight loss rehabilitation in a specialized long-term rehabilitation center. Breathing parameters were registered via a seven-channel portable screening device. Body weight and arterial blood pressure were measured before and after the long-term treatment. Results: Mean BMI decreased from 45.3 to 35.8 (p < 0.001), mean diastolic blood pressure decreased from 89 mmHg to 81 mmHg (p = 0,002). Nine patients had a RDI of 5 and 30 patients a RDI of <5; the mean RDI decreased from 4.08 to 3.27 (n.s.). Within the group, the RDI was 5 and the mean RDI decreased from 10.3/h to 5.2/h (p = 0.02). The mean SaO₂ increased from 93.65 to 95.35% (p = 0.003), lowest SaO₂ increased from 72.14 to 73.19% (n.s.) and snoring frequency decreased from 37.56% of total sleep time (TST) to 32.86% of TST (n.s.). Conclusion: A long-term inpatient weight loss program has a positive effect on breathing parameters during sleep in extremely obese adolescents. However, the effect on apneic events and snoring is relatively minor compared to the effect on arterial oxygen saturation. The role of obesity in the origin of respiratory events and snoring in adolescents might be overestimated.     

Energy Metabolism of Rat Skeletal Muscle Modulated by the Rate of Perfusion Flow

In order to advance our understanding of the phenomenon of flow-induced increases in the metabolism of the relaxed muscle, the metabolic rate of the isolated rat gracilis muscle was investigated at 28 degrees C in vitro. The muscle was perfused with cell-free Krebs-Henseleit bicarbonate buffer containing 5% bovine serum albumin and 5 mM glucose, saturated with a gas mixture of 95% O2 and 5% CO2 and simultaneously superfused with a medium saturated with with a low O2 gas mixture (1% O2, 5% CO2 and 94% N2). Two different perfusion flow rates (0.054 and 0.100 ml min-1) have been used. Their influence on oxygen consumption and lactate production has been measured. After a 100 min perfusion period, the muscle was freeze-clamped and analysed for ATP, phosphocreatine, creatine, lactate, pyruvate, inorganic phosphate and glycogen content. The energy state of the cell and the proportions of glycolytic and mitochondrial fluxes of ATP synthesis were evaluated. During perfusion at the low flow rate of 0.054 ml min-1, the oxygen uptake was 45 +/- 9 nmol min-1 (g wet wt)-1, accompanied by a dominance of anaerobic glycolytic synthesis of ATP over mitochondrial ATP synthesis, even though the total delivery of oxygen to muscle was three times higher than oxygen consumption. Increasing the perfusion flow rate to 0.100 ml min-1 increased the oxygen uptake to 120 +/- 6 nmol min-1 (g wet wt)-1, thus leading to a prevalence of mitochondrial ATP synthesis over glycolytic ATP synthesis. The inner stores of glycogen served as the main substrate of energy metabolism and the role of exogenous substrates in the flow-stimulated increase of oxygen uptake was negligible. The increase in perfusion rate also enhanced the energy state of the muscle fibres, which was expressed either as the creatine charge or as the value of the change of Gibbs free energy of ATP hydrolysis. Data indicate that the change of perfusion flow rate per se, apart from oxygen and exogenous substrate supply, elicits changes in the regulation of energy metabolism within non-contracting skeletal muscle under open microcirculation.     

Assessing an Infant Feeding Web Site as a Nutrition Education Tool for Child Care Providers

ObjectiveDetermine child care providers' infant feeding knowledge, attitude and behavior changes after viewing the infant feeding Web site and determine the effectiveness of the Web site and bilingual educational materials.DesignIntervention and control groups completed an on-line pretest survey, viewed a Web site for 3 months, and completed an on-line posttest survey; follow-up data were also assessed.SettingColorado child care centers.ParticipantsThirty-eight child care providers.InterventionSocial learning theory-based website was evaluated by child care providers in the treatment group and providers in the control group viewed a comparable website.Main Outcome MeasuresKnowledge, attitude, and behavior changes on feeding infants breast milk, formula, and solid food; desired changes to Web site.AnalysisIndependent samples t tests, chi-square, and repeated measures.ResultsChanges in attitudes and behaviors from pre- to posttest occurred primarily in the intervention group (P < .05). At follow-up, no significant differences were found among the 3 time periods. Providers desired no changes to Web site or materials.Conclusions and ImplicationsChild care providers appeared to have adequate knowledge on feeding infants formula and breast milk, but not on hunger cues. Providers would continue to use this Web site in the future. Further research should determine if changes in knowledge, attitudes, and behaviors are sustained over time and how infant feeding cues are read and determined in child care centers.     

Multimodal Imaging Reveals Improvement of Blood Supply to an Artificial Cell Transplant Site Induced by Bioluminescent Mesenchymal Stem Cells

Purpose

An artificial site for cell or pancreatic islet transplantation can be created using a polymeric scaffold, even though it suffers subcutaneously from improper vascularisation. A sufficient blood supply is crucial for graft survival and function and can be enhanced by transplantation of mesenchymal stem cells (MSCs). The purpose of this study was to assess the effect of syngeneic MSCs on neoangiogenesis and cell engraftment in an artificial site by multimodal imaging.

Procedures

MSCs expressing a gene for luciferase were injected into the artificial subcutaneous site 7 days after scaffold implantation. MRI experiments (anatomical and dynamic contrast-enhanced images) were performed on a 4.7-T scanner using gradient echo sequences. Bioluminescent images were acquired on an IVIS Lumina optical imager. Longitudinal examination was performed for 2 months, and one animal was monitored for 16 months.

Results

We confirmed the long-term presence (lasting more than 16 months) of viable donor cells inside the scaffolds using bioluminescence imaging with an optical signal peak appearing on day 3 after MSC implantation. When compared to controls, the tissue perfusion and vessel permeability in the scaffolds were significantly improved at the site with MSCs with a maximal peak on day 9 after MSC transplantation.

Conclusions

Our data suggest that the maximal signal obtained by bioluminescence and magnetic resonance imaging from an artificially created site between 3 and 9 days after MSC transplantation can predict the optimal time range for subsequent cellular or tissue transplantation, including pancreatic islets.

     

Crossed cerebellar diaschisis in patients with acute middle cerebral artery infarction: Occurrence and perfusion characteristics

We aimed to investigate the overall prevalence and possible factors influencing the occurrence of crossed cerebellar diaschisis after acute middle cerebral artery infarction using whole-brain CT perfusion. A total of 156 patients with unilateral hypoperfusion of the middle cerebral artery territory formed the study cohort; 352 patients without hypoperfusion served as controls. We performed blinded reading of different perfusion maps for the presence of crossed cerebellar diaschisis and determined the relative supratentorial and cerebellar perfusion reduction. Moreover, imaging patterns (location and volume of hypoperfusion) and clinical factors (age, sex, time from symptom onset) resulting in crossed cerebellar diaschisis were analysed. Crossed cerebellar diaschisis was detected in 35.3% of the patients with middle cerebral artery infarction. Crossed cerebellar diaschisis was significantly associated with hypoperfusion involving the left hemisphere, the frontal lobe and the thalamus. The degree of the relative supratentorial perfusion reduction was significantly more pronounced in crossed cerebellar diaschisis-positive patients but did not correlate with the relative cerebellar perfusion reduction. Our data suggest that (i) crossed cerebellar diaschisis is a common feature after middle cerebral artery infarction which can robustly be detected using whole-brain CT perfusion, (ii) its occurrence is influenced by location and degree of the supratentorial perfusion reduction rather than infarct volume (iii) other clinical factors (age, sex and time from symptom onset) did not affect the occurrence of crossed cerebellar diaschisis.     

Unique white matter microstructural patterns in ADHD presentations—a diffusion tensor imaging study

Attention‐deficit/hyperactivity disorder predominantly inattentive (ADHD‐PI) and combined (ADHD‐C) presentations are likely distinct disorders that differ neuroanatomically, neurochemically, and neuropsychologically. However, to date, little is known about specific white matter (WM) regions differentiating ADHD presentations. This study examined differences in WM microstructure using diffusion tensor imaging (DTI) data from 20 ADHD‐PI, 18 ADHD‐C, and 27 typically developed children. Voxel‐wise analysis of DTI measurements in major fiber bundles was carried out using tract‐based spatial statistics (TBSS). Clusters showing diffusivity abnormalities were used as regions of interest for regression analysis between fractional anisotropy (FA) and neuropsychological outcomes. Compared to neurotypicals, ADHD‐PI children showed higher FA in the anterior thalamic radiations (ATR), bilateral inferior longitudinal fasciculus (ILF), and in the left corticospinal tract (CST). In contrast, the ADHD‐C group exhibited higher FA in the bilateral cingulum bundle (CB). In the ADHD‐PI group, differences in FA in the left ILF and ATR were accompanied by axial diffusivity (AD) abnormalities. In addition, the ADHD‐PI group exhibited atypical mean diffusivity in the forceps minor (FMi) and left ATR and AD differences in right CB compared to healthy subjects. Direct comparison between ADHD presentations demonstrated radial diffusivity differences in FMi. WM clusters with FA irregularities in ADHD were associated with neurobehavioral performance across groups. In conclusion, differences in WM microstructure in ADHD presentations strengthen the theory that ADHD‐PI and ADHD‐C are two distinct disorders. Regions with WM irregularity seen in both ADHD presentations might serve as predictors of executive and behavioral functioning across groups. Hum Brain Mapp 37:3323–3336, 2016. © 2016 Wiley Periodicals, Inc.     

Predictors of bone fractures in a single-centre cohort of hemodialysis patients: a 2-year follow-up study

International Urology and Nephrology - Bone involvement represents one of the complications of end-stage chronic kidney disease, with fractures being its major risk. The aim of our study was to...     

Long-lasting cognitive injury in rats with apparent full gross neurological recovery after short-term cardiac arrest

OBJECTIVE: The long-term behavioral effects of mild global ischemia have not been well described. We used short (5 min) asphyxic-cardiac arrest that resulted in no apparent gross neurological deficits to study the long-term effects of mild hypoxic ischemia on the neurobehavioral status of rats. METHODS: Fifteen adult, male Wistar rats were studied. One group was given asphyxic-cardiac arrest (CA) for 5 min (n=10) and the other group had Sham procedure (n=5). Neurobehavioral testing was performed before and 2 weeks after CA. The neurobehavioral evaluations were: neurological deficit score (NDS), Y Maze, open field, pre-pulse inhibition (PPI) of acoustic startle reflex (ASR), wire hanging, and inclined screen. RESULTS: At 24h post-CA, all of the rats regained normal neurological function as measured by NDS, an integral score for consciousness, brainstem reflexes, sensorimotor function and simple behavioral reflex tests. However, 1 week after CA, the rats exhibited significant activity reductions in the open field and in spontaneous alternation in the Y maze. The CA rats also showed a significant decrease in startle reaction amplitude and startle inhibition in the PPI tests. Two weeks after CA, the changes in motor activity and deficits in PPI remained significant, but the spontaneous alternation recovered. The muscle strength test of wire hanging and inclined screen tests did not exhibit significant change. CONCLUSION: We present a rodent model of mild CA that, despite apparent full recovery of global neurological function at 24h post-resuscitation, exhibited long-term cognitive injury lasting for at least 2 weeks after CA. This model may help understand better the injury associated with CA and develop management strategies for mild brain injury.