Detection of Pseudomonas fluorescens from broth, water and infected tissues by loop-mediated isothermal amplification (LAMP) method

Loop mediated isothermal amplification is rapid, highly sensitive and specifically developed method for detection of bacterial infections. AprX gene for alkaline metalloprotease of Pseudomonas fluorescens was used to design four primers and loop mediated isothermal amplification (LAMP) conditions were standardized for amplification of DNA. LAMP primers successfully amplified P. fluorescens from DNA and bacterial cells taken directly from broth, water and infected tissues with high specificity and sensitivity (10 pg) under isothermal condition at 61°C.Key words: Pseudomonas fluorescens, loop mediated isothermal amplification (LAMP), rapid, simple, specificity, sensitivity

Povećanje oslobađanja gliklazida iz smjesa dobivenih geometrijskim miješanjem

The poorly water soluble antidiabetic drug gliclazide was selected to study the effect of excipients on dissolution rate enhancement. Ordered mixtures of micronized gliclazide with lactose, mannitol, sorbitol, maltitol and sodium chloride were prepared by manual shaking of glass vials containing the drug and excipient(s). Different water soluble excipients, addition of surfactant and superdisintegrant, drug concentration and carrier particle size influenced the dissolution rate of the drug. Dissolution rate studies of the prepared ordered mixtures revealed an increase in drug dissolution with all water soluble excipients. The order of dissolution rate improvement for gliclazide was mannitol > lactose > maltitol > sorbitol > sodium chloride. Composite granules of the particle size range 355-710 µm were superior in increasing the drug dissolution rate from ordered mixtures. Reducing the carrier particle size decreased the dissolution rate of the drug, as well as the increase in drug concentration. Kinetic modeling of drug release data fitted best the Hixson-Crowell model, which indicates that all the ordered mixture formulations followed the cube root law fairly well.Teško topljivi antidijabetik gliklazid izabran je za proučavanje utjecaja pomoćnih tvari na povećanje oslobađanja. Homogene smjese mikroniziranog gliklazida s laktozom, manitolom, sorbitolom, maltitolom i natrijevim kloridom pripravljene su ručnim tresenjem staklenih bočica s lijekom i pomoćnom tvari/tvarima. Na oslobađanje lijeka utjecali su vrsta vodotopljivog ekscipijensa, dodatak surfaktanta i superdezintegratora, udio lijeka i veličina čestica punila. Sve vodotopljive pomoćne tvari povećavale su oslobađanje ljekovite tvari i to sljedećim redom: manitol > laktoza > maltitol > sorbitol > natrijev klorid. Najbolje oslobađanje lijeka bilo je iz kompozitnih granula veličine 355 do 710 µm. Iz smjesa s manjom veličinom čestica punila i većim udjelom lijeka oslobađanje lijeka bilo je manje. Kinetičko modeliranje oslobađanja najbolje je odgovaralo Hixson-Crowellovom modelu, što ukazuje na to da sve formulacije prilično dobro slijede zakon trećeg korijena

Molecular characterization of induced mutagenesis through gamma radiation using RAPD markers in Jatropha curcas L.

Genetic variability in Jatropha curcas was induced by different doses (5, 10, 15, 20, 25, 30, 35, 40, 45 and 50 kR) of gamma-rays. Gamma radiation induced earliness in flowering and the plants set flowers earlier than that of control, which took longer duration of 327 days for flowering. The improved reproductive and yield parameters such as days taken to first flowering, flowering population, male to female ratio and seed yield per plant were recorded in 25 kR dose and seed germination in 5 and 10 kR treated seeds. Molecular characterization of induced mutants (M1 generation) with 47 Random amplified polymorphic DNA (RAPD) primers showed 65.27% polymorphism. The variability created by gamma rays ranged from 9 to 28%. The 50 kR mutant was found to be the most diverse from control followed by 25 kR mutant. Thus, this integrated approach can be used for carrying out the mutation-assisted breeding and subsequent selection of desired mutants using molecular markers in J. curcas.Keywords: Jatropha curcas, random amplified polymorphic DNA (RAPD), gamma-rays, induced mutagenesis

Gas Sensing and Power Harvesting Polyvinylidene Fluoride Nanocomposites Containing Hybrid Nanotubes

Gas sensing properties at room temperature and energy harvesting performances are realized for the polyvinylidene fluoride (PVDF) nanocomposites containing titanium dioxide (TiO2) nanotubes grown in the presence of carbon nanotubes (CNT). While hydrothermal reaction is practiced for the development of TiO2/CNT hybrid nanotubes, spin coating is done for the nanocomposite films to be deposited on sensing electrodes. Influence of various filler concentrations and the synergistic combination of fillers on the sensing characteristics are studied by recording the response times and the stability of the results. Upon exposure to liquefied petroleum gas, the PVDF/TiO2-CNT (2.5 wt.%) gas sensor shows a sensing response of 0.45 s (400 ppm LPG), approximately nine times higher than the composite containing 2.5 wt.% of TiO2 or 2.5 wt.% CNT. The piezoelectric response of the samples is also recorded and correlated with the synergistic influence of the filler materials. The current study can stimulate a good trend in fabricating self-powered gas sensors from PVDF nanocomposites.Open Access funding provided by the Qatar National Library. This publication was made possible by NPRP Grant 6-282-2-119 from the Qatar National Research Fund (a member of Qatar Foundation). The statements made herein are solely the responsibility of the authors.Scopu

Effect of dietary supplementation of chromium on growth and biochemical parameters of Labeo rohita (Hamilton) fingerlings

A 60-day feeding experiment was conducted to evaluate the effect of dietary chromium on growth, feed efficiency and biochemical parameters of Labeo rohita fingerlings. Four isonitogenous (crude protein 35%) and isocaloric (415 k cal 100 g-1) experimental feeds were prepared by supplementing different levels of dietary chromium picolinate viz., control (0.0 mg kg-1), T1 (0.4 mg kg-1), T2 (0.8 mg kg-1) and T3 (1.2 mg kg-1). Weight gain WG (%), specific growth rate (SGR), feed efficiency ratio (FER) and protein efficiency ratio (PER) and apparent net protein utilisation (ANPU %) were significantly improved (p<0.05) when chromium was supplemented at 0.8 mg kg-1 feed. The protein retention (PR %) value increased with the dose of chromium, showing the highest value in T2 group. Chromium supplementation significantly increased (p<0.05) liver glycogen in T1 and T2 groups but decrease was observed at high level of chromium supplementation in T3 group. Chromium supplementation significantly reduced (p<0.05) serum cholesterol and triglycerides in all the experimental groups compared to control showing the highest reduction in T2 group. The serum high density lipoproteins-cholesterol (HDL-C) was increased (p<0.05)in all experimental groups due to chromium supplementation and the highest blood HDL-C was observed in T2 group. However, no difference (p<0.05) in the serum low density lipoproteins-cholesterol (LDL-C) and phospholipid was observed in any of the experimental groups. Similarly, highest muscle protein as well as lowest liver AST and ALT were observed in T2 group. The results of the present study indicates that growth, feed utilisation and biochemical parameters in Labeo rohita can be significantly improved by feeding the fingerlings with chromium picolinate supplemented diet (0.8 mg kg-1feed)

Diversity and Recognition Efficiency of T Cell Responses to Cancer

BACKGROUND: Melanoma patients vaccinated with tumor-associated antigens frequently develop measurable peptide-specific CD8+ T cell responses; however, such responses often do not confer clinical benefit. Understanding why vaccine-elicited responses are beneficial in some patients but not in others will be important to improve targeted cancer immunotherapies. METHODS AND FINDINGS: We analyzed peptide-specific CD8+ T cell responses in detail, by generating and characterizing over 200 cytotoxic T lymphocyte clones derived from T cell responses to heteroclitic peptide vaccination, and compared these responses to endogenous anti-tumor T cell responses elicited naturally (a heteroclitic peptide is a modification of a native peptide sequence involving substitution of an amino acid at an anchor residue to enhance the immunogenicity of the peptide). We found that vaccine-elicited T cells are diverse in T cell receptor variable chain beta expression and exhibit a different recognition profile for heteroclitic versus native peptide. In particular, vaccine-elicited T cells respond to native peptide with predominantly low recognition efficiency—a measure of the sensitivity of a T cell to different cognate peptide concentrations for stimulation—and, as a result, are inefficient in tumor lysis. In contrast, endogenous tumor-associated-antigen-specific T cells show a predominantly high recognition efficiency for native peptide and efficiently lyse tumor targets. CONCLUSIONS: These results suggest that factors that shape the peptide-specific T cell repertoire after vaccination may be different from those that affect the endogenous response. Furthermore, our findings suggest that current heteroclitic peptide vaccination protocols drive expansion of peptide-specific T cells with a diverse range of recognition efficiencies, a significant proportion of which are unable to respond to melanoma cells. Therefore, it is critical that the recognition efficiency of vaccine-elicited T cells be measured, with the goal of advancing those modalities that elicit T cells with the greatest potential of tumor reactivity

Fog computing security: a review of current applications and security solutions

Fog computing is a new paradigm that extends the Cloud platform model by providing computing resources on the edges of a network. It can be described as a cloud-like platform having similar data, computation, storage and application services, but is fundamentally different in that it is decentralized. In addition, Fog systems are capable of processing large amounts of data locally, operate on-premise, are fully portable, and can be installed on heterogeneous hardware. These features make the Fog platform highly suitable for time and location-sensitive applications. For example, Internet of Things (IoT) devices are required to quickly process a large amount of data. This wide range of functionality driven applications intensifies many security issues regarding data, virtualization, segregation, network, malware and monitoring. This paper surveys existing literature on Fog computing applications to identify common security gaps. Similar technologies like Edge computing, Cloudlets and Micro-data centres have also been included to provide a holistic review process. The majority of Fog applications are motivated by the desire for functionality and end-user requirements, while the security aspects are often ignored or considered as an afterthought. This paper also determines the impact of those security issues and possible solutions, providing future security-relevant directions to those responsible for designing, developing, and maintaining Fog systems