NOPR Collection:

RNA interference in agricultural insect pest management: Status and perspectives

Fri, 01 Nov 2024 00:00:00 GMT

Title: RNA interference in agricultural insect pest management: Status and perspectives Authors: Prakash, Nitish Ranjan; Ahuja, Amit; Kumar, Santosh; Saini, Varun; Anokhe, Archana; Jayaswal, Deepanshu; Kumar, Kuldeep Abstract: In recent years, RNA interference (RNAi) has emerged as a potential tool to manage the pest population in agricultural fields and storage systems, however so far it has not come up in a big way. RNAi is a basic molecular biology mechanism occurring in all eukaryotic cells wherein a dsRNA (specific to a particular target messenger RNA) is used to inhibit/silence/knockdown gene expression. This dsRNA undergoes nucleolytic cleavage and several modifications to bind the complementary mRNA and induce endo-nucleolytic cleavage or translation repression. This basic molecular biological phenomenon can be exploited to silence the key genes (virulence and disease progression, toxin production, pest-specific key genes) in pests thus challenging their fecundity and survival. This mechanism is suitable for managing most insects (lepidopteran, dipteran, coleopteran, hemipteran, etc.) and nematodes. Application of RNAi differs based on the mode of delivery system of dsRNA to the pest. The efficiency and efficacy of RNAi depend upon targeted genes, type of dsRNA produced, length of dsRNA, feeding methods, stability of dsRNA in insect guts, and easy uptake by gut epithelium, etc. However, this technology is not utilised up to its full potential and therefore the present review is aimed to assess the possibility of RNAi in agricultural pest control and the way forward. Page(s): 851-861

Effect of chemical mutagens on expression of therapeutic protein-streptokinase in wild strain Streptococcus equinus VIT_VB2

Fri, 01 Nov 2024 00:00:00 GMT

Title: Effect of chemical mutagens on expression of therapeutic protein-streptokinase in wild strain Streptococcus equinus VIT_VB2 Authors: Babu, Vaishnavi; V, Mohanasrinivasan; C, George Priya Doss; Ganesh, Sanjeev K; C, Subathra Devi Abstract: Streptokinase breaks down the clot in myocardial infarction, affecting three million people globally. The current study, enhanced the production of industrially important fibrinolytic enzyme, streptokinase (SK), this can be used to reduce death rate due to myocardial infarction. The ultra-violet (UV) mutated strain, UVSE6 of S. equinus VIT_VB2 showed maximum substrate specific-SK activity (864±0.6 IU mL-1) and partial clot lysis (79%). Hence, the mutant strain UVSE6 was further enhanced by chemical mutagenesis. The improved mutant strain EMS1 after chemical mutagenesis showed maximum SK activity (1004.5±0.7 IU mL-1) and partial clot lysis activity (89%), significantly higher than wild strain. The amidolytic activity of purified SK from mutant strain EMS1 of S. equinus VIT_VB2 was found to be 8253 ± 1.6 IU. The molecular weight of SK was determined as 47 kDa by SDS-PAGE and purity of SK was confirmed by HPLC (retention time: 2.82 min). Presence of SK gene isolated from EMS1 mutant strain S. equinus VIT_VB2 was confirmed using molecular gene sequencing (1200 bp). Structural analysis reveals 3.7% of the amino acid residues in outlier region in the wild type model increase in the mutant. The variation of amino acid in the sequences is observed in RAMPAGE analysis. Page(s): 862-872

Probing the risk markers of mild cognitive impairment and Alzheimer's disease in alloxan-induced diabetic rats

Fri, 01 Nov 2024 00:00:00 GMT

Title: Probing the risk markers of mild cognitive impairment and Alzheimer's disease in alloxan-induced diabetic rats Authors: Varadaiah, Yogaraje Gowda C; Sivanesan, Senthilkumar; Howell, Mathew D; Rajadas, Jayakumar; Kavuri, Suchitra; Vijayaraghavan, Rajagopalan; RT, Kashinath Abstract: Alzheimer's disease (AD) exemplifies a form of diabetes mellitus that selectively affects the brain. In this study we evaluated serum markers and genes linked to AD and investigated the possible link between AD and diabetes. We measured the serum levels of insulin, lactate, citrulline, argininosuccinate, homocitrulline, and AD-associated lipids (2,4-dihydroxybutanoic acid and sphinganine-1-phosphate). We used semi-quantitative reverse transcription polymerase chain reaction to determine the relative gene expression of Rest, Bace1, and Rtn3 in the brain. Western blotting was used to determine the relative protein expression of REST, BACE1, and RTN3 in the brain. Serum from diabetic rats showed significantly higher levels of citrulline, homocitrulline, lactate, 2,4-dihydroxybutanoic acid, and sphinganine-1-phosphate, but lower levels of argininosuccinate and insulin compared with serum from control rats. At the gene level, Bace1 and Rest were significantly increased, whereas Rtn3 was considerably reduced in diabetic compared with control rat brains. At the protein level, BACE1 and RTN3 were significantly increased whereas REST was significantly reduced in diabetic compared with control rat brains. These findings indicate that hyperglycemia and insulin resistance in the brain may be the common element that connects urate turnover and alterations in neural regulatory genes. Page(s): 873-883

Exploring urinary biomarkers for the diagnosis of diabetic and hypertensive chronic kidney disease: A promising pilot study

Fri, 01 Nov 2024 00:00:00 GMT

Title: Exploring urinary biomarkers for the diagnosis of diabetic and hypertensive chronic kidney disease: A promising pilot study Authors: Saseevan, Sumana; Nishanthi, Nadeesha; Rajapakse, Sanath; Magana-Arachchi, Dhammika Abstract: In the current clinical setting, conventional serum biomarkers such as serum creatinine (Scr) and estimated glomerular filtration rate (eGFR) have several lapses in chronic kidney disease (CKD) diagnosis. Diagnosing CKD using non-invasive methods is crucial for implementing prompt therapeutic interventions and preventing disease progression. This study aims to identify the potential diagnostic urinary biomarkers and their correlation with existing renal markers, Scr, eGFR, and proteinuria in diabetic and hypertensive CKD. RNA was extracted from eighty-two urine samples of CKD patients and healthy controls (HC) and reverse transcribed for gene expression analysis using quantitative polymerase chain reactions. The expression of NGAL, MMP9, ANXA3, OLFM4, PI3, and PRMT3 genes was analyzed relative to the reference gene, B2M. Fold changes (FC) in gene expression in diabetic nephropathy (DN), and hypertensive nephropathy (HT) were calculated against HC. Log2 normalized FC was used to determine significance levels and correlation with existing serum markers. NGAL, ANXA3, and OLFM4 exhibited the highest upregulations in DN with mean Log2FC 1.42, 2.66, and 5.87, respectively. A two-fold increase in NGAL FC was observed in early DN than in late DN, suggesting its potential as an early urinary biomarker for DN. PI3 and MMP9 were upregulated in HT patients with higher FC values. PRMT3 showed a significant negative correlation (P<0.05) in HT patients with Scr (r=−0.738) and proteinuria (r=−0.906). The gene panels including ANXA3, OLFM4, and NGAL, and PI3, PRMT3, and MMP9, could have potential diagnostic value in DN and HT, respectively. Page(s): 884-894