Soil Baiting, Rapid PCR Assay and Quantitative Real TIME PCR to Diagnose Late Blight of Potato in Quarantine Programs

Phytophthora infestans (mont) de Bary is a pathogen of great concern across the globe, and accurate detection is an important component in responding to the outbreaks of potential disease. Although the molecular diagnostic protocol used in regulatory programs has been evaluated but till date methods implying direct comparison has rarely used. In this study, a known area soil samples from potato fields where light blight appear every year (both A1 and A2 mating type) was assayed by soil bait method, PCR assay detection and quantification of the inoculums. Suspected disease symptoms appeared on bait tubers were further confirmed by rapid PCR, inoculums were quantified through Real Time PCR, which confirms presence of P. infestans. These diagnostic methods can be highly correlated with one another. Potato tuber baiting increased the sensitivity of the assay compared with direct extraction of DNA from tuber and soil samples. Our study determines diagnostic sensitivity and specificity of the assays to determine the performance of each method. Overall, molecular techniques based on different types of PCR amplification and Real-time PCR can lead to high throughput, faster and more accurate detection method which can be used in quarantine programmes in potato industry and diagnostic laboratory

Genomic signatures of the interactions between the human innate immune system and retroviruses

The human genome is under constant invasion by internal and external pathogens. To control the activity of these pathogens, the host is equipped with innate and adaptive immune mechanisms. The innate immunity, which is comprised of several mechanisms, aids the adaptive immune system and is the first line of defence against invasive pathogens. An important innate immunity mechanism against viral infection is the restriction factors. These intracellular components recognise viral genomic contents inside the host cells and either directly induce lethal mutations or indirectly inhibit the virus by signalling cascades through other immune components. The survival of a virus thus depends on its ability to evolve and avoid genomic patterns/structures that are identifiable by the host immune system. Consequently, the host-induced evolution is expected to play a role in shaping viral genomes. The aim of this thesis is to understand this complex interaction between the host antiretroviral factors and the exogenous (e.g. HIV) as well as the endogenous (e.g. HERV-K) retroviruses by investigating genomic signatures on the viral sequences. To achieve this, novel computational methods utilising motif representation were developed as part of this thesis, and a large number of HIV and human retroelement sequences were obtained and analysed using these methods. Results of these analyses suggest that 1) several CG motifs are severely depleted in the HIV genome, which can be due to recognition of immune stimulatory genomic patterns by pattern recognition receptors such as Toll-like receptors; 2) the genomes of hypermutated HIV and some families of retroelements such as HERV-K contain footprints of G-to-A mutation by the human editing enzymes APOBEC3 but 3) such a footprint is missing in the normal (non-hypermutated) HIV population. Furthermore, the thesis also presents a novel way to identify hypermutated signatures in retroviral genomes, devised by APOBEC3 enzymes without the need for a reference genome. Overall, the findings of this thesis sheds light on the important role of the host defence mechanisms in the genomic evolution of retroviral genomes and would aid in understanding the molecular mechanisms of host/virus interactions, which has implications in vaccine development

Quantitative Analysis of Total Phenolic, Flavonoid Contents and HPTLC Fingerprinting for Standardization of Glycyrrhiza glabra Linn. Roots

The Glycyrrhiza glabra Linn is an important medicinal plant and its roots are commonly used in various indigenous system of medicine to cure many acute and chronic diseases. The current study was designed to quantify total phenolic, flavonoid contents in G. glabra Linn roots to evaluate the antioxidant potential and to carry out the pharmacognostical investigations along with HPTLC fingerprinting in order to develop the quality control parameters for the standardization of this important medicinal plant. A variety of pharmacognostical investigations e.g., extractive values, total ash, water soluble ash, and acid insoluble ash, moisture content, loss on drying, pH and phytochemical screening of G. glabra Linn roots were analyzed as per the standard methods. The content of total phenolics, flavonoids, pesticide residues, aflatoxin and heavy metals were also determined as per the reported methods. The HPTLC fingerprinting of methanolic extract of the G. glabra roots was also carried out using CAMAG-HPTLC system. The results of phytochemicals investigations revealed the presence of carbohydrates, phenolic compounds, flavonoids, alkaloids, proteins, saponins, lipids, sterols and tannins in various solvent extracts. Total phenolic and flavonoid contents in methanolic extract were found to be 7.47 mg/gm and 2.25 µg/gm, respectively. Heavy metals concentrations were found to be under the standard limits. Aflatoxins and pesticides residues were not detected in the extract. The standard parameters established through this study may prove important tools for authentication, identification, purification and standardization of G. glabra Linn roots in herbal pharmaceutical industries

Production and Maintenance in Industries: Impact of Industry 4.0

Purpose: Production industries are undergoing a digital transition, referred to as the fourth industrial revolution or Industry 4.0, as a result of rapidly expanding advances in information and communication technology. The purpose of this research is to provide a conceptual insight into the impact of unique capabilities from the fourth industrial revolution on production and maintenance tasks in terms of providing the existing production companies a boost by making recommendations on areas and tasks of great potential. Design/methodology/approach: A survey and a literature review are among the research methods used in the research. The survey collected empirical data using a semi-structured questionnaire, which provided a broad overview of the company's present condition in terms of production and maintenance, resulting in more comprehensive and specific information regarding the study topics. Findings: The study points out that, the implementation of I4.0-technology leads to an increase in production, asset utilization, quality, reduced machine down time in industries, and maintenance. Sensor technology, big data analysis, cloud technologies, mobile end devices, and real-time location systems are now being implemented to improve production processes and boost organizational competitiveness. Moreover, the study highlights that data acquired throughout the production process is utilized for quality control, predictive maintenance, and automatic production control. Furthermore, I4.0 solutions help companies to be more efficient with assets at each stage of the process, allowing them to have a stronger control on inventories and operational-optimization potential. Originality: The findings of the study was supported by empirical data collected through survey that provides an intangible understanding of the importance of distinctive capabilities from the I4.0 revolution on production and maintenance tasks. In this study, some recommendations and guidelines to enhance these tasks are provided that are vital for existing production companie

Paederia foetida Linn. leaf extract: an antihyperlipidemic, antihyperglycaemic and antioxidant activity

BACKGROUND: The primary objective of the present investigation is to evaluate the antidiabetic, antihyperlidemic and antioxidant activity of the methanolic extract of the Paederia foetida Linn. (PF) leaf extract in the streptozotocin induced diabetic rats. METHODS: Single intraperitoneal injection (IP) of streptozotocin (60 mg/kg body weight) was used for induction of diabetes is swiss albino (wistar strain) rats. The induction of diabetes was confirmed after 3 days as noticing the increase in blood sugar level of tested rats. PF at a once a daily dose of 100 mg/kg, 250 mg/kg, 500 mg/kg, p.o. along with glibenclamide 10 mg/kg, p.o. was also given for 28 days. On the 28(th) day rats from all the groups fasted overnight fasted and the blood was collected from the puncturing the retro orbit of the eye under mild anesthetic condition. There collected blood sample was used to determine the antihyperlipidemic, hypoglycemic and antioxidant parameters. RESULTS: The oral acute toxicity studies did not show any toxic effect till the dose at 2000 mg/kg. While oral glucose tolerance test showed better glucose tolerance in tested rats. The statistical data indicated that the different dose of the PF significantly increased the body weight, hexokinase, plasma insulin, high density lipoprotein cholesterol, superoxide dismutase, catalase and glutathione peroxides. It also decreases the level of fasting blood glucose, total cholesterol, triglycerides, low density lipoprotein cholesterol, very low density lipoprotein cholesterol, malonaldehyde, glucose-6-phosphate, fructose-1-6-biphosphate and glycated hemoglobin in STZ induced diabetic rats. The histopathology of STZ induce diabetic rats, as expected the test dose of PF extract considerably modulates the pathological condition of various vital organ viz. heart, kidney, liver, pancreas as shown in the histopathology examinations. CONCLUSIONS: Our investigation has clearly indicated that the leaf extract of Paederia foetida Linn. showed remarkable antihyperglycemic activity due to its possible systematic effect involving in the pancreatic and extra pancreatic mechanism. Forever, the antihyperlipidemic activity was exerted possible by lowering the higher level of lipid profile and decreasing the intercalated disc space in the heart. The antioxidant activity of extract was due to inhibition of lipid peroxidation and increasing the SOD, GPx and CAT. It was corroborate that the extract shown the Paederia foetida Linn leaves potential to be act as antidiabetic, antihyperlipidemic and antioxidant properties

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival