34 research outputs found
Microarray analysis of ncRNA expression patterns in Caenorhabditis elegans after RNAi against snoRNA associated proteins
Different Pretreatment Methods of Lignocellulosic Biomass for Use in Biofuel Production
Lignocellulosic biomasses are carbon neutral and abundantly available renewable bioresource material available on earth. However, the main problem that hinders its frequent use is the tight bonding within its constituents that include cellulose, hemicellulose, and lignin. The selection of pretreatment process depends exclusively on the application. Various pretreatment processes are primarily developed and utilized in effective separation of these interlinked components to take maximum benefit from the constitutes of the lignocellulosic biomasses especially for the production of biofuel. The major pretreatment methods include physical, chemical, thermophysical, thermochemical, and biological approaches. Various aspects of these different pretreatment approaches are discussed in this chapter
Systematic identification of non-coding RNA 2,2,7-trimethylguanosine cap structures in Caenorhabditis elegans
Rearing the Cotton Bollworm, Helicoverpa armigera, on a Tapioca-Based Artificial Diet
The impact of a tapioca-based artificial diet on the developmental rate, life history parameters, and fertility was examined over five consecutive generations for the cotton bollworm, Helicoverpa armigera Hubner (Lepidoptera: Noctuidae), a highly polyphagous pest of many agricultural crops. The study showed that when fed the tapioca-based artificial diet during larval stage, larval and pupal developmental period, percent pupating, pupal weight, emergence rate of male and female, longevity, fecundity and hatching were non-significantly different than that of the control agar-based artificial diet. Moreover, the cost to rear on tapioca-based diet approached 2.13 times less than the cost of rearing on the agar-based artificial diet. These results demonstrate the effectiveness and potential cost savings of the tapioca-based artificial diet for rearing H. armigera
Biosynthesis and characterization of silver nanoparticles from Cedrela toona leaf extracts: An exploration into their antibacterial, anticancer, and antioxidant potential
This research work aims to synthesize environmentally benign and cost-effective metal nanoparticles. In this current research scenario, the leaf extract of Cedrela toona was used as a reducing agent to biosynthesize silver nanoparticles (AgNPs). The synthesis of AgNPs was confirmed by the color shift of the reaction mixture, i.e., silver nitrate and plant extract, from yellow to dark brown colloidal suspension and was established by UV-visible analysis showing a surface plasmon resonance band at 434 nm. Different experimental factors were optimized for the formation and stability of AgNPs, and the optimum conditions were found to be 1 mM AgNO3 concentration, a 1:9 ratio of extract/precursor, and an incubation temperature of 70°C for 4 h. The Fourier transform infrared spectroscopy spectra indicated the presence of phytochemicals in the leaf extract that played the role of bioreducing agents in forming AgNPs. X-ray diffraction patterns confirmed the presence of AgNPs with a mean size of 25.9 nm. The size distribution and morphology of AgNPs were investigated by scanning electron microscopy, which clearly highlighted spherical nanoparticles with a size distribution of 22–30 nm with a mean average size of 25.5 nm. Moreover, prominent antibacterial activity was found against Enterococcus faecalis (21 ± 0.5 mm), Bacillus subtilis (20 ± 0.9 mm), Pseudomonas aeruginosa (18 ± 0.3 mm), Staphylococcus aureus (16 ± 0.7 mm), Klebsiella pneumoniae (16 ± 0.3 mm), and Escherichia coli (14 ± 0.7 mm). In addition, antioxidant activity was determined by DPPH and ABTS assays. Higher antioxidant activity was reported in AgNPs compared to the plant extract in both DPPH (IC50 = 69.62 µg·ml−1) and ABTS assays (IC50 = 47.90 µg·ml−1). Furthermore, cytotoxic activity was also investigated by the MTT assay against MCF-7 cells, and IC50 was found to be 32.55 ± 0.05 µg·ml−1. The crux of this research is that AgNPs synthesized from the Cedrela toona leaf extract could be employed as antibacterial, antioxidant, and anticancer agents for the treatment of bacterial, free radical-oriented, and cancerous diseases
Profiling Caenorhabditis elegans non-coding RNA expression with a combined microarray
Small non-coding RNAs (ncRNAs) are encoded by genes that function at the RNA level, and several hundred ncRNAs have been identified in various organisms. Here we describe an analysis of the small non-coding transcriptome of Caenorhabditis elegans, microRNAs excepted. As a substantial fraction of the ncRNAs is located in introns of protein-coding genes in C.elegans, we also analysed the relationship between ncRNA and host gene expression. To this end, we designed a combined microarray, which included probes against ncRNA as well as host gene mRNA transcripts. The microarray revealed pronounced differences in expression profiles, even among ncRNAs with housekeeping functions (e.g. snRNAs and snoRNAs), indicating distinct developmental regulation and stage-specific functions of a number of novel transcripts. Analysis of ncRNA–host mRNA relations showed that the expression of intronic ncRNA loci with conserved upstream motifs was not correlated to (and much higher than) expression levels of their host genes. Even promoter-less intronic ncRNA loci, though showing a clear correlation to host gene expression, appeared to have a surprising amount of ‘expressional freedom’, depending on host gene function. Taken together, our microarray analysis presents a more complete and detailed picture of a non-coding transcriptome than hitherto has been presented for any other multicellular organism
The functional characterization of long noncoding RNA SPRY4-IT1 in human melanoma cells
Expression of the long noncoding RNA (lncRNA) SPRY4-IT1 is low in normal human melanocytes but high in melanoma cells. siRNA knockdown of SPRY4-IT1 blocks melanoma cell invasion and proliferation, and increases apoptosis. To investigate its function further, we affinity purified SPRY4-IT1 from melanoma cells and used mass spectrometry to identify the protein lipin 2, an enzyme that converts phosphatidate to diacylglycerol (DAG), as a major binding partner. SPRY4-IT1 knockdown increases the accumulation of lipin2 protein and upregulate the expression of diacylglycerol O-acyltransferase 2 (DGAT2) an enzyme involved in the conversion of DAG to triacylglycerol (TAG). When SPRY4-IT1 knockdown and control melanoma cells were subjected to shotgun lipidomics, an MS-based assay that permits the quantification of changes in the cellular lipid profile, we found that SPRY4-IT1 knockdown induced significant changes in a number of lipid species, including increased acyl carnitine, fatty acyl chains, and triacylglycerol (TAG). Together, these results suggest the possibility that SPRY4-IT1 knockdown may induce apoptosis via lipin 2-mediated alterations in lipid metabolism leading to cellular lipotoxicity
Convalescent plasma in patients admitted to hospital with COVID-19 (RECOVERY): a randomised controlled, open-label, platform trial
SummaryBackground Azithromycin has been proposed as a treatment for COVID-19 on the basis of its immunomodulatoryactions. We aimed to evaluate the safety and efficacy of azithromycin in patients admitted to hospital with COVID-19.Methods In this randomised, controlled, open-label, adaptive platform trial (Randomised Evaluation of COVID-19Therapy [RECOVERY]), several possible treatments were compared with usual care in patients admitted to hospitalwith COVID-19 in the UK. The trial is underway at 176 hospitals in the UK. Eligible and consenting patients wererandomly allocated to either usual standard of care alone or usual standard of care plus azithromycin 500 mg once perday by mouth or intravenously for 10 days or until discharge (or allocation to one of the other RECOVERY treatmentgroups). Patients were assigned via web-based simple (unstratified) randomisation with allocation concealment andwere twice as likely to be randomly assigned to usual care than to any of the active treatment groups. Participants andlocal study staff were not masked to the allocated treatment, but all others involved in the trial were masked to theoutcome data during the trial. The primary outcome was 28-day all-cause mortality, assessed in the intention-to-treatpopulation. The trial is registered with ISRCTN, 50189673, and ClinicalTrials.gov, NCT04381936.Findings Between April 7 and Nov 27, 2020, of 16 442 patients enrolled in the RECOVERY trial, 9433 (57%) wereeligible and 7763 were included in the assessment of azithromycin. The mean age of these study participants was65·3 years (SD 15·7) and approximately a third were women (2944 [38%] of 7763). 2582 patients were randomlyallocated to receive azithromycin and 5181 patients were randomly allocated to usual care alone. Overall,561 (22%) patients allocated to azithromycin and 1162 (22%) patients allocated to usual care died within 28 days(rate ratio 0·97, 95% CI 0·87–1·07; p=0·50). No significant difference was seen in duration of hospital stay (median10 days [IQR 5 to >28] vs 11 days [5 to >28]) or the proportion of patients discharged from hospital alive within 28 days(rate ratio 1·04, 95% CI 0·98–1·10; p=0·19). Among those not on invasive mechanical ventilation at baseline, nosignificant difference was seen in the proportion meeting the composite endpoint of invasive mechanical ventilationor death (risk ratio 0·95, 95% CI 0·87–1·03; p=0·24).Interpretation In patients admitted to hospital with COVID-19, azithromycin did not improve survival or otherprespecified clinical outcomes. Azithromycin use in patients admitted to hospital with COVID-19 should be restrictedto patients in whom there is a clear antimicrobial indication
Fungal flora and aflatoxin contamination in Pakistani wheat kernels (Triticum aestivum L.) and their attribution in seed germination
This study aimed to isolate fungal pathogens and to subsequently quantify aflatoxin (AF; B1 + B2 + G1 + G2) contamination in wheat crops grown in Pakistan. Accordingly, a total of 185 wheat samples were collected from different areas of Pakistan and numerous potent fungal pathogens were isolated. AF contamination attributed to the presence of intoxicating fungal pathogens and resulting metabolic activities were quantified using a high performance liquid chromatography-fluorescence detector coupled with postcolumn derivatization. Additionally, the effect of fungal pathogens on seed germination was also examined. The results obtained showed that 50% of tested wheat samples were found to be contaminated with a diverse range of fungal species. The rate of recurrence of fungal pathogens were Aspergillus 31%, Penicillium 9%, Fusarium 8%, Rhizopus 3%, and Alternaria 2%. The presence of Tilletia indica and Claviceps purpurea species was found to be inevident in all tested wheat samples. AFB1 contamination was detected in 48 (26.0%) samples and AFB2 in 13 (7.0%) samples. AFG1 and AFG2 were not found in any of the tested samples. The contamination range of AFB1 and AFB2 was 0.05–4.78 μg/kg and 0.02–0.48 μg/kg, respectively. The total amount of AFs (B1 + B2) found in 48 (26.0%) samples had a mean level of 0.53 ± 0.40 μg/kg and a contamination range of 0.02–5.26 μg/kg. The overall results showed that in 137 (74.0%) samples, AFs were not found within detectable limits. Furthermore, in 180 (97.2%) samples, AF levels were found to be below the maximum tolerated levels (MTL) recommended by the European Union (4 μg/kg). In five (2.7%) samples, AF contamination was higher than the MTL of the European Union. However, these samples were fit for human consumption with reference to the MTL (20 μg/kg) assigned by the USA (Food and Drug Administration and Food and Agriculture Organization) and Pakistan (Pakistan Standards and Quality Control Authority). Germination rates in healthy and contaminated wheat kernels were 84.6% and 45.2%, respectively. Based on the obtained results, it was concluded that the levels of fungal pathogen and AF contamination in Pakistani-grown wheat are not a potential threat to consumer health. However, control procedures along with a strict monitoring policy are mandatory to further minimize the prevalence of fungal carriers and the potency of AFs in crops cultivated in Pakistan