31 research outputs found
Regulatory Model of Mirna-125 in Leukemia
MicroRNAs (miRNAs) are small, subtypes of RNA that are 22-25 nucleotides in length that regulate the gene expression by targeting homologous sequences in messenger RNA (mRNA). Their unusual expressions have been observed in various types of cancers. In this paper we first briefly outline of multiple pairwise alignment of primiRNA- 125 of three organisms and secondary structure. We then discuss in detail on binding sites of miRNA-125 with different target genes to known the binding energy using computational tool. Finally, we discuss on mechanism of miRNA-125 over-expression causes different types of leukemia due to abnormal expression of gene present on chromosome 21. Thus we have reported that one of the major causes of leukemia is abnormal expression of miRNA- 125.
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Computational prediction and identification of miRNAs in ssRNA viruses
MiRNAs are conserved endogenous small non-coding RNA mols. play a very important role in posttranscriptional regulation of gene by binding to mRNA.miRNAs have been identified in plants, animals, invertebrates and even in viruses. Recent studies have proved that as like DNA viruses even cytoplasmic RNA viruses encodes miRNA by noncanonical pathway.To find the miRNAs encoded by both (+) ssRNA and (-) ssRNA viruses, we used computational tool VMir to screen the complete genomes of selected RNA viruses. The predicted precursor miRNA hairpin structures obtained from VMir were further analyzed in miPred to identify real and pseudo miRNA. The real miRNAs analyzed in blastx program to exclude protein coding sequences and then finally identified mature miRNAs by using matureBayes.In our studies we analyzed 45 viral genomes, finally we obsd. that more miRNAs encoded by (-) ssRNA than the (+) ssRNA
Computational identification and characterization of putative miRNAs in Nasonia species
MicroRNAs are important at post transcriptional regulation in eukaryotes. Nasonia genus is becoming increasingly popular model in present days due to genetic advantages it possesses over Drosophila. Nasonia species are found distributed throughout the world, expect for N. longicornis, and N. giraulti. In this study, we use the sequential method of blasting all known invertebrate miRNA genes against the Nasonia vitripennis, Nasonia longicornis, and Nasonia giraulti genomes. We identify 40, 31 and 29 putative pre-miRNAs and mature sequences in N. vitripennis, N. giraulti and N. longicornis resp. A cross species comparison of putative miRNA sequences and their statistical characteristics reveals that there are no huge differences between the species, except for few miRNAs which are reported. We also find that the minimal folding energy index for three Nasonia species pre-miRNA's av. is around -0.85 ± 0.11. Further, we report that U is predominant at the 5' end of mature sequence, which being a typical characteristic of plant miRNAs. Using MiRanda, we predict nearly 471 potential sites in the N. vitripennis genome. Thus concluding our study to be the beginning of understanding the Nasonia's non coding RNAs and may play an important role in effective pest management in near future
Carboxylesterases from the seeds of an underutilized legume, Mucuna pruriens; Isolation, purification and characterization
Two carboxylesterases (ME-III and ME-IV) have been purified to apparent homogeneity from the seeds of Mucuna pruriens employing ammonium sulfate fractionation, cation exchange chromatography on CM-cellulose, gel-permeation chromatography on Sephadex G-100 and preparative PAGE. The homogeneity of the purified preparations was confirmed by polyacrylamide gel electrophoresis (PAGE), gel-electrofocussing and SDS-PAGE. The molecular weights determined by gel-permeation chromatography on Sephadex G-200 were 20.89 kDa (ME-III) and 31.62 kDa (ME-IV). The molecular weights determined by SDS-PAGE both in the presence and absence of 2-mercaptoethanol were 21 kDa (ME-III) and 30.2 kDa (ME-IV) respectively, suggesting a monomeric structure for both the enzymes. The enzymes were found to have Stokes radius of 2.4 nm (ME-III) and 2.7 nm (ME-IV). The isoelectric pH values of the enzymes, ME-III and ME-IV, were 6.8 and 7.4, respectively. ME-III and ME-IV were classified as carboxylesterases employing PAGE in conjunction with substrate and inhibitor specificity. The K m of ME-III and ME-IV with 1-naphthyl acetate as substrate was 0.1 and 0.166 mM while with 1-naphthyl propionate as substrate the K m was 0.052 and 0.0454 mM, respectively. As the carbon chain length of the acyl group increased, the affinity of the substrate to the enzyme increased indicating hydrophobic nature of the acyl group binding site. The enzymes exhibited an optimum temperature of 45 °C (ME-III) and 37 °C (ME-IV), an optimum pH of 7.0 (ME-III) and 7.5 (ME-IV) and both the enzymes (ME-III and ME-IV) were stable up to 120 min at 35 °C. Both the enzymes were inhibited by organophosphates (dichlorvos and phosphamidon), but resistant towards carbamates (carbaryl and eserine sulfate) and sulphydryl inhibitors (p-chloromercuricbenzoate, PCMB). © 2011 Elsevier Ltd. All rights reserved
Esterase Activity from the Germinated Jatropha curcas Seeds in Different Extraction buffers.
The buffer solution plays a major role in protein stability and activity, thereby making the selection of a buffer to achievemaximum activity for a protein will be a formidable challenge. The present work constitutes
an extension of this investigation to esterases from germinated Jatrophacurcas seeds. The 0.1 M NaCl solution, 0.1 M phosphate buffer pH7.0, 0.1 M citrate buffer pH 5.0 and 0.1 M Tris-HCl buffer pH 8.5, 0.1 M NaOH and distill water were used to extract protein from germinated Jatrophacurcas
seeds. The esterase activity and specific activity for NaCl solution, phosphatebuffer, citrate buffer, Tris-HCl buffer, NaOH and Distilled water was 9.07, 8.6, 8.2, 6.46, 0.07 and 4.98 μmoles/min/gm
and 0.09258, 0.0905, 0.088, 0.0715 0.0003 and 0.081 IU/mg, respectively.The Native-PAGE analysis showed the esterase
enzyme activity in different extraction buffer.Among 13 esterase bands, 8 esterolytic bands were major bands
(band no 1,3, 6, 7, 8, 11, 12 and 13)and remaining were minor bands.The amount of proteins and esterase activity were found to bethe highest when extracted with 0.1 M NaCl solution
CHARACTERIZATION OF ALPHA-AMYLASE FROM THE SEEDS OF Mucuna pruriens
Amylases are hydrolytic enzymes which are widely distributed in nature, animals, plants and
microorganisms. Amylases are of great significance in present-day biotechnology. In present study,
amylases are isolated from the soaked seeds of Mucuna pruriens under extreme acidic conditions.
Conventional protein purification techniques such as salt fractionation, ion exchange chromatography on
CM-cellulose and sephadex G-75 was employed for the purification of amylase from the seeds of
Mucuna pruriens. The amylase activity was eluted in one peak. The specific activity and yield of the
purified amylase was 6.25 and 29.99, respectively. Native PAGE, SDS-PAGE and gel electrofocussing
were employed to establish homogeneity of the purified amylase. SDS-PAGE and gel-filtration
chromatography on sephadex G-75 was used to determine the molecular weight of the purified amylase.
The purified amylase was nearly homogenous and its molecular weight was found to be 78.4 kDa. The
optimum pH and temperature of the purified amylase were 7.0 and 50oC, respectively. The isolectric pH
of the purified amylase was 7.2 and the activity was linear up to 60 minutes
Variations in the esterase activity during the germination period of Jatropha curcas seeds
Germination brings out the synthesis or activation of enzymes responsible for the degradation of seeds reserves. Among these enzymes, esterases are involved in the metabolic processes of germination and maturation of plants. They are constitutively expressed in seeds during germination to release the reserve materials for the growing embryo. In the present study, total protein content and esterase activity was monitored in germinating Jatropha curcas seeds. The esterase activity and specific activity observed were 9.07 µmoles/min/gm and 0.09258 IU/mg, respectively. Electophoretic analysis for esterase activity showed thirteen bands of esterases, among these 8 esterolytic bands were major and remaining were minor bands. The protein content and esterase activity decreased on 2nd, 4th, 5th and 8th day of seed germination and activity increased on 3rd, 6th, 7th day of germination. Similarly esterase activity increased on 7th day and decreased on 8, 9 and 10th day in the shoot tissue. ÂÂ
Preparation, Characterization and In Vitro Drug Release Studies of 6-mercaptopurine Thin Film
Oral thin films of 6-mercaptopurine were fabricated from mucoadhesive polymer, chitosan and polyvinylpyrrolidone for the purpose of prolonging drug release and improving its bioavailability. All fabricated film formulations prepared were smooth and translucent, with good flexibility. The weight and thickness of all the formulations were found to be uniform. These films were also evaluated for surface pH, folding endurance, swelling percentage (% S) and in vitro disintegration time. Scanning Electron Microscopy (SEM) and Fourier Transform Infrared (FTIR) were used to evaluate the physico-chemical nature of the films. In-vitro drug release have shown enhanced release profiles for thin films compared to pure drug and the release patterns have been found to be pH dependant. The results of the study reveals that fabrication of 6-MP oral thin film by using solvent cast technology is a simple and an efficient method for drug delivery to achieve desired therapeutic compliance.Keywords: 6-mercaptopurine; In Vitro Drug Release; SEM; FTI
Carboxylesterases from the seeds of an underutilized legume, Mucuna pruriens; isolation, purification and characterization
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Global burden of 288 causes of death and life expectancy decomposition in 204 countries and territories and 811 subnational locations, 1990–2021: a systematic analysis for the Global Burden of Disease Study 2021
BACKGROUND Regular, detailed reporting on population health by underlying cause of death is fundamental for public health decision making. Cause-specific estimates of mortality and the subsequent effects on life expectancy worldwide are valuable metrics to gauge progress in reducing mortality rates. These estimates are particularly important following large-scale mortality spikes, such as the COVID-19 pandemic. When systematically analysed, mortality rates and life expectancy allow comparisons of the consequences of causes of death globally and over time, providing a nuanced understanding of the effect of these causes on global populations. METHODS The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2021 cause-of-death analysis estimated mortality and years of life lost (YLLs) from 288 causes of death by age-sex-location-year in 204 countries and territories and 811 subnational locations for each year from 1990 until 2021. The analysis used 56 604 data sources, including data from vital registration and verbal autopsy as well as surveys, censuses, surveillance systems, and cancer registries, among others. As with previous GBD rounds, cause-specific death rates for most causes were estimated using the Cause of Death Ensemble model-a modelling tool developed for GBD to assess the out-of-sample predictive validity of different statistical models and covariate permutations and combine those results to produce cause-specific mortality estimates-with alternative strategies adapted to model causes with insufficient data, substantial changes in reporting over the study period, or unusual epidemiology. YLLs were computed as the product of the number of deaths for each cause-age-sex-location-year and the standard life expectancy at each age. As part of the modelling process, uncertainty intervals (UIs) were generated using the 2·5th and 97·5th percentiles from a 1000-draw distribution for each metric. We decomposed life expectancy by cause of death, location, and year to show cause-specific effects on life expectancy from 1990 to 2021. We also used the coefficient of variation and the fraction of population affected by 90% of deaths to highlight concentrations of mortality. Findings are reported in counts and age-standardised rates. Methodological improvements for cause-of-death estimates in GBD 2021 include the expansion of under-5-years age group to include four new age groups, enhanced methods to account for stochastic variation of sparse data, and the inclusion of COVID-19 and other pandemic-related mortality-which includes excess mortality associated with the pandemic, excluding COVID-19, lower respiratory infections, measles, malaria, and pertussis. For this analysis, 199 new country-years of vital registration cause-of-death data, 5 country-years of surveillance data, 21 country-years of verbal autopsy data, and 94 country-years of other data types were added to those used in previous GBD rounds. FINDINGS The leading causes of age-standardised deaths globally were the same in 2019 as they were in 1990; in descending order, these were, ischaemic heart disease, stroke, chronic obstructive pulmonary disease, and lower respiratory infections. In 2021, however, COVID-19 replaced stroke as the second-leading age-standardised cause of death, with 94·0 deaths (95% UI 89·2-100·0) per 100 000 population. The COVID-19 pandemic shifted the rankings of the leading five causes, lowering stroke to the third-leading and chronic obstructive pulmonary disease to the fourth-leading position. In 2021, the highest age-standardised death rates from COVID-19 occurred in sub-Saharan Africa (271·0 deaths [250·1-290·7] per 100 000 population) and Latin America and the Caribbean (195·4 deaths [182·1-211·4] per 100 000 population). The lowest age-standardised death rates from COVID-19 were in the high-income super-region (48·1 deaths [47·4-48·8] per 100 000 population) and southeast Asia, east Asia, and Oceania (23·2 deaths [16·3-37·2] per 100 000 population). Globally, life expectancy steadily improved between 1990 and 2019 for 18 of the 22 investigated causes. Decomposition of global and regional life expectancy showed the positive effect that reductions in deaths from enteric infections, lower respiratory infections, stroke, and neonatal deaths, among others have contributed to improved survival over the study period. However, a net reduction of 1·6 years occurred in global life expectancy between 2019 and 2021, primarily due to increased death rates from COVID-19 and other pandemic-related mortality. Life expectancy was highly variable between super-regions over the study period, with southeast Asia, east Asia, and Oceania gaining 8·3 years (6·7-9·9) overall, while having the smallest reduction in life expectancy due to COVID-19 (0·4 years). The largest reduction in life expectancy due to COVID-19 occurred in Latin America and the Caribbean (3·6 years). Additionally, 53 of the 288 causes of death were highly concentrated in locations with less than 50% of the global population as of 2021, and these causes of death became progressively more concentrated since 1990, when only 44 causes showed this pattern. The concentration phenomenon is discussed heuristically with respect to enteric and lower respiratory infections, malaria, HIV/AIDS, neonatal disorders, tuberculosis, and measles. INTERPRETATION Long-standing gains in life expectancy and reductions in many of the leading causes of death have been disrupted by the COVID-19 pandemic, the adverse effects of which were spread unevenly among populations. Despite the pandemic, there has been continued progress in combatting several notable causes of death, leading to improved global life expectancy over the study period. Each of the seven GBD super-regions showed an overall improvement from 1990 and 2021, obscuring the negative effect in the years of the pandemic. Additionally, our findings regarding regional variation in causes of death driving increases in life expectancy hold clear policy utility. Analyses of shifting mortality trends reveal that several causes, once widespread globally, are now increasingly concentrated geographically. These changes in mortality concentration, alongside further investigation of changing risks, interventions, and relevant policy, present an important opportunity to deepen our understanding of mortality-reduction strategies. Examining patterns in mortality concentration might reveal areas where successful public health interventions have been implemented. Translating these successes to locations where certain causes of death remain entrenched can inform policies that work to improve life expectancy for people everywhere. FUNDING Bill & Melinda Gates Foundation