Multicistronic lentiviral vectors containing the FMDV 2A cleavage factor demonstrate robust expression of encoded genes at limiting MOI

BACKGROUND: A number of gene therapy applications would benefit from vectors capable of expressing multiple genes. In this study we explored the feasibility and efficiency of expressing two or three transgenes in HIV-1 based lentiviral vector. Bicistronic and tricistronic self-inactivating lentiviral vectors were constructed employing the internal ribosomal entry site (IRES) sequence of encephalomyocarditis virus (EMCV) and/or foot-and-mouth disease virus (FMDV) cleavage factor 2A. We employed enhanced green fluorescent protein (eGFP), O(6)-methylguanine-DNA-methyltransferase (MGMT), and homeobox transcription factor HOXB4 as model genes and their expression was detected by appropriate methods including fluorescence microscopy, flow cytometry, immunocytochemistry, biochemical assay, and western blotting. RESULTS: All the multigene vectors produced high titer virus and were able to simultaneously express two or three transgenes in transduced cells. However, the level of expression of individual transgenes varied depending on: the transgene itself; its position within the construct; the total number of transgenes expressed; the strategy used for multigene expression and the average copy number of pro-viral insertions. Notably, at limiting MOI, the expression of eGFP in a bicistronic vector based on 2A was ~4 times greater than that of an IRES based vector. CONCLUSION: The small and efficient 2A sequence can be used alone or in combination with an IRES for the construction of multicistronic lentiviral vectors which can express encoded transgenes at functionally relevant levels in cells containing an average of one pro-viral insert

Impact of different levels of iron on mitigation of iron chlorosis in varagu CO 3 (Paspalum scrobiculatum. L)

Iron (Fe) deficiency is a major nutritional disorder in crops growing in calcareous soils. Varagu crop are more susceptible to (Fe) deficiency in the early stage of growth and the deficiency is exhibited as chlorosis developing interveinally in the new leaves.  The objective of the present study was to see the impact of different levels iron on mitigation of chlorosis in varagu, Paspalum scrobiculatum under calcareous soil and to investigate the influence of soil and foliar application of iron on growth, physiological and improvement of yield potential of varagu under calcareous soil condition. The varagu variety CO3 taken for this study The treatments comprised T1, NPK (44:22:0 kg ha-1) + 12.5 t  FYM/ha,T2, NPK (44:22:30 kg ha-1) +12.5 t FYM/ha, T3, T1 + Soil application of FeSO4   (25 kg ha-1), T4, T2 + Soil application of FeSO4 (25 kg ha-1), T5, T1 + Soil application of FeSO4 (50 kg ha-1), T6, T2 + Soil application of FeSO4 (50 kg ha-1), T7, T3 + Foliar spray of 0.5% FeSO4, T8, T4 + Foliar spray of 0.5% FeSO4, T9,T5 + Foliar spray of 0.5% FeSO4, T10,  T6 + Foliar spray of 0.5% FeSO4. During experimentation, morphological characteristics, growth attributes, physiological and biochemical components and biomass traits determined the mitigation of iron chlorosis. The iron deficiency in varagu was effectively controlled by T10, soil treatment 50 kg ha-1 FeSO4 and foliar spray of 0.5% FeSO4 applied on the 30th and 50th days after sowing through maintaining highest growth parameter values, maximum catalase and peroxidase activity and maintaining more chlorophyll content

Water Management in the Noyyal River Basin A Situation Analysis

The Noyyal sub-basin, which is 3510 sq km in area, is part of the Cauvery basin that lies in the state of Tamil Nadu. It is a rapidly urbanizing sub-basin that includes the Class I cities of Coimbatore and Tiruppur as well as 84 smaller urban settlements. Water issues in this basin have been the focus of much public debate and action over the last two decades. Most of the debate, triggered by farmer agitations and court cases, has focused on the question of water pollution; water scarcity and sustainability issues have received relatively little attention. Recent bans on industrial effluent discharge into the Noyyal, as well as changes in water supply infrastructure, watershed development activities, urban demand and agricultural water use have dramatically altered the future of the Noyyal River and merit follow-up studies. The purpose of this situation analysis is to summarize the current state of knowledge regarding water resources management in the Noyyal sub-basin and identify critical knowledge gaps to inform water-related research in the basin. It is hoped that such an analysis will help those studying or working on water issues in the Noyyal, and also provide useful insights for other urbanizing basins

Magnetic enhancement of Co0.2_{0.2}Zn0.8_{0.8}Fe2_2O4_4 spinel oxide by mechanical milling

We report the magnetic properties of mechanically milled Co$_{0.2}$Zn$_{0.8}$Fe$_2$O$_4$ spinel oxide. After 24 hours milling of the bulk sample, the XRD spectra show nanostructure with average particle size $\approx$ 20 nm. The as milled sample shows an enhancement in magnetization and ordering temperature compared to the bulk sample. If the as milled sample is annealed at different temperatures for the same duration, recrystallization process occurs and approaches to the bulk structure on increasing the annealing temperatures. The magnetization of the annealed samples first increases and then decreases. At higher annealing temperature ($\sim$ 1000$^{0}$C) the system shows two coexisting magnetic phases {\it i.e.}, spin glass state and ferrimagnetic state, similar to the as prepared bulk sample. The room temperature M\"{o}ssbauer spectra of the as milled sample, annealed at 300$^{0}$C for different durations (upto 575 hours), suggest that the observed change in magnetic behaviour is strongly related with cations redistribution between tetrahedral (A) and octahedral (O) sites in the spinel structure. Apart from the cation redistribution, we suggest that the enhancement of magnetization and ordering temperature is related with the reduction of B site spin canting and increase of strain induced anisotropic energy during mechanical milling.Comment: 14 pages LaTeX, 10 ps figure

Effect of operating conditions on yield and quality of biocrude during hydrothermal liquefaction of halophytic microalga Tetraselmis sp.

Abstract not availableB.E. Eboibi, D.M. Lewis, P.J. Ashman, S. Chinnasam

Crystallographically oriented magnetic ZnFe2O4 nanoparticles synthesized by Fe implantation into ZnO

In this paper, a correlation between structural and magnetic properties of Fe implanted ZnO is presented. High fluence Fe^+ implantation into ZnO leads to the formation of superparamagnetic alpha-Fe nanoparticles. High vacuum annealing at 823 K results in the growth of alpha-Fe particles, but the annealing at 1073 K oxidized the majority of the Fe nanoparticles. After a long term annealing at 1073 K, crystallographically oriented ZnFe2O4 nanoparticles were formed inside ZnO with the orientation relationship of ZnFe2O4(111)[110]//ZnO(0001)[1120]. These ZnFe2O4 nanoparticles show a hysteretic behavior upon magnetization reversal at 5 K.Comment: 21 pages, 7 figures, accepted by J. Phys. D: Appl. Phy

Mössbauer and magnetic study of Co x Fe3−x O4 nanoparticles

Magnetic nanoparticles of cobalt ferrites Co x Fe3−x O4 (x = 1 or 2) have been obtained either by mechanical milling or thermal treatment of pre-prepared layered double hydroxide carbonate x-LDH–CO3. Mechanical milling of the 1-LDH–CO3 leads to the large-scale preparation of nearly spherical nanoparticles of CoFe2O4, the size of which (5 to 20 nm) is controlled by the treatment time. Core-shell structure with surface spin-canting has been considered for the nanoparticles formed to explain the observed hysteresis loop shift (from ZFC–FC) in the magnetic properties. Annealing treatment of the 2-LDH–CO3 below 673 K results in the formation of nearly spherical pure Co2FeO4 nanoparticles. At 673 K and above, the LDH decomposition leads to the formation of a mixture of both spinels phases Co2FeO4 and CoFe2O4, the amount of the latter increases with annealing temperature. Unusually high magnetic hardness characterized by a 22 kOe coercive field at 1.8 K has been observed, which reflects the high intrinsic anisotropy for Co2FeO4

Assessment of rice (Co 51) seed ageing through volatile organic compound analysis using Headspace-Solid Phase Micro Extraction/ Gas Chromatography-Mass Spectrometry (HS-SPME/GCMS)

Seed ageing is an inevitable process that reduces seed quality during storage. When seeds deteriorate as a result of the lipid peroxidation process, it leads to produce toxic volatile organic compounds. These volatiles served as an indicator for the viability of stored seeds. With this background, the study was conducted to profile the volatile organic compounds emitted from rice seeds during storage. Volatile profiling of stored rice var. Co 51 seeds was done through Headspace-Solid phase microextraction/ Gas chromatography-mass spectrometry (HS-SPME/GCMS). The study clearly demonstrated that the significant decrease in physiological and biochemical quality attributes was noted due to an increase in the strength of volatiles released during ageing. When the release of total volatile strength reached more than 40%, a significant reduction in physiological attributes such as germination, root and shoot length, dry matter production and vigour index were observed. With respect to biochemical properties, a significant increase in electrical conductivity of seed leachate, lipid peroxidation and lipoxygenase activity, and decrease in dehydrogenase, catalase and peroxidase activities were observed. However, the highest reduction in all these properties were recorded when the total volatile strength reached to 54.90%. Finally, the study concluded that, among all the volatiles, 1-hexanol, 1-butanol, ethanol, hexanal, acetic acid, hexanoic acid and methyl ester were the most closely associated volatiles with seed deterioration. It indicates that these components could be considered the signature components for assessing the seed quality in rice during storage.

Size dependent magnetic properties and cation inversion in chemically synthesized MnFe2O4 nanoparticles

MnFe2O4nanoparticles with diameters ranging from about 4to50nm were synthesized using a modified coprecipitation method. X-ray diffractograms revealed a pure phase spinel ferrite structure for all samples. Transmission electron microscopy showed that the particles consist of a mixture of both spherical (smaller) and cubic (larger) particles dictated by the reaction kinetics. The Néel temperatures (TN) of MnFe2O4 for various particle sizes were determined by using high temperature magnetometry. The ∼4nm MnFe2O4 particles showed a TN of about 320°C whereas the ∼50nm particles had a TN of about 400°C. The high Néel temperature, compared with the bulk MnFe2O4 TN of 300°C, is due to a change in cation distribution between the tetrahedral and octahedral sites of the spinel lattice. Results of extended x-ray absorption fine structure measurements indicate a systematic change in the cation distribution dependent on processing conditions

Changes in daily mental health service use and mortality at the commencement and lifting of COVID-19 'lockdown' policy in 10 UK sites: a regression discontinuity in time design.

OBJECTIVES: To investigate changes in daily mental health (MH) service use and mortality in response to the introduction and the lifting of the COVID-19 'lockdown' policy in Spring 2020. DESIGN: A regression discontinuity in time (RDiT) analysis of daily service-level activity. SETTING AND PARTICIPANTS: Mental healthcare data were extracted from 10 UK providers. OUTCOME MEASURES: Daily (weekly for one site) deaths from all causes, referrals and discharges, inpatient care (admissions, discharges, caseloads) and community services (face-to-face (f2f)/non-f2f contacts, caseloads): Adult, older adult and child/adolescent mental health; early intervention in psychosis; home treatment teams and liaison/Accident and Emergency (A&E). Data were extracted from 1 Jan 2019 to 31 May 2020 for all sites, supplemented to 31 July 2020 for four sites. Changes around the commencement and lifting of COVID-19 'lockdown' policy (23 March and 10 May, respectively) were estimated using a RDiT design with a difference-in-difference approach generating incidence rate ratios (IRRs), meta-analysed across sites. RESULTS: Pooled estimates for the lockdown transition showed increased daily deaths (IRR 2.31, 95% CI 1.86 to 2.87), reduced referrals (IRR 0.62, 95% CI 0.55 to 0.70) and reduced inpatient admissions (IRR 0.75, 95% CI 0.67 to 0.83) and caseloads (IRR 0.85, 95% CI 0.79 to 0.91) compared with the pre lockdown period. All community services saw shifts from f2f to non-f2f contacts, but varied in caseload changes. Lift of lockdown was associated with reduced deaths (IRR 0.42, 95% CI 0.27 to 0.66), increased referrals (IRR 1.36, 95% CI 1.15 to 1.60) and increased inpatient admissions (IRR 1.21, 95% CI 1.04 to 1.42) and caseloads (IRR 1.06, 95% CI 1.00 to 1.12) compared with the lockdown period. Site-wide activity, inpatient care and community services did not return to pre lockdown levels after lift of lockdown, while number of deaths did. Between-site heterogeneity most often indicated variation in size rather than direction of effect. CONCLUSIONS: MH service delivery underwent sizeable changes during the first national lockdown, with as-yet unknown and unevaluated consequences