Miracle-dz/UniFAR-FTUs huggingface dataset:
Postthaw viability of precultured hepatocytes.

Hepatocytes are being studied for a wide variety of applications, including drug metabolism studies, gene therapy, and use in liver-assist devices for temporary liver support. The ability to cryopreserve isolated hepatocytes would permit the pooling of cells to reach the required therapeutic coordination of the cell supply with patient care regimes and the completion of safety and quality-control testing. The objective of this investigation was to develop a method of cryopreserving isolated hepatocytes that will retain high levels of function and facilitate the use of the cells in different applications. Freshly isolated hepatocytes were cultured in a spinner flask for different periods of time, up to 48 h. The cells were cryopreserved by use of a range of solution concentrations and cooling rates. For fresh, nonfrozen hepatocytes precultured for 24 h prior to being plated on collagen, the albumin secretion rate was 0.88 +/- 0.62 mg/ml/h. When the cells were precultured for 24 h, frozen in a solution containing 10% Me2SO with a cooling rate of 1 degrees C/min, thawed, plated on collagen, and cultured, the albumin secretion rate was 0.21 +/- 0.24 microg/ml/h. In contrast, freshly isolated hepatocytes cryopreserved without preculture and cultured on collagen had an albumin secretion rate of 0.07 +/- 0.08 mg/ml/h. The influences of different solution compositions and cooling rates on postthaw function of precultured hepatocytes were also determined. These results indicate that the use of a preliminary culture step prior to cryopreservation can enhance the postthaw function of hepatocytes.
Design, Analysis, and Multicriteria Optimization of an Overground Pediatric Robotic Gait Trainer

Robotic gait training has been gathering much attention and has demonstrated potential to improve walking patterns for children with cerebral palsy. This paper proposes a novel pediatric robotic overground gait trainer (Pro-GaiT) based on a nonanthropomorphic linkage design and walking frame. The robot configuration is a 2 degree-of-freedom 5 bar linkage “end effector” and considers the human as part of the kinematic chain. This permits the user to move relative to the robot, inherently handling joint misalignments (unlike exoskeletons), to simplify clinical application while still allowing various gait patterns to be accurately achieved. To successfully achieve this complex motion, the kinematic design is critically analyzed to ensure that desired motion and forces can be imparted on the user. A thorough design analysis is conducted, and a genetic algorithm is used to optimize the robot configuration in terms of workspace, singularities, and physical size of the robot. The device was experimentally validated with four healthy subjects, as well as with clinical data from three children with CP that walk with crouch gait with varying human–robot misalignment and joint spasticity. The results demonstrate that the Pro-GaiT has achieved the desired clinical specifications.
Measurement of 5‐HT1A receptor binding in depressed adults before and after antidepressant drug treatment using positron emission tomography and [11C]WAY‐100635

Objective: To assess effects of chronic antidepressant drug treatment on serotonin type‐1A receptor (5‐HT1AR) binding potential (BP) in major depressive disorder. Methods: Depressed subjects (n = 27) were imaged using PET and [11C]WAY‐100635 at baseline and following a median of 9.4 weeks of treatment with selective serotonin reuptake inhibitor or dual reuptake inhibitor antidepressant agents. Fifteen subjects had complete pre‐ and post‐treatment scan data. The 5‐HT1AR BP was derived from the tissue time‐radioactivity concentrations from regions‐of‐interest defined a priori, using a simplified reference tissue model (SRTM), and in a subset of subjects, compartmental modeling (CMOD). Results: Chronic treatment had no effect on pre‐ or post‐synaptic 5‐HT1AR BP, as confirmed by both the SRTM and CMOD analyses. These results were unaffected by treatment response status and were consistent across brain regions. Among the 22 subjects for whom the clinical response‐to‐treatment was established, the treatment nonresponders (n = 7) had higher baseline BP values in the left (P = 0.01) and right orbital cortex (P = 0.02) than the responders (n = 15). Conclusions: Chronic antidepressant drug treatment did not significantly change cerebral 5‐HT1AR binding, consistent with preclinical evidence that the alterations in serotonergic function associated with antidepressant drug administration are not accompanied by changes in 5‐HT1AR density. Higher baseline 5‐HT1AR binding was associated with poorer response to treatment. Synapse 61:523–530, 2007. © 2007 Wiley‐Liss, Inc.
A Novel Deadbeat Current Control Scheme with an Adaptive Self-Tuning Load Model for a Three-Phase PWM-VSI

In this paper, a novel deadbeat current control scheme with an adaptive self-tuning load model for a three-phase pulse-width-modulated (PWM) voltage source inverter (VSI) is proposed. First, to achieve high bandwidth current control characteristics, an improved deadbeat current controller with delay compensation is adopted. The compensation method forces the delays elements, caused by voltage calculation, PWM, and synchronous frame rotation, to be equivalently placed outside the close loop control system. Hence, their effect on the closed loop stability is eliminated and the current controller can be designed with a higher bandwidth. Second, to relax the parameters sensitivity issue of the deadbeat controller and to realize a control scheme with reduced sensors, a novel adaptive self-tuning load model is emerged in the control structure. The adaptive model is designed with low computational demand to estimate, in the real-time, the load parameters (R, L) and the back EMF voltage simultaneously. A unified solution to the present nonlinear estimation problem is presented by adopting a parallel observers structure. Furthermore, the adaptive model has the necessary phase advance of the estimated quantities; which compensates for the total system's delay. Comparative evaluation results are presented to demonstrate the validity and effectiveness of the proposed control scheme.
Enhancement of methane yield from wheat straw, miscanthus and willow using aqueous ammonia soaking

The increasing demand for methane production cannot be satisfied by the use of anaerobic digestion only from waste/wastewater treatment. Perennial energy crops, such as miscanthus and willow, as well as agricultural residues can be considered as options for increasing the methane production through biomass digestion, due to their high organic content and biomass yield. These materials present a great potential, which is only limited by the rigid lignocellulosic structure. In this case, it is possible to apply a pretreatment step in order to achieve increased biogas production. In the present study, aqueous ammonia soaking (AAS) has been investigated as a method to disrupt the lignocellulosic structure and increase the methane yield of wheat straw, miscanthus and willow. Among the three biomasses tested, wheat straw and miscanthus were the most promising in terms of methane production, yielding around 200 and 230 ml of methane per gram of total solids. In all three cases, AAS resulted to an increase in methane yield of 37–41%, 25–27% and 94–162% for wheat straw, miscanthus and willow, respectively. A comparison of the methane yields after 20 and 50 days of anaerobic digestion revealed that AAS affected positively the methane production rate as well. AAS also resulted to a low solubilization of sugars, with a 15.4% and 8.9% increase in soluble xylose concentration in miscanthus and willow, respectively, and a 5% solubilization of glucose in AAS-pretreated miscanthus.
Risk factors of mortality in perforated peptic ulcer.

OBJECTIVE To assess the risk factors that influence mortality from perforated peptic ulcer. DESIGN Retrospective study. SETTING General hospital, Taiwan. SUBJECTS 179 patients who had their perforated peptic ulcers operated on and who had minimum follow-up of one year. MAIN OUTCOME MEASURES Mortality. RESULTS The overall mortality was 15% (26/179). Of the 26 patients who died, the cause of death was uncontrolled systemic infection in 21 (81%), hypovolaemic shock in 2, and fatal arrhythmia and heart failure in 1 each. 15 of the patients who died of sepsis did not have fulminant abdominal sepsis. Most deaths occurred early after operation, (range 1-96 days). Old age, preoperative shock, and type of operation seemed to be related to these deaths on univariate analysis, but multivariate analysis showed that coexisting medical illness, delayed treatment, and low albumin concentration were independent risk factors for mortality. CONCLUSIONS To improve the result of treatment of perforated peptic ulcer, the diagnosis and treatment should not be delayed, the associated medical illnesses should be treated, and nutritional support should be given.
High frequencies of circulating IFN‐γ‐secreting CD8 cytotoxic T cells specific for a novel MHC class I‐restricted Mycobacterium tuberculosis epitope in M. tuberculosis‐infected subjects without disease

MHC class I‐restricted CD8 cytotoxic T lymphocytes (CTL) are essential for protective immunity to Mycobacterium tuberculosis in animal models but their role in humans remains unclear. We therefore studied subjects who had successfully contained M. tuberculosis infection in vivo, i. e. exposed healthy household contacts and individuals with inactive self‐healed pulmonary tuberculosis. Using the ELISPOT assay for IFN‐γ, we screened peptides from ESAT‐6, a secreted antigen that is highly specific for M. tuberculosis. We identified a novel nonamer epitope: unstimulated peripheral blood‐derived CD8 T cells displayed peptide‐specific IFN‐γ release ex vivo while CD8 T cell lines and clones exhibited HLA‐A68.02‐restricted cytolytic activity and recognized endogenously processed antigen. The frequency of CD8 CTL specific for this single M. tuberculosis epitope, 1 / 2500 peripheral blood lymphocytes, was equivalent to the combined frequency of all IFN‐γ‐secreting purified protein derivative‐reactive T cells ex vivo. This highly focused CTL response was maintained in an asymptomatic contact over 2 years and is the most potent antigen‐specific anti‐mycobacterial CD8 CTL response hitherto described. Thus, human M. tuberculosis‐specific CD8 CTL are not necessarily associated with active disease per se. Rather, our results are consistent with a protective role for these ESAT‐6‐specific CD8 T cells in the long‐term control of M. tuberculosis in vivo in humans.
Contrasting effects of cerebrospinal fluid from motor neuron disease patients on the survival of primary motor neurons cultured with or without glia

Abstract Motor neuronal (MN) degeneration in motor neuron disease (MND) often starts focally before spreading to neighbouring MN populations, suggesting soluble factors may contribute to disease propagation. Whether cerebrospinal fluid (CSF) from MND patients contains such factors has been difficult to prove. We aimed to determine the effect of glia on the response of MNs to CSF from MND patients. Primary rat spinal MNs grown in mono-culture or cocultured with glia were exposed to CSF from patients (MND-CSF) or controls (Con-CSF) and survival measured by cell counting. In mono-culture both MND-CSF and Con-CSF reduced MN survival with MND-CSF reducing MN survival by less than Con-CSF. In coculture MN survival was unchanged by exposure to MND-CSF while exposure to Con-CSF improved MN survival. In separate experiments, murine MNs grown in mono-culture and stressed by growth factor withdrawal were partially rescued by the application of monocyte chemoattractant protein-1 (MCP-1), a trophic factor previously found to be elevated in MND-CSF. Our results suggest that MND-CSF may contain factors harmful to MNs as well as factors protective of MNs, the interplay of which is altered by the presence of glial cells. These preliminary results further emphasize the importance of MN environment to MN health.
Management of Bipolar II Disorder

Bipolar II disorder (BP II) disorder was recognized as a distinct subtype in the DSM-IV classification. DSM-IV criteria for BP II require the presence or history of one or more major depressive episode, plus at least one hypomanic episode, which, by definition, must last for at least 4 days. Various studies found distinct patterns of symptoms and familial inheritance for BP II disorder. BP II is commonly underdiagnosed or misdiagnosed. Making an early and accurate diagnosis of BP II is utmost importance in the management of BP II disorder. The clinician should have this diagnosis in mind when he is facing a patient presenting with mood problems, particularly unipolar depression. Quetiapine and lamotrigine are the only agents with demonstrated efficacy in double-blind RCT. Although the evidence for the use of lithium in long-term therapy is largely based on observational studies, the many years of close follow-up, comparatively larger subject numbers, and ‘harder’ clinically meaningful with bipolar disorder outcomes measures, enhance our confidence in its role in treating BP II. With respect to short-term treatment, there is some limited support for the use of risperidone and olanzepine in hypomania and for fluoxetine, venlafaxine and valproate in treating depression. The current clinical debate over whether one should use antidepressants as monotherapy or in combination with a mood stabilizer when treating BP II depression is not yet settled. There is a need for large, well-designed RCTs to cast more definitive light on how best to manage patients with BP II disorder.
Performance in a Collaborative Search Task: The Role of Feedback and Alignment

When people communicate, they coordinate a wide range of linguistic and non-linguistic behaviors. This process of coordination is called alignment, and it is assumed to be fundamental to successful communication. In this paper, we question this assumption and investigate whether disalignment is a more successful strategy in some cases. More specifically, we hypothesize that alignment correlates with task success only when communication is interactive. We present results from a spot-the-difference task in which dyads of interlocutors have to decide whether they are viewing the same scene or not. Interactivity was manipulated in three conditions by increasing the amount of information shared between interlocutors (no exchange of feedback, minimal feedback, full dialogue). We use recurrence quantification analysis to measure the alignment between the scan-patterns of the interlocutors. We found that interlocutors who could not exchange feedback aligned their gaze more, and that increased gaze alignment correlated with decreased task success in this case. When feedback was possible, in contrast, interlocutors utilized it to better organize their joint search strategy by diversifying visual attention. This is evidenced by reduced overall alignment in the minimal feedback and full dialogue conditions. However, only the dyads engaged in a full dialogue increased their gaze alignment over time to achieve successful performances. These results suggest that alignment per se does not imply communicative success, as most models of dialogue assume. Rather, the effect of alignment depends on the type of alignment, on the goals of the task, and on the presence of feedback.
Cancer translocations in human cells induced by zinc finger and TALE nucleases

Chromosomal translocations are signatures of numerous cancers and lead to expression of fusion genes that act as oncogenes. The wealth of genomic aberrations found in cancer, however, makes it challenging to assign a specific phenotypic change to a specific aberration. In this study, we set out to use genome editing with zinc finger (ZFN) and transcription activator-like effector (TALEN) nucleases to engineer, de novo, translocation-associated oncogenes at cognate endogenous loci in human cells. Using ZFNs and TALENs designed to cut precisely at relevant translocation breakpoints, we induced cancer-relevant t(11;22)(q24;q12) and t(2;5)(p23;q35) translocations found in Ewing sarcoma and anaplastic large cell lymphoma (ALCL), respectively. We recovered both translocations with high efficiency, resulting in the expression of the EWSR1–FLI1 and NPM1–ALK fusions. Breakpoint junctions recovered after ZFN cleavage in human embryonic stem (ES) cell–derived mesenchymal precursor cells fully recapitulated the genomic characteristics found in tumor cells from Ewing sarcoma patients. This approach with tailored nucleases demonstrates that expression of fusion genes found in cancer cells can be induced from the native promoter, allowing interrogation of both the underlying mechanisms and oncogenic consequences of tumor-related translocations in human cells. With an analogous strategy, the ALCL translocation was reverted in a patient cell line to restore the integrity of the two participating chromosomes, further expanding the repertoire of genomic rearrangements that can be engineered by tailored nucleases.
Multilevel Power-Imbalance Allocation Control for Secondary Frequency Control of Power Systems

A consensus-control-based multilevel control law named Multilevel Power-Imbalance Allocation Control (MLPIAC) is presented for a large-scale power system partitioned into two or more groups. Centralized control is implemented in each group while distributed control is implemented at the coordination level of the groups. Besides restoring nominal frequency with a minimal control cost, MLPIAC can improve the transient performance of the system through an accelerated convergence of the control inputs without oscillations. At the coordination level of the control groups, because the number of the groups is smaller than that of nodes, MLPIAC is more effective to obtain the minimized control cost than the purely distributed control law. At the level of the control in each group, because the number of nodes is much smaller than the total number of nodes in the whole network, the overheads in the communications and the computations are reduced compared to the pure centralized control. The asymptotic stability of MLPIAC is proven using the Lyapunov method and the performance is evaluated through simulations.
Immediate results of bifurcational stenting assessed with optical coherence tomography

Background: A tubular stent may adapt with difficulty to coronary bifurcation lesions (CBLs). Methods: Time domain or frequency domain (FD) optical coherence tomography (OCT) was performed to assess strut apposition immediately after stent implantation across four segments inside the bifurcation, in a consecutive series of patients. OCT pullbacks were performed in the main vessel (MV). Results: A total of 13,142 struts in 45 CBL in 41 patients were assessed. Strut malapposition was significantly more frequent in the half bifurcation facing the side‐branch (SB) ostium (42.9%) than in the proximal segment of the bifurcation 11.8%, half bifurcation opposite the SB 6.7%, or the distal segment 5.7% (all P < 0.0001). Lesions (n = 15) treated with stenting of both MV and SB had a total higher rate of malapposition than those (n = 30) treated with stenting of the MV only (17.6% vs. 9.5%; P = 0.0014). In latter group, lesions treated with FD‐OCT‐guided stent implantation (n = 13) presented a lower rate of malapposition than those treated with conventional angiographic‐guided stent implantation (n = 17) (7.1% vs. 17.5%; P = 0.005). Conclusions: In CBL, strut malapposition is particularly high at the SB ostium. However, a strategy of stenting MV only with adjunctive FD‐OCT guidance is associated with lower rates of malapposition. © 2012 Wiley Periodicals, Inc.
Role of Intrachain Disulfides in the Activities of the CdtA and CdtC Subunits of the Cytolethal Distending Toxin of Actinobacillus actinomycetemcomitans

ABSTRACT The cytolethal distending toxin (Cdt) of Actinobacillus actinomycetemcomitans is an atypical A-B-type toxin consisting of a heterotrimer composed of the cdtA, cdtB, and cdtC gene products. The CdtA and CdtC subunits form two heterogeneous ricin-like lectin domains which bind the holotoxin to the target cell. Point mutations were used to study CdtC structure and function. One (mutC216F97C) of eight single-amino-acid replacement mutants identified yielded a gene product that failed to form biologically active holotoxin. Based on the possibility that the F97C mutation destabilized a predicted disulfide, targeted mutagenesis was used to examine the contribution of each of four cysteine residues, in two predicted disulfides (C96/C107 and C135/C149), to CdtC activities. Cysteine replacement mutations in two predicted disulfides (C136/C149 and C178/C197) in CdtA were also characterized. Flow cytometry and CHO cell proliferation assays showed that changing either C96 or C149 in CdtC to alanine abolished the biological activity of holotoxin complexes. However, replacing C107 or C135 in CdtC and any of the four cysteines in CdtA with alanine or serine resulted in only partial or no loss of holotoxin activity. Changes in the biological activities of the mutant holotoxins correlated with altered subunit binding. In contrast to elimination of the B chain of ricin, the elimination of intrachain disulfides in CdtC and CdtA by genetic replacement of cysteines destabilizes these subunit proteins but not to the extent that cytotoxicity is lost. Reduction of the wild-type holotoxin did not affect cytotoxicity, and the reduced form of wild-type CdtA exhibited a statistically significant increase in binding to ligand. A diminished role for intrachain disulfides in stabilizing CdtA and CdtC may have clinical relevance for the A. actinomycetemcomitans Cdt. The cdt gene products secreted by this pathogen assemble and bind to target cells in periodontally involved sites, which are decidedly reduced environments in the human oral cavity.
The ADP‐dependent sugar kinase family: Kinetic and evolutionary aspects

Some archaea of the Euryarchaeota present a unique version of the Embden–Meyerhof pathway where glucose and fructose‐6‐phosphate are phoshporylated using ADP instead of ATP as the phosphoryl donor. These are the only ADP‐dependent kinases known to date. Although initially they were believed to represent a new protein family, they can be classified as members of the ribokinase superfamily, which also include several ATP‐dependent kinases. As they were first identified in members of the thermococcales it was proposed that the presence of these ADP‐dependent kinases is an adaptation to high temperatures. Later, homologs of these enzymes were identified in the genomes of mesophilic and thermophilic methanogenic archaea and even in the genomes of higher eukaryotes, suggesting that the presence of these proteins is not related to the hyperthermophilic life. The ADP‐dependent kinases are very restrictive to their ligands being unable to use triphosphorylated nucleotides such as ATP. However, it has been shown that they can bind ATP by competition kinetic experiments. The hyperthermophilic methanogenic archaeon Methanocaldococcus jannaschii has a homolog of these genes, which can phosphorylate glucose and fructose‐6‐phosphate. For this reason, it was proposed as an ancestral form for the family. However, recent studies have shown that the ancestral activity in the group is glucokinase, and a combination of gene duplication and lateral gene transfer could have originated the two paralogs in this member of the Euryarchaeota. Interestingly, based on structural comparisons made within the superfamily it has been suggested that the ADP‐dependent kinases are the newest in the group. In several members of the superfamily, the presence of divalent metal cations has been shown to be crucial for catalysis, so its role in the ADP‐dependent family was investigated through molecular dynamics. The simulation shows that, in fact, the metal coordinates the catalytic ensemble and interacts with crucial residues for catalysis. © 2009 IUBMB IUBMB Life, 61(7): 753–761, 2009