Correction of deletions in mammalian cells by gene conversion

We have constructed substrates to study the conversion of deletions in mammalian cells both extrachromosomally and after the stable integration of the substrates into the chromosome. These substrates were designed to study gene conversion without the complication of reciprocal recombination events. The substrates contain insertion or deletion mutations of the neomycin resistance gene (neo)and an internal, homologous fragment of the neo gene neo-526,such that gene conversion from neo-526to the mutated neogene restores a functional neogene. We have shown that extrachromosomally insertions of 10 bp or deletions of 22 or 167 bp are converted to wild-type at similar frequencies (1–6 × 10^–4 Chromosomal gene conversion occurred at frequencies of about 10^–6–10^–7 per cell generation. As expected from the experimental design, all recombination events analyzed in mammalian cells using these substrates appear to be due to gene conversion.     

A Rare Association of Left Ventricular Diverticulum and Noncompaction: Continuum of Common Developmental Abnormality?

A previously healthy 16‐year‐old girl presented with history of fever of 3 months duration without any significant clinical finding on examination. Transthoracic echocardiography (TTE) revealed moderate pericardial effusion and later diagnosed as tubercular in etiology. Besides, TTE also showed an incidental finding of large left ventricular (LV) diverticulum in submitral region along the postero‐basal segment and left ventricular noncompaction (LVNC). Nonapical LV diverticulum is a rare variant of this uncommon congenital condition and its association with LVNC has not been reported in literature. Patient was discharged on antitubercular therapy for pericardial effusion and advised for 6 monthly follow‐up for diverticulum.     

Association of initial tuberculin sensitivity, age and sex with the incidence of tuberculosis in south India: a 15-year follow-up.

OBJECTIVE: To determine the association of initial tuberculin sensitivity, age and sex with the development of tuberculosis. METHODS: A 15-year follow-up of 280000 subjects in south India, where new cases of tuberculosis were detected mainly by periodic population surveys. Life-table technique was employed to estimate tuberculosis incidence and disease risk in survivors. The independent effect of tuberculin sensitivity, sex and age at intake was determined using Cox's proportional hazard model. RESULTS: Taking subjects with reaction size 0-7 mm to 3 IU PPD-S as reference group, the adjusted relative risk (RR) for developing culture-positive tuberculosis was 1.1, 1.9, 2.9, 3.6 and 3.3 for those with indurations of 8-11, 12-15, 16-19, 20-24 and > or = 25 mm (P < 0.01). Considering subjects aged 0-4 years as reference group, the adjusted RR for the other groups increased from 1.7 to 10.8 (P < 0.01). Males had a substantially higher incidence (adjusted RR 3.0, P < 0.001). The risk of culture-positive tuberculosis over 15 years in survivors was 3.3% (5.0% in males and 1.6% in females), and increased substantially with tuberculin sensitivity at intake. In those with > or = 12 mm at intake, the approximate lifetime risk was 6.1% (8.6% in males and 3.1% in females). CONCLUSION: The incidence of tuberculosis increased steadily with tuberculin sensitivity to PPD-S and age at intake. Males had a significantly higher risk than females in every PPD-S group and the overall risk was three-fold higher.     

Disorder in Ho2Ti2−xZrxO7: pyrochlore to defect fluorite solid solution series

Pyrochlore (A₂B₂O₇) is an important, isometric structure-type because of its large variety of compositions and structural derivatives that are generally related to different disordering mechanisms at various spatial scales. The disordering is key to understanding variations in properties, such as magnetic behavior or ionic conduction. Neutron and X-ray total scattering methods were used to investigate the degree of structural disorder in the Ho₂Ti_2−xZr_xO₇ (x = 0.0–2.0, Δx = 0.25) solid solution series as a function of the Zr-content, x. Ordered pyrochlores (Fd$\bar{3}$m) disorder to defect fluorite (Fm$\bar{3}$m) via cation and anion disordering. Total scattering experiments with sensitivity to the cation and anion sublattices provide unique insight into the underlying atomic processes. Using simultaneous Rietveld refinement (long-range structure) and small-box refinement PDF analysis (short-range structure), we show that the series undergoes a rapid transformation from pyrochlore to defect fluorite at x ≈ 1.2, while the short-range structure exhibits a linear increase in a local weberite-type phase, C222₁, over the entire composition range. Enthalpies of formation from the oxides determined using high temperature oxide melt solution calorimetry support the structural data and provide insight into the effect of local ordering on the energetics of disorder. The measured enthalpies of mixing are negative and are fit by a regular solution parameter of W = −31.8 ± 3.7 kJ mol⁻¹. However, the extensive short-range ordering determined from the structural analysis strongly suggests that the entropies of mixing must be far less positive than implied by the random mixing of a regular solution. We propose a local disordering scheme involving the pyrochlore 48f to 8a site oxygen Frenkel defect that creates 7-coordinated Zr sites contained within local weberite-type coherent nanodomains. Thus, the solid solution is best described as a mixture of two phases, with the weberite-type nanodomains triggering the long-range structural transformation to defect fluorite after accumulation above a critical concentration (50% Ti replaced by Zr).

Combined neutron and X-ray total scattering with calorimetric measurements of the solid solution series Ho₂Ti_2−xZr_xO₇ reveals a complex order–disorder transition across short, intermediate, and long length scales induced by chemical substitution.     

Molecular mechanisms regulating CD13‐mediated adhesion

CD13/Aminopeptidase N is a transmembrane metalloproteinase that is expressed in many tissues where it regulates various cellular functions. In inflammation, CD13 is expressed on myeloid cells, is up‐regulated on endothelial cells at sites of inflammation and mediates monocyte/endothelial adhesion by homotypic interactions. In animal models the lack of CD13 alters the profiles of infiltrating inflammatory cells at sites of ischaemic injury. Here, we found that CD13 expression is enriched specifically on the pro‐inflammatory subset of monocytes, suggesting that CD13 may regulate trafficking and function of specific subsets of immune cells. To further dissect the mechanisms regulating CD13‐dependent trafficking we used the murine model of thioglycollate‐induced sterile peritonitis. Peritoneal monocytes, macrophages and dendritic cells were significantly decreased in inflammatory exudates from global CD13^KO animals when compared with wild‐type controls. Furthermore, adoptive transfer of wild‐type and CD13^KO primary myeloid cells, or wild‐type myeloid cells pre‐treated with CD13‐blocking antibodies into thioglycollate‐challenged wild‐type recipients demonstrated fewer CD13^KO or treated cells in the lavage, suggesting that CD13 expression confers a competitive advantage in trafficking. Similarly, both wild‐type and CD13^KO cells were reduced in infiltrates in CD13^KO recipients, confirming that both monocytic and endothelial CD13 contribute to trafficking. Finally, murine monocyte cell lines expressing mouse/human chimeric CD13 molecules demonstrated that the C‐terminal domain of the protein mediates CD13 adhesion. Therefore, this work verifies that the altered inflammatory trafficking in CD13^KO mice is the result of aberrant myeloid cell subset trafficking and further defines the molecular mechanisms underlying this regulation.     

Hydrothermal synthesis of cobalt telluride nanorods for a high performance hybrid asymmetric supercapacitor

Cobalt telluride nanostructured materials have demonstrated various applications, particularly in energy generation and storage. A high temperature and reducing atmosphere are required for the preparation of cobalt telluride-based materials, which makes this a difficult and expensive process. The development of a facile route for producing the desirable nanostructure of cobalt telluride remains a great challenge. We demonstrated a simple hydrothermal method for preparing cobalt telluride nanorods (CoTe NRs) and telluride nanorods (Te NRs) for supercapacitor applications. The morphology of CoTe NRs and Te NRs was analyzed using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The prepared CoTe NR electrode material exhibited a high specific capacity of 170 C g⁻¹ at a current density of 0.5 A g⁻¹ with an exceptional cyclic stability. The asymmetric supercapacitor was assembled using CoTe NRs and orange peel-derived activated carbon (OPAA-700) as a positive and negative electrode, respectively. The fabricated device delivered a high energy density of 40.7 W h kg⁻¹ with a power density of 800 W kg⁻¹ at 1 A g⁻¹ current density. When the current density was increased to 30 A g⁻¹, the fabricated device delivered a high power density of 22.5 kW kg⁻¹ with an energy density of 16.3 W h kg⁻¹. The fabricated asymmetric supercapacitor displayed a good cyclic stability performance for 10 000 cycles at a high current density of 30 A g⁻¹ and retained 85% of its initial capacity for after 10 000 cycles. The prepared materials indicate their applicability for high performance energy storage devices.

A one-step hydrothermal derived cobalt telluride nanorods and activated carbon-based hybrid asymmetric supercapacitor delivered a high energy (40.7 W h kg⁻¹) and power density (22.5 kW kg⁻¹) with an electrochemical stability of 85% for 10000 cycles.     

Immunomodulatory Effects of Newcastle Disease Virus AF2240 Strain on Human Peripheral Blood Mononuclear Cells

Immunotherapy has raised the attention of many scientists because it hold promise to be an attractive therapeutic strategy to treat a number of disorders. In this study, the immunomodulatory effects of low titers of Newcastle disease virus (NDV) AF2240 on human peripheral blood mononuclear cells (PBMC) were analyzed. We evaluated cytokine secretion and PBMC activation by cell proliferation assay, immunophenotyping and enzyme linked immunosorbent assay. The proliferation of the human PBMC was measured to be 28.5% and 36.5% upon treatment with 8 hemaglutinin unit (HAU) and 2 HAU of NDV respectively. Interestingly, the percentage of cells with activating markers CD16 and CD56 were increased significantly. Furthermore, the intracellular perforin and granzyme levels were also increased upon virus infection. Human PBMC treated with NDV titer 8 HAU was found to stimulate the highest level of cytokine production including interferon-γ, interleukin-2 and interleukin-12. The release of these proteins contributes to the antitumor effect of PBMC against MCF-7 breast cancer cells. Based on the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay, activated human PBMC showed high cytolytic efficiency towards human breast tumor cells. In summary, NDV was able to stimulate PBMC proliferation, cytokine secretion and cytolytic activity.     

Effect of SiC Nanoparticles on AZ31 Magnesium Alloy

Magnesium alloys are attractive for the production of lightweight parts in modern automobile and aerospace industries due to their advanced properties. Their mechanical properties are usually enhanced by the incorporation with reinforcement particles. In the current study, reinforced AZ31 magnesium alloy was fabricated through the addition of bulk Al and the incorporation of SiC nanoparticles using a stir casting process to obtain AZ31-SiC nanocomposites. Scanning electron microscope (SEM) investigations revealed the formation of Mg₁₇Al₁₂ lamellar intermetallic structures and SiC clusters in the nanocomposites. Energy dispersive spectroscopy (EDS) detected the uniform distribution of SiC nanoparticles in the AZ31-SiC nanocomposites. Enhancements in hardness and yield strength (YS) were detected in the fabricated nanocomposites. This behavior was referred to a joint strengthening mechanisms which showed matrix-reinforcement coefficient of thermal expansion (CTE) and elastic modulus mismatches, Orowan strengthening, and load transfer mechanism. The mechanical properties and wear resistance were gradually increased with an increase in SiC content in the nanocomposite. The maximum values were obtained from nanocomposites containing 1 wt% of SiC (AZ31-1SiC). AZ31-1SiC nanocomposite YS and hardness were improved by 27% and 30%, respectively, compared to AZ31 alloy. This nanocomposite also exhibited the highest wear resistance; its wear mass loss and depth of the worn surface decreased by 26% and 15%, respectively, compared to AZ31 alloy.     

Positive modulation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-type glutamate receptors elicits neuroprotection after trimethyltin exposure in hippocampus

Experiments were designed to determine if gender influences the cardiac toxicity elicited by chronic high-level ethanol intake. Male and female ethanol-preferring P-rats were allowed free access to drinking water or water containing 25% ethanol for 6 months. Left atrial preparations were then isolated, bathed in Krebs-Henseleit solution (37 degrees C), and used to examine basal contractility at 3.0 Hz stimulation, the force-frequency relationship, and the positive inotropic response to the beta-adrenoceptor agonist isoproterenol. Basal contractile function was not affected significantly by ethanol in either gender; however, atria from ethanol-treated male rats displayed diminished contractility compared to control males when measured at slow stimulation frequencies (0.1 and 0.2 Hz), during post rest potentiation (at rest intervals of 20-60 s), and in response to higher concentrations of isoproterenol (> or =3 x 10(-7) M; EC50 values were not affected). In contrast, female atria showed no effect of chronic ethanol consumption. These data suggest that ethanol consumption diminishes the cardiac reserve in male, but not female rats.