Miracle-dz/UniFAR-FTUs huggingface dataset:
Activation of Nuclear Factor Kappa B (NF- κ B) Assayed by Laser Scanning Cytometry (LSC)

Nuclear factor kappa B (NF- κ B)/rel is the family of ubiquitous transcriptional activators involved in regulation of diverse immune and inflammatory responses. It also plays a role in control of cell growth and apoptosis. In its inactive form NF- κ B remains in the cytoplasm sequestered through interaction with I κ B protein. Rapid translocation of NF- κ B from cytoplasm to nucleus that occurs in response to extracellular signals is considered to be a hallmark feature of its activation. The translocation of NF- κ B in HL-60, U-937 and Jurkat leukemic cells as well as in human fibroblasts induced by tumor necrosis factor α (TNF- α ) or phorbol myristate acetate (PMA) was presently measured by laser scanning cytometry (LSC). NF- κ B was detected immunocytochemically with FITC-tagged antibody and its presence in the nucleus vis-a-vis cytoplasm was monitored by measuring the green fluorescence integrated over the nucleus, which was counter-stained with propidium iodide (PI), and over the cytoplasm, respectively. Activation of NF- κ B led to a rapid increase in NF- κ B-associated fluorescence measured over the nucleus (F N ) concomitant with a decrease in fluorescence over the cytoplasm (F C ), which was reflected by an increase in F N /F C ratio. This rapid assay of NF- κ B activation can be combined with morphological identification of the activated cells or with their immunophenotype. Bivariate analysis of NF- κ B expression versus cellular DNA content makes it possible to correlate its activation with the cell cycle position. The described method has a potential to be used as a functional assay to monitor intracellular translocation of other transcriptional activators such as p53 tumor suppressor protein or signal transduction molecules. κ B molecules from cytoplasm to nucleus or vice versa. Experiments are underway in our laboratory to monitor translocation of the tumor suppressor p53 protein to the nucleus in response to damage by modalities.
Extensively and pre-extensively drug resistant tuberculosis in clinical isolates of multi-drug resistant tuberculosis using classical second line drugs (levofloxacin and amikacin).

OBJECTIVE To find out the frequency of Extensively Drug Resistant (XDR) and pre-XDR tuberculosis in clinical isolates of Multi-Drug Resistant (MDR) Tuberculosis (TB) by determining the susceptibilities against Levofloxacin and Amikacin (classical second line antituberculosis drugs). STUDY DESIGN Adescriptive cross-sectional study. PLACE AND DURATION OF STUDY Microbiology Department, Armed Forces Institute of Pathology (AFIP), Rawalpindi, from September 2011 to August 2013. METHODOLOGY Amikacin (AK) and Levofloxacin (LEVO) were obtained in chemically pure form from Sigma (Taufkirchen, Germany). The breakpoint concentration used for AK was 1.0 µg/ml and for LEVO 2.0 µg/ml. Mycobacterial Growth Indicator Tube (MGIT) 960 system was used to carry out drug susceptibility testing as per recommended protocol. RESULTS A total of 3 MDR-TB isolates (3%) turned out to be XDR-TB based upon simultaneous resistance to injectable second line antituberculosis drug AK and one of the fluoro-quinolones (LEVO). A total of 24 MDR-TB isolates (24%) were found to be pre-XDR based upon resistance to LEVO alone. Treatment status record of patients with XDR and pre-XDRTB isolates revealed that majority of patients had received fluoroquinolones (FQs) during the course of treatment. CONCLUSION XDR-TB has started to emerge in MDR-TB isolates in our set up. The worrying sign is the high frequency of pre-XDR tuberculosis. Urgent steps need to be taken to stem the tide of pre-XDR-TB in our population. It is thus recommended to develop facilities to carry out drug susceptibility testing to monitor the status of pre-XDR and XDR-TB in our population.
Use of a Combination Biomarker Algorithm To Identify Medical Intensive Care Unit Patients with Suspected Sepsis at Very Low Likelihood of Bacterial Infection

ABSTRACT Sepsis remains a diagnostic challenge in the intensive care unit (ICU), and the use of biomarkers may help in differentiating bacterial sepsis from other causes of systemic inflammatory syndrome (SIRS). The goal of this study was to assess test characteristics of a number of biomarkers for identifying ICU patients with a very low likelihood of bacterial sepsis. A prospective cohort study was conducted in a medical ICU of a university hospital. Immunocompetent patients with presumed bacterial sepsis were consecutively enrolled from January 2012 to May 2013. Concentrations of nine biomarkers (α-2 macroglobulin, C-reactive protein [CRP], ferritin, fibrinogen, haptoglobin, procalcitonin [PCT], serum amyloid A, serum amyloid P, and tissue plasminogen activator) were determined at baseline and at 24 h, 48 h, and 72 h after enrollment. Performance characteristics were calculated for various combinations of biomarkers for discrimination of bacterial sepsis from other causes of SIRS. Seventy patients were included during the study period; 31 (44%) had bacterial sepsis, and 39 (56%) had other causes of SIRS. PCT and CRP values were significantly higher at all measured time points in patients with bacterial sepsis. A number of combinations of PCT and CRP, using various cutoff values and measurement time points, demonstrated high negative predictive values (81.1% to 85.7%) and specificities (63.2% to 79.5%) for diagnosing bacterial sepsis. Combinations of PCT and CRP demonstrated a high ability to discriminate bacterial sepsis from other causes of SIRS in medical ICU patients. Future studies should focus on the use of these algorithms to improve antibiotic use in the ICU setting.
Pressure effects on the binding of vanadate to the sarcoplasmic reticulum calcium-transport enzyme.

The effect which hydrostatic pressure exerts on the binding of vanadate to the calcium-transport enzyme was determined. The recent unavailability of radioactive vanadate prevented direct measurements of vanadate binding. The vanadate-free enzyme fraction was instead monitored by phosphorylating it with ATP according to Medda and Hasselbach [Medda, P. & Hasselbach, W. (1983) Eur. J. Biochem. 137, 7-14]. Vanadate binding is reduced with rising pressure at first markedly and subsequently, above 30 MPa, relatively little. The biphasic pressure-binding relationship was analysed by applying a biexponential fitting procedure to the experimental data. The biphasicity of the pressure-binding relationship indicates that the description of vanadate binding requires at least a two-step reaction sequence. The volume increments which predominate at lower pressure values, range from 200-400 ml.mol-1 depending on the composition of the reaction medium containing 5 microM and 20 microM vanadate and no or 15% (by vol.) Me2SO. The binding volumes deduced for the higher pressure range amount to 20-40 ml.mol-1. Vanadate binding is reduced in the presence of 30 microM calcium, and simultaneously both binding volumes are diminished by 100 ml.mol-1 and 20 ml.mol-1 for the low and high pressure values, respectively, as one can expect for mutual interactions between the two ligands of the transport enzyme.
E. coli BW535, a triple mutant for the DNA repair genes xth, nth, and nfo, chronically induces the SOS response

A strong chronic induction of the SOS response system occurs in E. coli BW535, a strain defective in nth, nfo and xth genes, and hence severely deficient in the repair of abasic sites in DNA. This was shown here by visualization of filamentous growth of the BW535 strain and by measuring the level of β‐galactosidase expressed in BW535/pSK1002 in comparison to the AB1157/pSK1002 strain. The plasmid pSK1002 bears an umuC::lacZ fusion in which lacZ is under the control of the umuC promoter and regulated under the SOS regulon. Increases in the expression of β‐galactosidase occur in BW535 without any exogenous SOS inducer. Chronic induction of the SOS response was observed previously in E. coli strains bearing mutations in certain genes that have mutator activity and BW535 is a moderate mutator strain. However, not all mutators show this property, since chronic induction of SOS was not observed in mutT or mutY mutators. MutT and MutY proteins, when active, protect bacteria from mutations induced by 8‐oxoG lesions in DNA. This suggests that accumulation of abasic sites, but not 8‐oxoG residues in DNA, induce the SOS response. Environ. Mol. Mutagen. 41:237–242, 2003. © 2003 Wiley‐Liss, Inc.
MPK9 and MPK12 function in SA-induced stomatal closure in Arabidopsis thaliana

Salicylic acid (SA) induces stomatal closure sharing several components with abscisic acid (ABA) and methyl jasmonate (MeJA) signaling. We have previously shown that two guard cell-preferential mitogen-activated protein kinases (MAPKs), MPK9 and MPK12, positively regulate ABA signaling and MeJA signaling in Arabidopsis thaliana. In this study, we examined whether these two MAPKs are involved in SA-induced stomatal closure using genetic mutants and a pharmacological, MAPKK inhibitor. Salicylic acid induced stomatal closure in mpk9 and mpk12 single mutants but not in mpk9 mpk12 double mutants. The MAPKK inhibitor PD98059 inhibited SA-induced stomatal closure in wild-type plants. Salicylic acid induced extracellular reactive oxygen species (ROS) production, intracellular ROS accumulation, and cytosolic alkalization in the mpk9, mpk12, and mpk9 mpk12 mutants. Moreover, SA-activated S-type anion channels in guard cells of wild-type plants but not in guard cells of mpk9 mpk12 double mutants. These results imply that MPK9 and MPK12 are positive regulators of SA signaling in Arabidopsis guard cells.
Follow-up of breast lesions diagnosed as benign with stereotactic core-needle biopsy: frequency of mammographic change and false-negative rate.

PURPOSE To determine how often lesions diagnosed as benign with stereotactic core-needle biopsy (SCNB) change at follow-up mammography and to determine the optimal follow-up strategy and the delayed false-negative rate. MATERIALS AND METHODS From July 1992 through December 1995, 355 of 540 cases (66%) in which SCNB yielded benign results were managed with follow-up mammography. Mammographic follow-up was available for 298 of these cases (84%). Follow-up mammography reports were reviewed. When a change was reported, pre- and postbiopsy mammograms, pathology reports, and results of subsequent mammographic follow-up were reviewed. RESULTS Mammographic change occurred in 21 of 298 cases (7%) at intervals of 6-55 months (mean, 20 months). Change occurred after initial mammographic stability in 10 of 21 cases. Repeat biopsy was performed in 18 of 21 cases. Malignancy was diagnosed in two cases: one mass that changed at 6 months and one case of microcalcifications that changed at 24 months. This represented a delayed false-negative rate of 2% (two of 105 malignancies among 540 biopsies). CONCLUSION A small percentage of cases diagnosed as benign with SCNB will change on follow-up mammograms, which may necessitate repeat biopsy. These results suggest that 6-month follow-up for cases that yield nonspecific benign results at SCNB and yearly screening mammography for cases with specific benign results is a reasonable management strategy.
TNF-α Drives Human CD14+ Monocytes to Differentiate into CD70+ Dendritic Cells Evoking Th1 and Th17 Responses

Many mechanisms involving TNF-α, Th1 responses, and Th17 responses are implicated in chronic inflammatory autoimmune disease. Recently, the clinical impact of anti-TNF therapy on disease progression has resulted in re-evaluation of the central role of this cytokine and engendered novel concept of TNF-dependent immunity. However, the overall relationship of TNF-α to pathogenesis is unclear. Here, we demonstrate a TNF-dependent differentiation pathway of dendritic cells (DC) evoking Th1 and Th17 responses. CD14+ monocytes cultured in the presence of TNF-α and GM-CSF converted to CD14+ CD1alow adherent cells with little capacity to stimulate T cells. On stimulation by LPS, however, they produced high levels of TNF-α, matrix metalloproteinase (MMP)-9, and IL-23 and differentiated either into mature DC or activated macrophages (Mφ). The mature DC (CD83+ CD70+ HLA-DR high CD14low) expressed high levels of mRNA for IL-6, IL-15, and IL-23, induced naive CD4 T cells to produce IFN-γ and TNF-α, and stimulated resting CD4 T cells to secret IL-17. Intriguingly, TNF-α added to the monocyte culture medium determined the magnitude of LPS-induced maturation and the functions of the derived DC. In contrast, the Mφ (CD14highCD70+CD83−HLA-DR−) produced large amounts of MMP-9 and TNF-α without exogenous TNF stimulation. These results suggest that the TNF priming of monocytes controls Th1 and Th17 responses induced by mature DC, but not inflammation induced by activated Mφ. Therefore, additional stimulation of monocytes with TNF-α may facilitate TNF-dependent adaptive immunity together with GM-CSF-stimulated Mφ-mediated innate immunity.
Zinc and cadmium body burdens in terrestrial oligochaetes: Use and significance in environmental risk assessment

Uptake and elimination kinetics of zinc and cadmium were studied in the oligochaetes Enchytraeus albidus and Eisenia fetida. Even at the highest exposure concentrations where E. fetida survived, the internal zinc concentration was still regulated to a constant level. Enchytraeus albidus could not regulate the internal zinc concentration as well, and the body burden increased after exposure to high zinc concentrations. After transfer to clean soil, the internal zinc concentration dropped back to the control level within a few weeks. For both species, internal cadmium concentrations did not reach equilibrium during the uptake experiments. The internal concentrations causing 50% reduction in cocoon production for E. fetida exposed to cadmium varied between different soil types, indicating that no fixed critical body burdens exist. For both zinc and cadmium, bioaccumulation factors decreased with increasing soil metal concentrations. Bioaccumulation factors may therefore be poor indicators of environmental risk. Their dependence on the total soil concentration makes bioaccumulation factors also unsuitable for assessing the influence of soil characteristics on the bioavailability of metals in contaminated field soils. For the same reason, uptake rate constants are probably not suited for this purpose.
The College of Emergency Medicine curriculum: is the specialty-specific knowledge section fit for purpose?

Objective: To determine whether the knowledge specified in the specialty-specific section of the College of Emergency Medicine curriculum covered the diagnoses presenting to a UK teaching hospital emergency department. Method: An audit of 1000 sets of notes was undertaken, the diagnosis abstracted and mapped to the curriculum. Results: 1076 diagnoses were derived and all were covered by the curriculum. The three most common diagnostic categories were musculoskeletal, wound management and cardiology. Conclusion: The curriculum covered all the diagnoses in this sample. Knowing the frequency of a diagnosis could be used to inform training and assessment.
Multi-level virtual ring: An architecture for content routing in wireless sensor network

Two main problems prevent the deployment of content delivery in a wireless sensor network: the address, which is widely used in the Internet as the identifier, is meaningless in wireless network, and the routing efficiency is a big concern in wireless sensor network. This paper presents an embedded multi-level ring (MVR) structure to address those two problems. The MVR uses names rather than addresses to identify sensor nodes. The MVR routes packets on the name identifiers without being aware the location. Some sensor nodes are selected as the backbone nodes and are placed on the different levels of the virtual rings. MVR hashes nodes and contents identifiers, and stores them at the backbone nodes. MVR takes the cross-level routing to improve the routing efficiency. Further, MVR is constructed decentralized and runs on the mobile nodes themselves, requiring no central control. Experiments using ns2 simulator for up to 200 nodes show that the storage and bandwidth requirements of MVR grow slowly with the size of the network. Furthermore, MVR has demonstrated as self-administrating, fault-tolerant, and resilient under the different workloads. We also discuss alternative implementation options, and future work.
Sensory neuronal phenotype in galanin receptor 2 knockout mice: focus on dorsal root ganglion neurone development and pain behaviour

Galanin is a 29‐amino‐acid peptide expressed in dorsal root ganglion (DRG) neurones and spinal dorsal horn neurones. It affects pain threshold and has developmental and trophic effects. Galanin acts at three G‐protein‐coupled receptors, galanin receptors (GalR1–3), each expressed in the DRGs as suggested by in situ hybridization and/or reverse transcriptase‐polymerase chain reaction. The GalR2 knockout (–/–) mice permit studies on the contributions of this receptor subtype to the role of galanin at the spinal level. At 1 week after sciatic nerve transection (axotomy), there were 16–20% fewer neurones in intact and contralateral DRGs of –/– mice as compared with wild‐type (WT) mice. In addition, a significant neurone loss (26% reduction) was found in the ipsilateral DRGs of WT mice, whereas no further neurone loss was seen in –/– mice. Expression of several peptides has been examined after axotomy, including galanin, neuropeptide Y and two of its receptors as well as substance P, and no significant differences were found between –/– and WT mice in either ipsi‐ or contralateral DRGs, respectively. After thermal injury and spinal nerve ligation, onset and duration of hyperalgesia in the injured paw were similar in GalR2–/– and WT animals. Recovery from spinal nerve ligation‐caused allodynia had the same kinetics in –/– and WT animals. These data are in line with earlier observations from the peripheral and central nervous system, suggesting that galanin actions mediated by GalR2 subtype are of importance in neurodevelopment and neuroprotection.
Deposited in DRO : 11 October 2012 Version of attached le : Peer-review status of attached

An emerging trend is what has become commonly known as ‘Medical Tourism’ where patients travel to overseas destinations for specialised surgical treatments and other forms of medical care. With the rise of more affordable cross-border travel and rapid technological developments these movements are becoming more commonplace. A key driver is the platform provided by the internet for gaining access to healthcare information and advertising. There has been relatively little attention given to the role and impact of web-based information to inform Medical Tourism decisions. This article provides a brief overview of the most recent development in Medical Tourism and examines how this is linked to the emergence of specialized internet web sites. It produces a summary of the functionality of medical tourist sites, and situates Medical Tourism informatics within the broader literatures relating to information search, information quality and decision-making. This paper is both a call to strengthen the empirical evidence in this area, and also to advocate integrating Medical Tourism research within a broader conceptual framework.
Enrichment and Differentiation of Human Germ‐Like Cells Mediated by Feeder Cells and Basic Fibroblast Growth Factor Signaling

Human embryonic stem cells (hESCs) have recently demonstrated the potential for differentiation into germ‐like cells in vitro. This provides a novel model for understanding human germ cell development and human infertility. Mouse embryonic fibroblast (MEF) feeders and basic fibroblast growth factor (bFGF) are two sources of signaling that are essential for primary culture of germ cells, yet their role has not been examined in the derivation of germ‐like cells from hESCs. Here protein and gene expression demonstrated that both MEF feeders and bFGF can significantly enrich germ cell differentiation from hESCs. Under enriched differentiation conditions, flow cytometry analysis proved 69% of cells to be positive for DDX4 and POU5F1 protein expression, consistent with the germ cell lineage. Importantly, removal of bFGF from feeder‐free cultures resulted in a 50% decrease in POU5F1‐ and DDX4‐positive cells. Quantitative reverse transcription‐polymerase chain reaction analysis established that bFGF signaling resulted in an upregulation of genes involved in germ cell differentiation with or without feeders; however, feeder conditions caused significant upregulation of premigratory/migratory (Ifitm3, DAZL, NANOG, and POU5F1) and postmigratory (PIWIL2, PUM2) genes, along with the meiotic markers SYCP3 and MLH1. After further differentiation, >90% of cells expressed the meiotic proteins SYCP3 and MLH1. This is the first demonstration that signaling from MEF feeders and bFGF can induce a highly enriched population of germ‐like cells derived from hESCs, thus providing a critically needed model for further investigation of human germ cell development and signaling.
EFFECT OF ANTISENSE OLIGONUCLEOTIDES TO NUCLEAR FACTOR-κB ON THE SURVIVAL OF LPS-INDUCED ARDS IN MOUSE

Because nuclear factor (NF)- κ B-regulated cytokines, including tumor necrosis factor- α (TNF- α) , from monocytes and macrophages have been implicated in the pathogenesis and development of septic shock and acute respiratory distress syndrome (ARDS), the effect of the antisense oligonucleotide to the p65 subunit of NF- κ B on the survival of lipopolysaccharide (LPS)-induced ARDS in BALB/c mice was examined. None and 70% of the animals died of diffuse hemorrhagic lung edema 1 to 2.5 days after intraperitoneal administration of 10 and 20 mg/kg LPS alone, respectively. Intravenously administered antisense oligonucleotide alone did not produce any significant changes in the behavior or lung histology. After intravenous administration of the anti-sense oligonucleotide, both peripheral blood monocytes and alveolar macrophages in bronchoalveolar lavage fluid were confirmed to contain sufficiently large amounts of intracellular antisense oligonucleotides for their function using fluorescein isothiocyanate (FITC)-labeled microscopy. The antisense oligonucleotide administered 6 hours before the intraperitoneal administration of LPS significantly decreased the survival rate with the progress of hemorrhagic edema in lung histology; 90% and 100% of animals treated with the antisense oligonuleotide died 0.5 to 1.5 days after the administration of 10 and 20 mg/kg LPS, respectively. These findings suggest that the suppression of cytokines and mediators in monocytes and alveolar macrophages by the antisense oligonucleotide to the p65 subunit of NF- κ B worsens the survival of LPS-induced ARDS in mice with the progress of hemorrhagic lung edema.