Analysis of reporter proteins GUS and DsRed driven under the control of CaMV35S promoter in syncytia induced by beet cyst nematode Heterodera schachtii in Arabidopsis roots

Background: Cyst nematodes induce specialized feeding structures called syncytia in the plant roots. The expression of CaMV promoter in syncytia has remained topic of debate. The objective of this research was to study the activity of CaMV promoter by using reporter proteins like GUSand DsRed under the control of CaMV35S promoter in syncytia induced by H. schachtii in Arabidopsis roots.Methods: pMAA-Red and pPZP3425 plasmids were used to study expression of GUS and DsRedin syncytia.  The plants were grown in 2% Knop medium under sterile conditions in growth chambers at 25°C in long day conditions. GUS activity in syncytia was studied through staining of syncytia using X-gluc solution. Ds-Red fluorescence in syncytia was detected by using an inverse microscope equipped with UV filter.Results: The expression analysis of DsRed protein driven by CaMV promoter demonstrated that this promoter is active in syncytia at all the time points. All the syncytia showed DsRed expression at 5 dpi. At 7 dpi, 10 dpi and 15 dpi over 90%, 80% and 50% of the syncytia showed DsRedfluorescence respectively. There was very high fluorescence in the syncytia as compared to the uninfected root segments due to high expression. CaMV::GUS lines showed GUS expression in 80% of 5dpi syncytia. However, unlike expression of DsRed, the number of GUS stained syncytia decreased quickly to around 50% at 7 dpi and to about 5% in the 15 dpi syncytia.Conclusions: The results conclude that CaMV promoter is more active in younger syncytia as compared to older syncytia but can be used for expression in syncytia. Moreover, DsRed protein could be used as better reporter for evaluation of gene expression in syncytia as compared to GUS

Arabidopsis Argininosuccinate Lyase and Argininosuccinate Synthase are important for resistance against Pseudomonas syringae

Background: Arginine is one of the important amino acids and is involved in a variety of plant responses against biotic and abiotic stresses. Two important genes Argininosuccinate Synthase (AS) and Argininosuccinate Lyase (AL) are associated with the production of arginine from ornithine. Here we report molecular characterization of Arabidopsis thaliana artificial micro RNA (amiRNA) mutants for the genes AL and AS in response to infection with Pseudomonas syringae DC3000.Methods: Quantitative real time PCR (qRT-PCR) and Promoter::GUS plant lines were used for expression analysis. Artificial micro RNAs (amiRNA) mediated gene silencing was used to generate mutant plant lines. Flood inoculation technique was used for infection test essays with Pseudomonas syringae.Results: Expression analysis of A. thaliana plants harboring promoter AS::GUS construct showed strong promoter activity upon P. syringae infection. Quantitative real time PCR (qRT-PCR) analysis showed that AL and AS expression was strongly induced upon infection with P. syringae. Infection essays for P. syringae showed enhanced susceptibility to virulent (Pto) as well as avirulent (∆avrPto/∆avrPtoB) strains of P. syringae. However, mutant plants infiltered with infiltration medium containing 1 mM L-arginine regain their resistance in comparison with wild type (Col-0) plants.Conclusion: Our findings suggest that genes related to arginine metabolism play a key role in plant defenses during P. syringae infection on A. thaliana. This study revealed that proper functioning of arginine related genes is required to deploy defense response against P. syringae. Decrease in the expression of these genes improves conditions for the growth of pathogen.Keywords: Arabidopsis thaliana; arginine; Argininosuccinate Lyase; Argininosuccinate Synthase; Gene silencing; Pseudomonas syringa

Microencapsulation: A taste and odour masking approach for garlic (Allium sativum) powder

The purpose of this study was to prepare a convenient formulation of garlic powder which has enhanced user compliance by suppressing its characteristic odour and masking the taste. Microcapsules of garlic powder were formulated by solvent evaporation method. Results show >80% yield, >70% drug loading efficiency and slow release profile (promising sustained action for more than 2hrs) and a significant reduction in the aromatic smell and taste evaluated through a single blind cross-over study conducted using ten healthy human volunteers

Insulin Therapy- Common Misconceptions among Diabetics

Abstract Background: To find out misconceptions about insulin among diabetics presenting in outpatient department of a tertiary care hospital. Methods: in this cross-sectional observational study, diabetic patients either male or female, more than 15 year of age, irrespective of their diabetes typing were included. Patient less than 15 year of age, having dementia, having other mental or psychiatric illness were not included. The sampling technique was non probability random sampling . The study tool was a predesigned questionnaire, containing close-ended questions. . Results: Out of  250 respondents,50% were male. About 69% responded that insulin commencement means serious stage of diabetes and 68% considered as an expensive modality . According to 32% insulin usage restricts daily life activities. Conclusion: Diabetic patients of this area had many misconceptions about the insulin thera

Formulation and evaluation of fluconazole loaded nanospongies for improved topical drug delivery

Fluconazole (an antifungal drug) loaded nanosponges (NS) were prepared by an emulsion solvent diffusion method using ethyl cellulose as the polymer. Prepared formulations were evaluated for various physicochemical parameters and in-vitro drug release. NS of fluconazole were discrete, free flowing nanosized particles with perforated orange peel-like morphology as shown by SEM analysis. A topical hydrogel formulation based on the drug loaded NS showed a prolonged release profile for the drug. Kinetic modelling on release data showed that the best fitted model was Higuchi model and release mechanism was by Fickian diffusion. FTIR and PXRD results confirmed the absence of any drug polymer interaction and stability of drug in the delivery system

Unlocking the Story: Piloting Interviews to Illuminate the Journey of Indigenous Chicken Micro-farming

Qualitative interviews provide invaluable insights into human experiences, but they can be challenging, especially for novice researchers. Therefore, conducting a preliminary study becomes crucial to refine interview techniques and improve the overall research process. The present study focuses on the importance of a preliminary study in understanding indigenous chicken micro-farming and aims to gain insights into the interview process and associated issues before conducting a full-scale study. This study represents the first attempt, to the best of our knowledge, to pilot interviews before conducting a comprehensive investigation in the context of indigenous chicken micro-farming. It highlights the methods employed in designing and conducting the preliminary study, as well as the valuable lessons learned throughout the process. The interviews were conducted with indigenous chicken micro-farmers as part of the preparation for a dissertation centered on developing an integrated supply chain model for indigenous chicken in Malaysia. Additionally, this study outlines the modifications made based on the preliminary study for the main study, which subsequently led to the implementation of a pilot study. These adaptations include changes to the interview guide, adjustments in vocabulary to enhance participant understanding, rephrasing of certain questions, and the addition of more interview questions to gain a comprehensive understanding of the issues related to indigenous chicken operations. This study underscores the significance of a preliminary study and its role in informing and shaping the subsequent main study. The findings and insights derived from this preliminary investigation contribute to the development of a robust research design for the main study in indigenous chicken micro-farming. Keywords: indigenous chicken, preliminary study, pilot stud

FIPA-based reference architecture for efficient discovery and selection of appropriate cloud service using cloud ontology

[EN] Cloud computing is considered the latest emerging computing paradigm and has brought revolutionary changes in computing technology. With the advancement in this field, the number of cloud users and service providers is increasing continuously with more diversified services. Consequently, the selection of appropriate cloud service has become a difficult task for a new cloud customer. In case of inappropriate selection of a cloud services, a cloud customer may face the vendor locked-in issue and data portability and interoperability problems. These are the major obstacles in the adoption of cloud services. To avoid these complexities, a cloud customer needs to select an appropriate cloud service at the initial stage of the migration to the cloud. Many researches have been proposed to overcome the issues, but problems still exist in intercommunication standards among clouds and vendor locked-in issues. This research proposed an IEEE multiagent Foundation for Intelligent Physical Agent (FIPA) compliance multiagent reference architecture for cloud discovery and selection using cloud ontology. The proposed approach will mitigate the prevailing vendor locked-in issue and also alleviate the portability and interoperability problems in cloud computing. To evaluate the proposed reference architecture and compare it with the state-of-the-art approaches, several experiments have been performed by utilizing the commonly used performance measures. Analysis indicates that the proposed approach enables significant improvements in cloud service discovery and selection in terms of search efficiency, execution, and response timeAbbas, G.; Mehmood, A.; Lloret, J.; Raza, MS.; Ibrahim, M. (2020). FIPA-based reference architecture for efficient discovery and selection of appropriate cloud service using cloud ontology. International Journal of Communication Systems. 33(14):1-14. https://doi.org/10.1002/dac.4504114331

Synthesis and evaluation of pH dependent hydrogels for controlled release of Venlafaxine HCl

pH dependent hydrogel formulations of venlafexine HCl were prepared by free radical polymerization method using polyethylene glycol as polymer. Various samples were prepared with varying concentration of polymer, monomer and cross-linker to study their effect on gel swelling, diffusion characteristics and drug release. Swelling was found to be increased with increase in pH. Increase in acrylic acid concentration increases swelling while increase in cross-linker concentration has an opposite effect on swelling. Drug release study was performed in pH 1.2, 5.5 and 7.5 for 12 hours at 37 oC and drug release was found to increase in higher pH. Prepared hydrogel preparations were also characterized by PXRD, TGA, SEM and FTIR