A practical guide to single-cell RNA-sequencing for biomedical research and clinical applications.

RNA sequencing (RNA-seq) is a genomic approach for the detection and quantitative analysis of messenger RNA molecules in a biological sample and is useful for studying cellular responses. RNA-seq has fueled much discovery and innovation in medicine over recent years. For practical reasons, the technique is usually conducted on samples comprising thousands to millions of cells. However, this has hindered direct assessment of the fundamental unit of biology-the cell. Since the first single-cell RNA-sequencing (scRNA-seq) study was published in 2009, many more have been conducted, mostly by specialist laboratories with unique skills in wet-lab single-cell genomics, bioinformatics, and computation. However, with the increasing commercial availability of scRNA-seq platforms, and the rapid ongoing maturation of bioinformatics approaches, a point has been reached where any biomedical researcher or clinician can use scRNA-seq to make exciting discoveries. In this review, we present a practical guide to help researchers design their first scRNA-seq studies, including introductory information on experimental hardware, protocol choice, quality control, data analysis and biological interpretation

Lessons from non-canonical splicing

Recent improvements in experimental and computational techniques that are used to study the transcriptome have enabled an unprecedented view of RNA processing, revealing many previously unknown non-canonical splicing events. This includes cryptic events located far from the currently annotated exons and unconventional splicing mechanisms that have important roles in regulating gene expression. These non-canonical splicing events are a major source of newly emerging transcripts during evolution, especially when they involve sequences derived from transposable elements. They are therefore under precise regulation and quality control, which minimizes their potential to disrupt gene expression. We explain how non-canonical splicing can lead to aberrant transcripts that cause many diseases, and also how it can be exploited for new therapeutic strategies

Printing in a Bind

Photographic representation of the art of printing a book in Frome Introduction: Camilla Brown Photographers: Charlotte Ball Michael Bradley [contributed eighteen photographs] Tereza Cervenova Johannah Churchill Rachel Gaon Oliver Garner Tricia Hague Patrick Kelly Becci Lee Erik Lovold Aleksandra Wojci

Effect of probiotic supplementation on immunoglobulins, isoagglutinins and antibody response in children of low socio-economic status

Background: Antigen exposure is one of the major exogenous factors modulating human immunocompetence acquisition. Decline in family size and improvements in public health and hygiene in developed countries, may deprive the immune system of appropriate antigen input by diminishing infectious stimuli. Probiotics are a large group of microorganisms defined by their beneficial effects on human health and with stimulating effects on different functions of the immune system. Aim of the study: We conducted a double-blind, placebo-controlled trial to determine if probiotics maintain their immune-stimulating effects in a population of 162 children with a high index of natural exposure to microorganisms. Children were to ingest for at least 4 months one of two products, low-fat milk fermented by Streptococcus thermophilus (control product) or low-fat milk fermented by S. thermophilus and Lactobacillus casei, with Lactobacillus acidophilus, oligofructose and inulin added after the fermentation process (test product). According to their age, children were vaccinated with DTP-Hib vaccine or a 23-valent anti-pneumococcal vaccine. Results: Final analysis of results was done in 70 children in each group, showing that the rate of immunoglobulin and isoagglutinin acquisition was similar in both groups. There was no difference between groups in antibody levels neither before nor after vaccination. Days of fever and number of episodes of infection were not statistically different in either group. Conclusions: Supplementation of standard fermented milk with additional probiotics was not of benefit. The high natural rate of early microbial exposure in infants and children from a population of low socio-economic status living in a "less hygienic environment" may account for the absence of an additional immune-stimulating effect by supplementary probiotics.Fil: Pérez, Néstor. Comisión de Investigaciones Científicas de la Provincia de Buenos Aires. Hospital de Niños de la Plata. Unidad de Inmunología; ArgentinaFil: Iannicelli, Juan C.. Comisión de Investigaciones Científicas de la Provincia de Buenos Aires. Hospital de Niños de la Plata. Unidad de Inmunología; ArgentinaFil: Girard-Bosch, Cecilia. Comisión de Investigaciones Científicas de la Provincia de Buenos Aires. Hospital de Niños de la Plata. Unidad de Inmunología; ArgentinaFil: Gonzalez, Silvia Nelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; ArgentinaFil: Varea, Ana. Comisión de Investigaciones Científicas de la Provincia de Buenos Aires. Hospital de Niños de la Plata. Unidad de Inmunología; ArgentinaFil: Disalvo, Liliana. Comisión de Investigaciones Científicas de la Provincia de Buenos Aires. Hospital de Niños de la Plata. Unidad de Inmunología; ArgentinaFil: Apezteguia, María. Comisión de Investigaciones Científicas de la Provincia de Buenos Aires. Hospital de Niños de la Plata. Unidad de Inmunología; ArgentinaFil: Pernas, Juan. Comisión de Investigaciones Científicas de la Provincia de Buenos Aires. Hospital de Niños de la Plata. Unidad de Inmunología; ArgentinaFil: Vicentin, Dimas. Sancor CUL. Departamento de Investigación, Innovación y Desarrollo; ArgentinaFil: Cravero, Ricardo. Sancor CUL. Departamento de Investigación, Innovación y Desarrollo; Argentin