Implications of the innate immune response to adenovirus and adenoviral vectors

Adenovirus (AdV) is a common cause of respiratory illness in both children and adults. Respiratory symptoms can range from those of the common cold to severe pneumonia. Infection can also cause significant disease in the immunocompromised and among immunocompetent subjects in close quarters. Fortunately, infection with AdV in the normal host is generally mild. This is one reason why its initial use as a gene-therapy vector appeared to be so promising. Unfortunately, both innate and adaptive responses to the virus have limited the development of AdV vectors as a tool of gene therapy by increasing toxicity and limiting duration of transgene expression. This article will focus on the innate immune response to infection with wild-type AdV and exposure to AdV gene-therapy vectors. As much of the known information relates to the pulmonary inflammatory response, this organ system will be emphasized. This article will also discuss how that understanding has led to the creation of new vectors for use in gene therapy.     

Copper nanoparticles: Green synthesis and managing fruit rot disease of chilli caused by Colletotrichum capsici

The present study was focused on synthesis and characterization of copper nanoparticles to evaluate their efficacy against fruit rot pathogen of chilli crop. The green synthesis of nanoparticles was carried out by using extracts of Eucalyptus and Mint leaves. The synthesis of copper nanoparticles was confirmed by XRD, PSA, SEM and TEM. The average size of these particles synthesized by Eucalyptus leaf extract (CuNP-E) ranged from 10 to 130 nm, while as size of Mint leaf extract synthesized particles (CuNP-M) ranged from 23 to 39 nm, thus confirming their nano size. These green synthesized copper nanoparticles were evaluated against Colletotrichum capsici where Carbendazim 50 WP @ 500 ppm and copper oxychloride 50 WP @ 2500 ppm served as standard checks. The mycelia inhibition of Colletotrichum capsici caused by copper nanoparticles was studied on PDA medium. CuNP-M @ 1000 ppm showed highest mycelial inhibition of 99.78% followed by 93.75% at 500 ppm and CuNP-E @ 1000 ppm compared to standard fungicides, carbendazim 50 WP @ 500 ppm (72.82%), and copper oxychloride 50 WP @ 2500 ppm (85.85%). The CuNP-M @ 500 ppm were significantly superior to carbendazim 50 WP @ 500 ppm and copper oxychloride 50 WP @ 2500 ppm, but was statistically at par with CuNP-E @ 1000 ppm. This shows effectiveness of much lower concentration of copper nanoparticles compared to conventional fungicides. In detached fruit method, nanoparticles applied before inoculation of pathogen showed better results with regard to incubation period, lesion number and lesion size than after inoculation of pathogen. The present study reveals a simple, convenient, non-toxic and cost-efficient technique for the synthesis of nanoparticles and their effectiveness against Colletotrichum capsici. CuNP-M first time synthesized and evaluated against Colletotrichum capsici performed better than CuNP-E.     

Label free high throughput screening for apoptosis inducing chemicals using time-lapse microscopy signal processing

Label free time-lapse microscopy has opened a new avenue to the study of time evolving events in living cells. When combined with automated image analysis it provides a powerful tool that enables automated large-scale spatiotemporal quantification at the cell population level. Very few attempts, however, have been reported regarding the design of image analysis algorithms dedicated to the detection of apoptotic cells in such time-lapse microscopy images. In particular, none of the reported attempts is based on sufficiently fast signal processing algorithms to enable large-scale detection of apoptosis within hours/days without access to high-end computers. Here we show that it is indeed possible to successfully detect chemically induced apoptosis by applying a two-dimensional linear matched filter tailored to the detection of objects with the typical features of an apoptotic cell in phase-contrast images. First a set of recorded computational detections of apoptosis was validated by comparison with apoptosis specific caspase activity readouts obtained via a fluorescence based assay. Then a large screen encompassing 2,866 drug like compounds was performed using the human colorectal carcinoma cell line HCT116. In addition to many well known inducers (positive controls) the screening resulted in the detection of two compounds here reported for the first time to induce apoptosis.