Correlation of mRNA delivery timing and protein expression in lipid-based transfection

Non-viral gene delivery is constrained by the dwell time that most synthetic nucleic acid nanocarriers spend inside endosomal compartments. In order to overcome this endosomal-release bottleneck, methods are required that measure nanocarrier uptake kinetics and transfection efficiency simultaneously. Here, we employ live-cell imaging on single-cell arrays (LISCA) to study the delivery-time distribution of lipid-based mRNA complexes under varied serum conditions. By fitting a translation-maturation model to hundreds of individual eGFP reporter fluorescence time courses, the protein expression onset times and the expression rates after transfection are determined. Using this approach, we find that delivery timing and protein expression rates are not intrinsically correlated at the single-cell level, even though population-averaged values of both parameters conjointly change as a function of increasing external serum protein fraction. Lipofectamine-mediated delivery showed decreased transfection efficiency and longer delivery times with increasing serum protein concentration. This is in contrast to ionizable lipid nanoparticle (i-LNP)-mediated transfer, which showed increased efficiency and faster uptake in the presence of serum. In conclusion, the interdependences of single-cell expression rates and onset timing provide additional clues on uptake and release mechanisms, which are useful for improving nucleic acid delivery

A study of knowledge beliefs and attitudes regarding aids and human sexuality among medical college, engineering college and university Undergraduates of gorakhpur.

Research Problem: i) What is the level of knowledge and altitude of undergraduates about AIDS and human sexuality? ii) What arc the preferred modes of obtaining such knowledge?. Objectives: To assess the knowledge, beliefs and attitudes of undergraduate students regarding AIDS and human sexuality. Study Design: Self administered questionnaire. Setting and Participants: 1289 undergraduates from B.R.D. Medical College., M. M. M. Engineering College and University of Gorakhpur. Study Variables: Knowledge, beliefs and attitudes regarding AIDS and sexuality. Outcome Variables: Proportion of students having correct knowledge and positive attitudes. Statistical Analysis: By proportions. Result: l.evcl of knowledge about AIDS was generally high. Most of the students obtained knowledge about it through mass media. Few students had misconceptions about transmission of 1IIV infection. Knowledge about sex was obtained mainly from friends (36%) and books (31.31%). Most of the students preferred doctors (44.15%) and friends (43.66%) for asking something about sex. and not their parents (4.37%) or teachers (4.61%). 59.13% of boys and 34.49% of girls thought that students of their age had sex. Conclusion and Recommendations: The most peculiar fact in (his study is that students have no reliable means of obtaining correct information about subjects related to sex. Medical profession contributed very little in providing such knowledge. Most of them relied on their friends for such information. So. emphasis is to be given on recommending proper education material for the youth

The global impact of tobacco control policies on smokeless tobacco use: a systematic review

BACKGROUND: Smokeless tobacco, used by more than 300 million people globally, results in substantial morbidity and mortality. For smokeless tobacco control, many countries have adopted policies beyond the WHO Framework Convention on Tobacco Control, which has been instrumental in reducing smoking prevalence. The impact of these policies (within and outside the Framework Convention on Tobacco Control) on smokeless tobacco use remains unclear. We aimed to systematically review policies that are relevant to smokeless tobacco and its context and investigate their impact on smokeless tobacco use. METHODS: In this systematic review, we searched 11 electronic databases and grey literature between Jan 1, 2005, and Sept 20, 2021, in English and key south Asian languages, to summarise smokeless tobacco policies and their impact. Inclusion criteria were all types of studies on smokeless tobacco users that mentioned any smokeless tobacco relevant policies since 2005, except systematic reviews. Policies issued by organisations or private institutions were excluded as well as studies on e-cigarettes and Electronic Nicotine Delivery System except where harm reduction or switching were evaluated as a tobacco cessation strategy. Two reviewers independently screened articles, and data were extracted after standardisation. Quality of studies was appraised using the Effective Public Health Practice Project's Quality Assessment Tool. Outcomes for impact assessment included smokeless tobacco prevalence, uptake, cessation, and health effects. Due to substantial heterogeneity in the descriptions of policies and outcomes, data were descriptively and narratively synthesised. This systematic review was registered in PROSPERO (CRD42020191946). FINDINGS: 14 317 records were identified, of which 252 eligible studies were included as describing smokeless tobacco policies. 57 countries had policies targeting smokeless tobacco, of which 17 had policies outside the Framework Convention on Tobacco Control for smokeless tobacco (eg, spitting bans). 18 studies evaluated the impact, which were of variable quality (six strong, seven moderate, and five weak) and reported mainly on prevalence of smokeless tobacco use. The body of work evaluating policy initiatives based on the Framework Convention on Tobacco Control found that these initiatives were associated with reductions in smokeless tobacco prevalence of between 4·4% and 30·3% for taxation and 22·2% and 70·9% for multifaceted policies. Two studies evaluating the non-Framework policy of sales bans reported significant reductions in smokeless tobacco sale (6·4%) and use (combined sex 17·6%); one study, however, reported an increased trend in smokeless tobacco use in the youth after a total sales ban, likely due to cross-border smuggling. The one study reporting on cessation found a 13·3% increase in quit attempts in individuals exposed (47·5%) to Framework Convention on Tobacco Control policy: education, communication, training, and public awareness, compared with non-exposed (34·2%). INTERPRETATION: Many countries have implemented smokeless tobacco control policies, including those that extend beyond the Framework Convention on Tobacco Control. The available evidence suggests that taxation and multifaceted policy initiatives are associated with meaningful reductions in smokeless tobacco use. FUNDING: UK National Institute for Health Research

A Grassmann integral equation

The present study introduces and investigates a new type of equation which is called Grassmann integral equation in analogy to integral equations studied in real analysis. A Grassmann integral equation is an equation which involves Grassmann integrations and which is to be obeyed by an unknown function over a (finite-dimensional) Grassmann algebra G_m. A particular type of Grassmann integral equations is explicitly studied for certain low-dimensional Grassmann algebras. The choice of the equation under investigation is motivated by the effective action formalism of (lattice) quantum field theory. In a very general setting, for the Grassmann algebras G_2n, n = 2,3,4, the finite-dimensional analogues of the generating functionals of the Green functions are worked out explicitly by solving a coupled system of nonlinear matrix equations. Finally, by imposing the condition G[{\bar\Psi},{\Psi}] = G_0[{\lambda\bar\Psi}, {\lambda\Psi}] + const., 0<\lambda\in R (\bar\Psi_k, \Psi_k, k=1,...,n, are the generators of the Grassmann algebra G_2n), between the finite-dimensional analogues G_0 and G of the (``classical'') action and effective action functionals, respectively, a special Grassmann integral equation is being established and solved which also is equivalent to a coupled system of nonlinear matrix equations. If \lambda \not= 1, solutions to this Grassmann integral equation exist for n=2 (and consequently, also for any even value of n, specifically, for n=4) but not for n=3. If \lambda=1, the considered Grassmann integral equation has always a solution which corresponds to a Gaussian integral, but remarkably in the case n=4 a further solution is found which corresponds to a non-Gaussian integral. The investigation sheds light on the structures to be met for Grassmann algebras G_2n with arbitrarily chosen n.Comment: 58 pages LaTeX (v2: mainly, minor updates and corrections to the reference section; v3: references [4], [17]-[21], [39], [46], [49]-[54], [61], [64], [139] added

Population policies and education: exploring the contradictions of neo-liberal globalisation

The world is increasingly characterised by profound income, health and social inequalities (Appadurai, 2000). In recent decades development initiatives aimed at reducing these inequalities have been situated in a context of increasing globalisation with a dominant neo-liberal economic orthodoxy. This paper argues that neo-liberal globalisation contains inherent contradictions regarding choice and uniformity. This is illustrated in this paper through an exploration of the impact of neo-liberal globalisation on population policies and programmes. The dominant neo-liberal economic ideology that has influenced development over the last few decades has often led to alternative global visions being overlooked. Many current population and development debates are characterised by polarised arguments with strongly opposing aims and views. This raises the challenge of finding alternatives situated in more middle ground that both identify and promote the socially positive elements of neo-liberalism and state intervention, but also to limit their worst excesses within the population field and more broadly. This paper concludes with a discussion outling the positive nature of middle ground and other possible alternatives

Epidemiology of Knee and Hip Arthroplasty: A Systematic Review§

We present a systematic review of epidemiologic studies of Total Knee Arthroplasty (TKA) and Total Hip Arthroplasty (THA). The studies summarized in this systematic review provide us with estimates of arthroplasty utilization rates, underlying disease frequency and its trends and differences in utilization rates by age, gender and ethnicity among other factors. Among these, many studies are registry-based that assessed utilization rates using data from major orthopedic centers that may provide some understanding of underlying diagnosis and possibly time-trends. Several studies are population-based cross-sectional, which provide estimates of prevalence of TKA and THA. Population-based cohort studies included in this review provide the best estimates of incidence and utilization rates, time-trends and differences in these rates by important patient characteristics (age, gender, ethnicity and others). This article reviews the current published literature in the area and highlights the main findings

Evaluation of chemiluminescence, toluidine blue and histopathology for detection of high risk oral precancerous lesions: A cross-sectional study

<p>Abstract</p> <p>Background</p> <p>Early detection holds the key to an effective control of cancers in general and of oral cancers in particular. However, screening procedures for oral cancer are not straightforward due to procedural requirements as well as feasibility issues, especially in resource-limited countries.</p> <p>Methods</p> <p>We conducted a cross-sectional study to compare the performance of chemiluminescence, toluidine blue and histopathology for detection of high-risk precancerous oral lesions. We evaluated 99 lesions from 55 patients who underwent chemiluminescence and toluidine blue tests along with biopsy and histopathological examination. We studied inter-as well as intra-rater agreement in the histopathological evaluation and then using latent class modeling, we estimated the operating characteristics of these tests in the absence of a reference standard test.</p> <p>Results</p> <p>There was a weak inter-rater agreement (kappa < 0.15) as well as a weak intra-rater reproducibility (Pearson's r = 0.28, intra-class correlation rho = 0.03) in the histopathological evaluation of potentially high-risk precancerous lesions. When compared to histopathology, chemiluminescence and toluidine blue retention had a sensitivity of 1.00 and 0.59, respectively and a specificity of 0.01 and 0.79, respectively. However, latent class analysis indicated a low sensitivity (0.37) and high specificity (0.90) of histopathological evaluation. Toluidine blue had a near perfect high sensitivity and specificity for detection of high-risk lesions.</p> <p>Conclusion</p> <p>In our study, there was variability in the histopathological evaluation of oral precancerous lesions. Our results indicate that toluidine blue retention test may be better suited than chemiluminescence to detect high-risk oral precancerous lesions in a high-prevalence and low-resource setting like India.</p

Characterization of sequences in human TWIST required for nuclear localization

<p>Abstract</p> <p>Background</p> <p>Twist is a transcription factor that plays an important role in proliferation and tumorigenesis. Twist is a nuclear protein that regulates a variety of cellular functions controlled by protein-protein interactions and gene transcription events. The focus of this study was to characterize putative nuclear localization signals (NLSs) ³⁷RKRR⁴⁰and ⁷³KRGKK⁷⁷in the human TWIST (H-TWIST) protein.</p> <p>Results</p> <p>Using site-specific mutagenesis and immunofluorescences, we observed that altered TWIST^NLS1K38R, TWIST^NLS2K73R and K77R constructs inhibit nuclear accumulation of H-TWIST in mammalian cells, while TWIST^NLS2K76R expression was un-affected and retained to the nucleus. Subsequently, co-transfection of TWIST mutants K38R, K73R and K77R with E12 formed heterodimers and restored nuclear localization despite the NLSs mutations. Using a yeast-two-hybrid assay, we identified a novel TWIST-interacting candidate TCF-4, a basic helix-loop-helix transcription factor. The interaction of TWIST with TCF-4 confirmed using NLS rescue assays, where nuclear expression of mutant TWIST^NLS1with co-transfixed TCF-4 was observed. The interaction of TWIST with TCF-4 was also seen using standard immunoprecipitation assays.</p> <p>Conclusion</p> <p>Our study demonstrates the presence of two putative NLS motifs in H-TWIST and suggests that these NLS sequences are functional. Furthermore, we identified and confirmed the interaction of TWIST with a novel protein candidate TCF-4.</p