Genome-wide transcription analysis of interaction between the human macrophage and Mycobacterium tuberculosis during concurrent drug administration by conventional and novel methods

Targeted drug delivery to alveolar macrophages harboring Mycobacterium tuberculosis (Mtb) holds promise of high efficacy against pulmonary tuberculosis (TB). It was investigated whether inhalable microparticles (MP) can rescue macrophages from ‘alternative’ activation induced by pathogenic Mtb in addition to achieving targeted drug delivery. A genome-wide transcription analysis (Affymetrix HG-U133 Plus 2.0 DNA microarray) of THP-1 cell line derived macrophages was undertaken after exposing them to infection with 10 MOI of MTB H37Rv at 0, 12 and 24 hours post infection. The Molecular markers of macrophage bactericidal activity were assayed in THP-1- and primary human peripheral blood mononuclear cell (PBMC)-derived macrophages, in the presence or absence of soluble anti-tuberculosis drugs, drug-containing MP and blank MP. About 1,500 genes were differentially upregulated and about 500 genes differentially downregulated in response to various modes of treatment. Variations were also observed in the kinetics of gene expression. Cluster analysis indicated activation of several pathways related to innate immune response (cytokines, chemokines, receptors and ligands), apoptosis, cytoskeleton and membrane remodeling, general metabolism and general housekeeping. Some of these results were validated at the functional level, by studying caspase activities, concentrations and time-courses of effector molecules , rates/extents of apoptosis and nitrite oxide induction. Production of cytokines and NO, apoptosis, and bacterial survival were studied as pharmacodynamic outcomes. Cytokine responses of THP-1 derived macrophages were estimated. MP reversed suppression of tumor necrosis factor (TNF) induced by infection, and transiently upregulated γ-interferon (IFN-γ). Drug-free MP surprisingly induced IFN-γ, but not TNF. Primary cells responded to MP, regardless of drug content, by upregulation of NO; but THP-1-derived cells did not respond to blank MP. About 19% of infected cells exposed to MP underwent apoptosis as compared to ~11% cells treated with soluble drugs or blank MP. Cell death induced by blank MP was caspase-independent. Only drug-containing MP induced apoptosis through caspase-8 and caspase-9. Bacterial survival after different treatments varied between individuals. In the best case, while untreated infection resulted in survival of 900±141 colony forming units (CFU), treatment with soluble drugs, drug-containing MP and blank MP respectively, reduced CFU counts to 8.5± 0.7, 3±1.4 and 102±138.6. The results suggest a role of the drug delivery system in macrophage activation as a component of therapeutic strategy against TB

Contribution of rankl regulation to bone resorption induced by PTH receptor activation in osteocytes

Indiana University-Purdue University Indianapolis (IUPUI)PTH increases osteoclasts by upregulating RANKL in cells of the osteoblastic lineage, but the precise differentiation stage of the PTH target cell remains undefined. Recent findings demonstrate that PTH regulates gene expression in osteocytes and that these cells are an important source of RANKL. We therefore investigated whether direct regulation of the RANKL gene by PTH in osteocytes is required to stimulate osteoclastic bone resorption. To address this question, we examined bone resorption and RANKL expression in transgenic mice in which PTH receptor signaling is activated only in osteocytes (DMP1-caPTHR1) crossed with mice lacking the distal control region regulated by PTH in the RANKL gene (DCR -/-). Longitudinal analysis of circulating C-terminal telopeptide (CTX) in male mice showed elevated resorption in growing mice that progressively decreased to plateau at 3-5 month of age. Resorption was significantly higher (~100%) in DMP1-caPTHR1 mice and non-significantly lower (15-30%) in DCR -/-mice, versus wild type littermates (WT) across all ages. CTX in compound DMP1-caPTHR1; DCR -/-mice was similar to DMP1-caPTHR1 mice at 1 and 2 months of age, but by 3 months of age, was significantly lower compared to DMP1-caPTHR1 mice (50% higher than WT), and by 5 months, it was undistinguishable from WT mice. Micro-CT analysis revealed lower tissue material density in the distal femur of DMP1-caPTHR1 mice, indicative of high remodeling, and this effect was partially corrected in compound vi mice. The increased resorption exhibited by DMP1-caPTHR1 mice was accompanied by elevated RANKL mRNA in bone at 1 and 5 months of age. RANKL expression levels displayed similar patterns to CTX levels in DMP1-caPTHR1; DCR -/-compound mice at 1 and 5 month of age. The same pattern of expression was observed for M-CSF. We conclude that resorption induced by PTH receptor signaling requires direct regulation of the RANKL gene in osteocytes, but this dependence is age specific. Whereas DCR-independent mechanisms involving gp130 cytokines or vitamin D 3 might operate in the growing skeleton, DCR-dependent, cAMP/PKA/CREB-activated mechanisms mediate resorption induced by PTH receptor signaling in the adult skeleton

PLA Microparticles for Pulmonary Delivery of AntiTB drugs: Biodistribution study

A dry powder inhalable (DPI) microparticles comprising anti-tuberculosis drugs incorporated in biodegradable polymers was developed for the treatment of pulmonary tuberculosis (P. Muttil _et al_. 2007). Poly L-lactic acid (PLA) microparticles incorporating a high payload of rifabutin and isoniazid were fabricated by spray drying (Buchi 190). Microparticles were composed of PLA and the drugs (rifabutin and isoniazid) at a 2:1:1 weight ratio. Microparticles of desired high encapsulation efficiency and sustained release characteristics were produced having a diameter range of 2-10 µm (Malvern Mastersizer 2000). Differential scanning calorimetry (DSC) was carried out to study drug polymer interaction. The time course of tissue biodistribution following a single inhalation dose of microparticles was evaluated. 
Thirty-two BALB/c mice were divided into groups of four and administered the DPI using an in-house (nose only) apparatus (Kaur _et al_. 2008; Verma _et al_. 2008). A validated HPLC method was used for determination of rifabutin and isoniazid in the lungs (target organ), liver and kidneys (major sites of toxicity) at different time-points after inhalation. A comparison was made with mice receiving free drugs (intravenous) at equivalent doses. Deposition of microparticles in lungs of mice following aerosolization was also evaluated. Pharmacokinetic parameters in different organs were calculated using WinNonlin software version 5.2. Area under the concentration-time curve observed (AUC~obs~), C~max~, half-life (t~½~) and clearance (CL) in lungs following inhalation /intravenous administration were:
*Rifabutin*: AUC~obs~-96h= 1697.39 ±154.67 (187.63 ±23.93) µg/ml^-1^hr^-1^; C~max~ = 33.42±3.80 (4.17±0.31) µg.ml^-1^; t~½~= 78.08±9.42 (34.00 ±3.31) and Cl= 1.16±.22 (0.68 ±0.45) ml.h^-1^.
*Isoniazid*: AUC~obs~-24h= 566.31±123.96 (99.85 ±14.24) µg/ml^-1^hr^-1^; Cmax= 24.02±1.71 (8.16±0.93) µg.ml^-1^; t~½~= 25.88±12.16 (6.45±3.24) h; and Cl= 5.47±1.30 (0.96±0.14) ml.h^-1^.
The relative bioavailability of both drugs incorporated in microparticles was significantly higher compared with free drugs. Peak levels of isoniazid and rifabutin in lungs (target organ) were much higher than those in the liver and kidney of mice in case of inhalation as compared to intravenous administration. Inhalation of microparticles resulted in targeting both drugs to the lungs, with the effect being more pronounced in the case of rifabutin than isoniazid. High and prolonged drug concentrations and increased AUC values (~9-fold and ~6 fold increase of rifabutin and isoniazid in case of lungs) with respect to free drugs were observed. Significant decrease in drug concentration was found in the liver and kidneys. Drug levels were maintained above the minimum inhibitory concentration (MIC) in organs through out the study after administration of encapsulated drugs. Based on favorable biodistribution kinetics, these microparticles hold great potential in reducing dosing frequency and toxicity of antituberculosis drugs.
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Perspective of buried oxide thickness variation on triple metal-gate (TMG) recessed-S/D FD-SOI MOSFET

Recently, Fully-Depleted Silicon on Insulator (FD-SOI) MOSFETs have been accepted as a favourable technology beyond nanometer nodes, and the technique of Recessed-Source/Drain (Re-S/D) has made it more immune in regards of various performance factors. However, the proper selection of Buried-Oxide (BOX) thickness is one of the major challenges in the design of FD-SOI based MOS devices in order to suppress the drain electric penetrations across the BOX interface efficiently. In this work, the effect of BOX thickness on the performance of TMG Re-S/D FD-SOI MOSFET has been presented at 60 nm gate length. The perspective of BOX thickness variation has been analysed on the basis of its surface potential profile and the extraction of the threshold voltage by performing two-dimensional numerical simulations. Moreover, to verify the short channel immunity, the impact of gate length scaling has also been discussed. It is found that the device attains two step-up potential profile with suppressed short channel effects. The outcomes reveal that the Drain Induced Barrier Lowering (DIBL) values are lower among conventional SOI MOSFETs. The device has been designed and simulated by using 2D numerical ATLAS Silvaco TCAD simulator

Early recurrent ischemic stroke complicating intravenous thrombolysis for stroke: incidence and association with atrial fibrillation

<p><b>Background and Purpose:</b> Mechanisms of early neurologic deterioration after treatment with intravenous, recombinant, tissue-type plasminogen activator (IV rt-PA) include symptomatic intracerebral hemorrhage (SICH) and early recurrent ischemic stroke. We observed a number of cases of acute deterioration due to recurrent ischemic events.</p> <p><b>Methods:</b> We undertook a single-center, retrospective analysis of consecutive acute stroke patients treated with IV rt-PA between January 2006 and December 2008 to define the incidence of early neurologic deterioration (>= 4-point drop on the National Institutes of Health Stroke Scale within 72 hours) and its mechanism. Deterioration was attributed to SICH when associated with a PH1 or PH2 hemorrhage on postdeterioration computed tomography scans, to recurrent ischemic stroke when there was clinical and radiologic evidence of a new territorial infarction or new vessel occlusion, and otherwise to evolution of the incident stroke.</p> <p><b>Results:</b> Of 228 consecutive IV rt-PA-treated patients, 34 (15%) developed early neurologic deterioration, 18 (8%) secondary to incident strokes 10 (4.4%) due to SICH, and 6 (2.6%) due to early recurrent ischemic events, which were significantly associated with atrial fibrillation (present in 5 of 6 patients; 4 paroxysmal, 1 permanent). In 4 patients, sudden clinical deterioration developed during or shortly after IV rt-PA infusion, and in 2, deterioration developed 3 days later. All died 2 days to 2 weeks later. The single case without atrial fibrillation had a recurrent, contralateral, middle cerebral artery stroke during IV rt-PA infusion and multiple high-signal emboli detected by transcranial Doppler. Early recurrent ischemic stroke accounted for 5 of 12 (42%) cases of early neurologic deterioration in patients with atrial fibrillation.</p> <p><b>Conclusion:</b> In this single-center series, the incidence of early recurrent ischemic stroke after IV rt-PA was 2.6% and was associated with previous atrial fibrillation.</p&gt

Selection and breeding programs for improving Indian sheep

Call number: LD2668 .R4 1967 S5

Parking guidance and authentication system

Whenever the number of parking lots became large the problems related to parking management became more complicated. The most two important problems that related to large parking places are: First, the long searching time for available lots, especially at the peak time. The second problem is the unauthorized-parking situations. Nowadays, there are parking guidance systems that guide the drivers toward the available lots. As a result, reduce the searching time for available lots, as well as reduce the congestion in that parking area. However, these systems do not solve the problem of unauthorized-parked vehicles. This study aims to combine along with guidance system, a mechanism to verify the legality of vehicles parked at authorized-parking lots. Therefore, this study will focus on two modules which are the detection module and the identification module. In the detection module, wireless sensors such as ultrasonic and y-axis magnetometers have proven their ability in detecting vehicles. And in the identification module, active RFID including both tag and reader are very suitable for this purpose. This project has significant implications for large institutions, by making the parking lots that allocated to their staff within the same customer parking area. In addition, it reduces the required manpower for managing that parking area

(Correcting) misdiagnoses of asthma: A cost effectiveness analysis

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Background: The prevalence of physician-diagnosed-asthma has risen over the past three decades and misdiagnosis of asthma is potentially common. Objective: to determine whether a secondary-screening-program to establish a correct diagnosis of asthma in those who report a physician diagnosis of asthma is cost effective.Method: Randomly selected physician-diagnosed-asthmatic subjects from 8 Canadian cities were studied with an extensive diagnostic algorithm to rule-in, or rule-out, a correct diagnosis of asthma. Subjects in whom the diagnosis of asthma was excluded were followed up for 6-months and data on asthma medications and heath care utilization was obtained. Economic analysis was performed to estimate the incremental lifetime costs associated with secondary screening of previously diagnosed asthmatic subjects. Analysis was from the perspective of the Canadian healthcare system and is reported in Canadian dollars.Results: Of 540 randomly selected patients with physician diagnosed asthma 150 (28%; 95%CI 19-37%) did not have asthma when objectively studied. 71% of these misdiagnosed patients were on some asthma medications. Incorporating the incremental cost of secondary-screening for the diagnosis of asthma, we found that the average cost savings per 100 individuals screened was $35,141 (95$4,588-$69,278).Conclusion: Cost savings primarily resulted from lifetime costs of medication use averted in those who had been misdiagnosed.This work was funded by the Canadian Institute of Health Research, Canada and the University Of Ottawa Division Of Respiratory Medicine

Enabling equitable collective action and policy change for poverty reduction and improved natural resource management in the Eastern African highlands:

"The role of local and external institutions in natural resource management (NRM) is gaining attention in the literature, fostering greater understanding of the relationship between collective action and poverty, collective action and equity, and the conditions under which collective action institutions take root. It has also led to increased understanding of how uncritical practices by external development institutions can propagate social inequities in NRM. Yet little research has been conducted to understand how to foster local collective action institutions where they are absent, or to improve institutional practice. This research integrates empirical and action research in an effort to generate ‘working solutions' to problems facing rural communities in their efforts to manage their natural resources in the highlands of Ethiopia and Uganda. Following a brief introduction to the literature and the research, findings are presented according to two distinct phases of research. Data are first presented on existing forms of collective action, the influence of local and external institutions on economic development, and NRM problems that persist despite their negative livelihood consequences. Action research themes selected from a list of identified problems are then presented in greater detail, with lessons learnt thus far in attempting to overcome institutional barriers to improved NRM. The paper concludes with a discussion of the implications of findings for research, institutional practice, and policy." authors' abstractNatural resource management, Collective action, Equity, Livelihoods, East African highlands, Gender, Environmental management,

Parathyroid hormone receptor signaling induces bone resorption in the adult skeleton by directly regulating the RANKL gene in osteocytes

PTH upregulates the expression of the receptor activator of nuclear factor κB ligand (Rankl) in cells of the osteoblastic lineage, but the precise differentiation stage of the PTH target cell responsible for RANKL-mediated stimulation of bone resorption remains undefined. We report that constitutive activation of PTH receptor signaling only in osteocytes in transgenic mice (DMP1-caPTHR1) was sufficient to increase Rankl expression and bone resorption. Resorption in DMP1-caPTHR1 mice crossed with mice lacking the distal control region regulated by PTH in the Rankl gene (DCR(-/-)) was similar to DMP1-caPTHR1 mice at 1 month of age, but progressively declined to reach values undistinguishable from wild-type (WT) mice at 5 months of age. Moreover, DMP1-caPTHR1 mice exhibited low tissue material density and increased serum alkaline phosphatase activity at 5 month of age, and these indices of high remodeling were partially and totally corrected in compound DMP1-caPTHR1;DCR(-/-) male mice, and less affected in female mice. Rankl expression in bones from DMP1-caPTHR1 mice was elevated at both 1 and 5 months of age, whereas it was high, similar to DMP1-caPTHR1 mice at 1 month, but low, similar to WT levels at 5 months in compound mice. Moreover, PTH increased Rankl and decreased Sost and Opg expression in ex vivo bone organ cultures established from WT mice, but only regulated Sost and Opg expression in cultures from DCR(-/-) mice. PTH also increased RANKL expression in osteocyte-containing primary cultures of calvarial cells, in isolated murine osteocytes, and in WT but not in DCR(-/-) osteocyte-enriched bones. Thus, PTH upregulates Rankl expression in osteocytes in vitro, ex vivo and in vivo, and resorption induced by PTH receptor signaling in the adult skeleton requires direct regulation of the Rankl gene in osteocytes.We thank Dr. Keith Condon and Ms. Naomie Olivos for technical assistance and Dr Munro Peacock for measurement of alkaline phosphatase. This research was supported by the National Institutes of Health (R01DK076007 and American Recovery and Reinvestment Act supplement S10-RR023710 to T.B.) and the Veterans Administration (Merit Review I01BX002104 to T.B.). Disclosure Summary: The authors declare that no conflict of interest exists