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Chromatin immunoprecipitation ChIP is the only molecular assay that directly determines, in a living cell, the binding association between a protein of interest and specific genomic loci. It is an indispensible tool in the biologist's toolbox, but the many limitations of this technique prevent broad adoption of ChIP in biological studies. The typical ChIP assay can take up to 1 wk to complete, and the process is technically tricky, yet tedious.

The ChIP assay yields are also low, thus requiring on the order of millions to billions of cells as starting material, which makes the assay unfeasible for studies using rare or precious samples. This protocol describes a microfluidics-based strategy for performing ChIP, which uses automation and scalability to reduce both total and hands-on assay time, and improve throughput. It allows whole fixed cells as input, and enables automated ChIP from as few as cells.

Antibody class switching is a feature of the adaptive immune system which enables diversification of the effector properties of antibodies. Even though class switching is essential for mounting a protective response to pathogens, the in vivo patterns and lineage characteristics of antibody class switching have remained uncharacterized in living humans. Here we comprehensively measured the landscape of antibody class switching in human adult twins using antibody repertoire sequencing.

The map identifies how antibodies of every class are created and delineates a two-tiered hierarchy of class switch pathways. Using somatic hypermutations as a molecular clock, we discovered that closely related B cells often switch to the same class, but lose coherence as somatic mutations accumulate. Such correlations between closely related cells exist when purified B cells class switch in vitro, suggesting that class switch recombination is directed toward specific isotypes by a cell-autonomous imprinted state.

The survival rate following lung transplantation is among the lowest of all solid-organ transplants, and current diagnostic tests often fail to distinguish between infection and rejection, the two primary posttransplant clinical complications. We describe a diagnostic assay that simultaneously monitors for rejection and infection in lung transplant recipients by sequencing of cell-free DNA cfDNA in plasma.

We determined that the levels of donor-derived cfDNA directly correlate with the results of invasive tests of rejection area under the curve 0. We also analyzed the nonhuman cfDNA as a hypothesis-free approach to test for infections. Cytomegalovirus is most frequently assayed clinically, and the levels of CMV-derived sequences in cfDNA are consistent with clinical results.

We furthermore show that hypothesis-free monitoring for pathogens using cfDNA reveals undiagnosed cases of infection, and that certain infectious pathogens such as human herpesvirus HHV 6, HHV-7, and adenovirus, which are not often tested clinically, occur with high frequency in this cohort.

Digital signaling enhances robustness of cellular decisions in noisy environments, but it is unclear how digital systems transmit temporal information about a stimulus. A sustained, weak stimulation lead to heterogeneous activation and delayed timing that is transmitted to gene expression.

In contrast, a transient, strong stimulus with the same area caused rapid and uniform dynamics. It remains difficult to predict and to measure the efficacy of pharmacological immunosuppression. We hypothesized that measuring the B-cell repertoire would enable assessment of the overall level of immunosuppression after heart transplantation. In this proof-of-concept study, we implemented a molecular-barcode-based immune repertoire sequencing assay that sensitively and accurately measures the isotype and clonal composition of the circulating B cell repertoire.

We used this assay to measure the temporal response of the B cell repertoire to immunosuppression after heart transplantation. We selected a subset of 12 participants from a larger prospective cohort study ClinicalTrials. This subset of 12 participants was selected to represent post-heart-transplant events, with and without acute rejection six participants with moderate-to-severe rejection and six without.

We analyzed samples from these patients, with an average follow-up period of 15 mo. To illustrate the potential of immune repertoire sequencing to monitor atypical post-transplant trajectories, we analyzed two more patients, one with chronic infections and one with amyloidosis.

A larger, prospective study will be needed to validate the power of immune repertoire sequencing to predict rejection events, as this proof-of-concept study is limited to a small number of patients who were selected based on several criteria including the availability of a large number of samples and the absence or presence of rejection events.

If confirmed in larger, prospective studies, the method described here has potential applications in the tailored management of post-transplant immunosuppression and, more broadly, as a method for assessing the overall activity of the immune system. Receptor tyrosine kinase RTK inhibitors have advanced colon cancer treatment. An array of patient-derived colon tumors and their associated xenografts were analyzed by immunohistochemistry to measure levels of KIT and its ligand KITLG.

Cells were analyzed by real-time quantitative reverse transcription polymerase chain reaction, single-cell gene expression analysis, flow cytometry, and immunohistochemical, immunoblot, and functional assays. Some mice were given the RTK inhibitor imatinib after injection of cancer cells; tumor growth was measured based on bioluminescence.

We assessed tumorigenicity using limiting dilution analysis. Knockdown of KIT decreased proliferation of colon cancer cell lines and growth of xenograft tumors in mice compared with control cells. KIT knockdown cells had increased expression of enterocyte markers, decreased expression of cycling genes, and, unexpectedly, increased expression of LGR5 associated genes.

Cancer cells with endogenous KIT expression were more tumorigenic in mice. KIT signaling promotes growth of colon cancer cells and organoids in culture and xenograft tumors in mice via its ligand, KITLG, in an autocrine or paracrine manner.

Like other structural variants, transposable element insertions can be highly polymorphic across individuals. Their functional impact, however, remains poorly understood. Current genome-wide approaches for genotyping insertion-site polymorphisms based on targeted or whole-genome sequencing remain very expensive and can lack accuracy, hence new large-scale genotyping methods are needed. We describe a high-throughput method for genotyping transposable element insertions and other types of structural variants that can be assayed by breakpoint PCR.

The method relies on next-generation sequencing of multiplex, site-specific PCR amplification products and read count-based genotype calls. We show that this method is flexible, efficient it does not require rounds of optimization , cost-effective and highly accurate. This method can benefit a wide range of applications from the routine genotyping of animal and plant populations to the functional study of structural variants in humans. The human brain is a tissue of vast complexity in terms of the cell types it comprises.

Conventional approaches to classifying cell types in the human brain at single cell resolution have been limited to exploring relatively few markers and therefore have provided a limited molecular characterization of any given cell type. We used single cell RNA sequencing on cells to capture the cellular complexity of the adult and fetal human brain at a whole transcriptome level.

Healthy adult temporal lobe tissue was obtained during surgical procedures where otherwise normal tissue was removed to gain access to deeper hippocampal pathology in patients with medical refractory seizures. We were able to classify individual cells into all of the major neuronal, glial, and vascular cell types in the brain. We were able to divide neurons into individual communities and show that these communities preserve the categorization of interneuron subtypes that is typically observed with the use of classic interneuron markers.

We then used single cell RNA sequencing on fetal human cortical neurons to identify genes that are differentially expressed between fetal and adult neurons and those genes that display an expression gradient that reflects the transition between replicating and quiescent fetal neuronal populations. Finally, we observed the expression of major histocompatibility complex type I genes in a subset of adult neurons, but not fetal neurons. The work presented here demonstrates the applicability of single cell RNA sequencing on the study of the adult human brain and constitutes a first step toward a comprehensive cellular atlas of the human brain.

It has long been thought that clonal deletion efficiently removes almost all self-specific T cells from the peripheral repertoire. For the Y chromosome-encoded SMCY antigen, self-specific T cells exhibited only a 3-fold lower average frequency in males versus females and were anergic with respect to peptide activation, although this inhibition could be overcome by a stronger stimulus.

We conclude that clonal deletion prunes but does not eliminate self-specific T cells and suggest that to do so would create holes in the repertoire that pathogens could readily exploit. In support of this hypothesis, we detected T cells specific for all 20 amino acid variants at the p5 position of a hepatitis C virus epitope in a random group of blood donors.

This protocol details a method for measuring the DNA methylation state of multiple target sites in single cells, otherwise known as single-cell restriction analysis of methylation SCRAM. The basic steps include isolating and lysing single cells, digesting genomic DNA with a methylation-sensitive restriction endonuclease MSRE and amplification of multiple targets by two rounds of PCR to determine the methylation status of target sites.

The method can reliably and accurately detect the methylation status of multiple target sites in each single cell, and it can be completed in a relatively short time View details for DOI Detecting tumor-derived cell-free DNA cfDNA in the blood of brain tumor patients is challenging, presumably owing to the blood-brain barrier. Cerebral spinal fluid CSF may serve as an alternative "liquid biopsy" of brain tumors by enabling measurement of circulating DNA within CSF to characterize tumor-specific mutations.

Many aspects about the characteristics and detectability of tumor mutations in CSF remain undetermined. Also, we applied cancer panel sequencing to globally characterize the somatic mutation profile from the CSF of 1 patient with suspected leptomeningeal disease. We detected tumor mutations in CSF samples from 6 of 7 patients with solid brain tumors.

The concentration of the tumor mutant alleles varied widely between patients, from View details for DOI Robotic and video game design challenges have been particularly effective in stimulating student interest, but equivalent experiences for the life sciences are not as developed.

Here we present the concept of a "biotic game design project" to motivate student learning at the interface of life sciences and device engineering as part of a cornerstone bioengineering devices course. We provide all course material and also present efforts in adapting the project's complexity to serve other time frames, age groups, learning focuses, and budgets. Students self-reported that they found the biotic game project fun and motivating, resulting in increased effort.

Hence this type of design project could generate excitement and educational impact similar to robotics and video games. Antibody heavy chains contain a variable and a constant region. The constant region of the antibody heavy chain is encoded by multiple groups of exons which define the isotype and therefore many functional characteristics of the antibody.

R1a-M is one of the most widely spread Y-chromosome haplogroups; however, its substructure within Europe and Asia has remained poorly characterized. The spatial frequency distributions of R1a sub-haplogroups conclusively indicate two major groups, one found primarily in Europe and the other confined to Central and South Asia.

Beyond the major European versus Asian dichotomy, we describe several younger sub-haplogroups. Based on spatial distributions and diversity patterns within the R1a-M clade, particularly rare basal branches detected primarily within Iran and eastern Turkey, we conclude that the initial episodes of haplogroup R1a diversification likely occurred in the vicinity of present-day Iran. RNA-protein interactions are central to biological regulation.

Cross-linking immunoprecipitation CLIP -seq is a powerful tool for genome-wide interrogation of RNA-protein interactomes, but current CLIP methods are limited by challenging biochemical steps and fail to detect many classes of noncoding and nonhuman RNAs. Here we present FAST-iCLIP, an integrated pipeline with improved CLIP biochemistry and an automated informatic pipeline for comprehensive analysis across protein coding, noncoding, repetitive, retroviral, and nonhuman transcriptomes.

Many cancers have substantial genomic heterogeneity within a given tumor, and to fully understand that diversity requires the ability to perform single cell analysis. We performed targeted sequencing of a panel of single nucleotide variants SNVs , deletions, and IgH sequences in 1, single tumor cells from six acute lymphoblastic leukemia ALL patients.

By accurately segregating groups of cooccurring mutations into distinct clonal populations, we identified codominant clones in the majority of patients. Evaluation of intraclonal mutation patterns identified clone-specific punctuated cytosine mutagenesis events, showed that most structural variants are acquired before SNVs, determined that KRAS mutations occur late in disease development but are not sufficient for clonal dominance, and identified clones within the same patient that are arrested at varied stages in B-cell development.

Taken together, these data order the sequence of genetic events that underlie childhood ALL and provide a framework for understanding the development of the disease at single-cell resolution. It has been postulated that there is a link between inflammation and cancer. Here we describe a role for cell-intrinsic toll-like receptor-2 TLR2; which is involved in inflammatory response signalling in normal intestinal and mammary epithelial cells and oncogenesis.

Limiting dilution transplantations of breast epithelial cells devoid of TLR2 or MYD88 revealed a significant decrease in mammary repopulating unit frequency compared with the control. These results suggest that inhibitors of the TLR2 pathway merit investigation as possible therapeutic and chemoprevention agents.

Homogeneous assay platforms for measuring protein-ligand interactions are highly valued due to their potential for high-throughput screening. However, the implementation of these multiplexed assays in conventional microplate formats is considerably expensive due to the large amounts of reagents required and the need for automation. We demonstrated the utility of this platform in determining the binding affinities between chromatin-regulatory proteins and different post-translationally modified histone peptides.

The microfluidic chip assay produces comparable results to conventional microtiter plate assays, yet requires 2 orders of magnitude less sample and an order of magnitude fewer pipetting steps. This approach enables one to use small samples for medium-scale screening and could ease the bottleneck of large-scale protein purification. Latent viral infection is a persistent cause of human disease. Although standard antiviral therapies can suppress active viral replication, no existing treatment can effectively eradicate latent infection and therefore a cure is lacking for many prevalent viral diseases.

Glaucoma is the second most common cause of blindness in the world. It is a multifactorial disease with several risk factors, of which intraocular pressure IOP is a primary contributing factor. IOP measurements are used for glaucoma diagnosis and patient monitoring. IOP has wide diurnal fluctuation and is dependent on body posture, so the occasional measurements done by the eye care expert in the clinic can be misleading. Here we show that microfluidic principles can be used to develop an implantable sensor that has a limit of detection of 1 mm Hg, high sensitivity and excellent reproducibility.

This device has a simple optical interface that enables IOP to be read with a smartphone camera. This sensor, with its ease of fabrication and simple design, as well as its allowance for IOP home monitoring, offers a promising approach for better care of patients with glaucoma. Monitoring allograft health is an important component of posttransplant therapy. Endomyocardial biopsy is the current gold standard for cardiac allograft monitoring but is an expensive and invasive procedure. Proof of principle of a universal, noninvasive diagnostic method based on high-throughput screening of circulating cell-free donor-derived DNA cfdDNA was recently demonstrated in a small retrospective cohort.

We present the results of a prospective cohort study 65 patients, samples that tested the utility of cfdDNA in measuring acute rejection after heart transplantation. Circulating cell-free DNA was purified from plasma and sequenced mean depth, 1. Through a comparison with endomyocardial biopsy results, we demonstrate that cfdDNA enables diagnosis of acute rejection after heart transplantation, with an area under the receiver operating characteristic curve of 0.

This noninvasive genome transplant dynamics approach is a powerful and informative method for routine monitoring of allograft health without incurring the risk, discomfort, and expense of an invasive biopsy. Insertions of the human-specific subfamily of LINE-1 L1 retrotransposon are highly polymorphic across individuals and can critically influence the human transcriptome. We hypothesized that L1 insertions could represent genetic variants determining important human phenotypic traits, and performed an integrated analysis of L1 elements and single nucleotide polymorphisms SNPs in several human populations.

We found that a large fraction of L1s were in high linkage disequilibrium with their surrounding genomic regions and that they were well tagged by SNPs. However, L1 variants were only partially captured by SNPs on standard SNP arrays, so that their potential phenotypic impact would be frequently missed by SNP array-based genome-wide association studies.

We next identified potential phenotypic effects of L1s by looking for signatures of natural selection linked to L1 insertions; significant extended haplotype homozygosity was detected around several L1 insertions. This finding suggests that some of these L1 insertions may have been the target of recent positive selection.

Circulating cell-free RNA in the blood provides a potential window into the health, phenotype, and developmental programs of a variety of human organs. We used high-throughput methods of RNA analysis such as microarrays and next-generation sequencing to characterize the global landscape circulating RNA in a cohort of human subjects.

By focusing on genes whose expression is highly specific to certain tissues, we were able to identify the relative contributions of these tissues to circulating RNA and to monitor changes in tissue development and health. As one application of this approach, we performed a longitudinal study on pregnant women and analyzed their combined cell-free RNA transcriptomes across all three trimesters of pregnancy and after delivery.

In addition to the analysis of mRNA, we observed and characterized noncoding species such as long noncoding RNA and circular RNA transcripts whose presence had not been previously observed in human plasma. We demonstrate that it is possible to track specific longitudinal phenotypic changes in both the mother and the fetus and that it is possible to directly measure transcripts from a variety of fetal tissues in the maternal blood sample.

We also studied the role of neuron-specific transcripts in the blood of healthy adults and those suffering from the neurodegenerative disorder Alzheimer's disease and showed that disease specific neural transcripts are present at increased levels in the blood of affected individuals. Characterization of the cell-free transcriptome in its entirety may thus provide broad insights into human health and development without the need for invasive tissue sampling.

The mammalian lung is a highly branched network in which the distal regions of the bronchial tree transform during development into a densely packed honeycomb of alveolar air sacs that mediate gas exchange. Although this transformation has been studied by marker expression analysis and fate-mapping, the mechanisms that control the progression of lung progenitors along distinct lineages into mature alveolar cell types are still incompletely known, in part because of the limited number of lineage markers and the effects of ensemble averaging in conventional transcriptome analysis experiments on cell populations.

Here we show that single-cell transcriptome analysis circumvents these problems and enables direct measurement of the various cell types and hierarchies in the developing lung. We used microfluidic single-cell RNA sequencing RNA-seq on individual cells at four different stages encompassing alveolar differentiation to measure the transcriptional states which define the developmental and cellular hierarchy of the distal mouse lung epithelium.

We empirically classified cells into distinct groups by using an unbiased genome-wide approach that did not require a priori knowledge of the underlying cell types or the previous purification of cell populations. The results confirmed the basic outlines of the classical model of epithelial cell-type diversity in the distal lung and led to the discovery of many previously unknown cell-type markers, including transcriptional regulators that discriminate between the different populations.

This single-cell genomics approach is applicable to any developing or mature tissue to robustly delineate molecularly distinct cell types, define progenitors and lineage hierarchies, and identify lineage-specific regulatory factors. Neurexins are evolutionarily conserved presynaptic cell-adhesion molecules that are essential for normal synapse formation and synaptic transmission.

Indirect evidence has indicated that extensive alternative splicing of neurexin mRNAs may produce hundreds if not thousands of neurexin isoforms, but no direct evidence for such diversity has been available. In addition, we observed several larger-scale events of alternative splicing that deleted multiple domains and were much less frequent than the canonical six sites of alternative splicing in neurexins. All of the six canonical events of alternative splicing appear to be independent of each other, suggesting that neurexins may exhibit an even larger isoform diversity than previously envisioned and comprise thousands of variants.

Efforts to determine the antibody repertoire encoded by B cells in the blood or lymphoid organs using high-throughput DNA sequencing technologies have been advancing at an extremely rapid pace and are transforming our understanding of humoral immune responses.

Information gained from high-throughput DNA sequencing of immunoglobulin genes Ig-seq can be applied to detect B-cell malignancies with high sensitivity, to discover antibodies specific for antigens of interest, to guide vaccine development and to understand autoimmunity. Rapid progress in the development of experimental protocols and informatics analysis tools is helping to reduce sequencing artifacts, to achieve more precise quantification of clonal diversity and to extract the most pertinent biological information.

That said, broader application of Ig-seq, especially in clinical settings, will require the development of a standardized experimental design framework that will enable the sharing and meta-analysis of sequencing data generated by different laboratories.

The biophysical details of how transcription factors and other proteins interact with RNA polymerase are of great interest as they represent the nexus of how structure and function interact to regulate gene expression in the cell. We used an in vitro microfluidic approach to map interactions between a set of ninety proteins, over a third of which are transcription factors, and each of the four subunits of E.

We detected interactions between RNA polymerase and transcription factors that earlier high-throughput screens missed; our results suggest that such interactions can occur without DNA mediation more commonly than previously appreciated. Single-cell sequencing is emerging as an important tool for studies of genomic heterogeneity. Whole genome amplification WGA is a key step in single-cell sequencing workflows and a multitude of methods have been introduced.

Here, we compare three state-of-the-art methods on both bulk and single-cell samples of E. We considered the effects of reaction gain on coverage uniformity, error rates and the level of background contamination. We compared the suitability of the different WGA methods for the detection of copy-number variations, for the detection of single-nucleotide polymorphisms and for de-novo genome assembly.

No single method performed best across all criteria and significant differences in characteristics were observed; the choice of which amplifier to use will depend strongly on the details of the type of question being asked in any given experiment. We present a method for obtaining long haplotypes, of over 3 kb in length, using a short-read sequencer, Barcode-directed Assembly for Extra-long Sequences BAsE-Seq. BAsE-Seq relies on transposing a template-specific barcode onto random segments of the template molecule and assembling the barcoded short reads into complete haplotypes.

We applied BAsE-Seq on mixed clones of hepatitis B virus and accurately identified haplotypes occurring at frequencies greater than or equal to 0. Applying BAsE-Seq to a clinical sample, we obtained over 9, viral haplotypes, which provided an unprecedented view of hepatitis B virus population structure during chronic infection.

BAsE-Seq is readily applicable for monitoring quasispecies evolution in viral diseases. There are few substantive methods to measure the health of the immune system, and the connection between immune strength and the viral component of the microbiome is poorly understood. Organ transplant recipients are treated with posttransplant therapies that combine immunosuppressive and antiviral drugs, offering a window into the effects of immune modulation on the virome.

We used sequencing of cell-free DNA in plasma to investigate drug-virome interactions in a cohort of organ transplant recipients samples, 96 patients and find that antivirals and immunosuppressants strongly affect the structure of the virome in plasma.

We observe marked virome compositional dynamics at the onset of the therapy and find that the total viral load increases with immunosuppression, whereas the bacterial component of the microbiome remains largely unaffected. The data provide insight into the relationship between the human virome, the state of the immune system, and the effects of pharmacological treatment and offer a potential application of the virome state to predict immunocompetence.

Epigenetic alterations are increasingly recognized as causes of human cancers and disease. These aberrations are likely to arise during genomic reprogramming in mammalian preimplantation embryos, when their epigenomes are most vulnerable. However, this process is only partially understood because of the experimental inaccessibility of early-stage embryos. Here, we introduce a methodologic advance, probing single cells for various DNA-methylation errors at multiple loci, to reveal failed maintenance of epigenetic mark results in chimeric mice, which display unpredictable phenotypes leading to developmental arrest.

Yet we show that mouse pronuclear transfer can be used to ameliorate such reprogramming defects. This study not only details the epigenetic reprogramming dynamics in early mammalian embryos but also suggests diagnostic and potential future therapeutic applications. Annual influenza vaccinations aim to protect against seasonal infections, and vaccine strain compositions are updated every year.

This protection is based on antibodies that are produced by either newly activated or memory B cells recalled from previous encounters with influenza vaccination or infection. The extent to which the B-cell repertoire responds to vaccination and recalls antibodies has so far not been analyzed at a genetic level-which is to say, at the level of antibody sequences. Here, we developed a consensus read sequencing approach that incorporates unique barcode labels on each starting RNA molecule.

These labels allow one to combine multiple sequencing reads covering the same RNA molecule to reduce the error rate to a desired level, and they also enable accurate quantification of RNA and isotype levels. We validated this approach and analyzed the differential response of the antibody repertoire to live-attenuated or trivalent-inactivated influenza vaccination. Additionally, we analyzed the antibody repertoire in response to repeated yearly vaccinations with trivalent-inactivated influenza vaccination.

We found antibody sequences that were present in both years, providing a direct genetic measurement of B-cell recall. Histocompatibility is the basis by which multicellular organisms of the same species distinguish self from nonself. Relatively little is known about the mechanisms underlying histocompatibility reactions in lower organisms. Botryllus schlosseri is a colonial urochordate, a sister group of vertebrates, that exhibits a genetically determined natural transplantation reaction, whereby self-recognition between colonies leads to formation of parabionts with a common vasculature, whereas rejection occurs between incompatible colonies.

Using genetically defined lines, whole-transcriptome sequencing, and genomics, we identified a single gene that encodes self-nonself and determines "graft" outcomes in this organism. These findings establish a platform for advancing the science of allorecognition. Library preparation for next-generation DNA sequencing NGS remains a key bottleneck in the sequencing process which can be relieved through improved automation and miniaturization. We describe a microfluidic device for automating laboratory protocols that require one or more column chromatography steps and demonstrate its utility for preparing Next Generation sequencing libraries for the Illumina and Ion Torrent platforms.

Sixteen different libraries can be generated simultaneously with significantly reduced reagent cost and hands-on time compared to manual library preparation. Using an appropriate column matrix and buffers, size selection can be performed on-chip following end-repair, dA tailing, and linker ligation, so that the libraries eluted from the chip are ready for sequencing.

The core architecture of the device ensures uniform, reproducible column packing without user supervision and accommodates multiple routine protocol steps in any sequence, such as reagent mixing and incubation; column packing, loading, washing, elution, and regeneration; capture of eluted material for use as a substrate in a later step of the protocol; and removal of one column matrix so that two or more column matrices with different functional properties can be used in the same protocol.

The microfluidic device is mounted on a plastic carrier so that reagents and products can be aliquoted and recovered using standard pipettors and liquid handling robots. The carrier-mounted device is operated using a benchtop controller that seals and operates the device with programmable temperature control, eliminating any requirement for the user to manually attach tubing or connectors.

In addition to NGS library preparation, the device and controller are suitable for automating other time-consuming and error-prone laboratory protocols requiring column chromatography steps, such as chromatin immunoprecipitation. High-throughput measurement of gene-expression and immune receptor repertoires have recently become powerful tools in the study of adaptive immune response.

However, despite their now-widespread use, both tend to discard cell identity by treating cell populations in bulk, and therefore lose the correlation between genetic variability and gene-expression at the single cell level. In order to recover this information, we developed a method to simultaneously measure gene expression profiles and genome mutations in single cells.

We applied this method by quantifying the relationships between gene expression and antibody mutation in ensembles of individual B-cells from immunized mice. The results reveal correlations reflecting the manner in which information propagates between a B-cell's antigen receptors, its gene expression, and its mutagenic machinery, and demonstrate the power of this approach to illuminate both heterogeneity and physiology in cell populations. OP9 is a yet-uncultivated bacterial lineage found in geothermal systems, petroleum reservoirs, anaerobic digesters and wastewater treatment facilities.

Here we use single-cell and metagenome sequencing to obtain two distinct, nearly complete OP9 genomes, one constructed from single cells sorted from hot spring sediments and the other derived from binned metagenomic contigs from an in situ-enriched cellulolytic, thermophilic community. Phylogenomic analyses support the designation of OP9 as a candidate phylum for which we propose the name 'Atribacteria'.

Although a plurality of predicted proteins is most similar to those from Firmicutes, the presence of key genes suggests a diderm cell envelope. Metabolic reconstruction from the core genome suggests an anaerobic lifestyle based on sugar fermentation by Embden-Meyerhof glycolysis with production of hydrogen, acetate and ethanol. Putative glycohydrolases and an endoglucanase may enable catabolism of hemi cellulose in thermal environments.

This study lays a foundation for understanding the physiology and ecological role of the 'Atribacteria'. The human antibody repertoire is one of the most important defenses against infectious disease, and the development of vaccines has enabled the conferral of targeted protection to specific pathogens. However, there are many challenges to measuring and analyzing the immunoglobulin sequence repertoire, including that each B cell's genome encodes a distinct antibody sequence, that the antibody repertoire changes over time, and the high similarity between antibody sequences.

We have addressed these challenges by using high-throughput long read sequencing to perform immunogenomic characterization of expressed human antibody repertoires in the context of influenza vaccination. Informatic analysis of 5 million antibody heavy chain sequences from healthy individuals allowed us to perform global characterizations of isotype distributions, determine the lineage structure of the repertoire, and measure age- and antigen-related mutational activity.

Our analysis of the clonal structure and mutational distribution of individuals' repertoires shows that elderly subjects have a decreased number of lineages but an increased prevaccination mutation load in their repertoire and that some of these subjects have an oligoclonal character to their repertoire in which the diversity of the lineages is greatly reduced relative to younger subjects. We have thus shown that global analysis of the immune system's clonal structure provides direct insight into the effects of vaccination and provides a detailed molecular portrait of age-related effects.

An apparatus that combines dynamic light scattering and Thioflavin T fluorescence detection is used to simultaneously probe fibril formation in polyglutamine peptides, the aggregating subunit associated with Huntington's disease, in vitro. Huntington's disease is a neurodegenerative disorder in a class of human pathologies that includes Alzheimer's and Parkinson's disease.

Despite the wide range of amino acid sequence in the aggregation prone polypeptides associated with these diseases, the resulting amyloids display strikingly similar physical structure, an observation which suggests a physical basis for amyloid fibril formation. The combined techniques allow elucidation of complex aggregation kinetics and are used to reveal multiple stages of amyloid fibril formation. Despite the enormous proliferation of bacterial genome data, surprisingly persistent collections of bacterial proteins have resisted functional annotation.

Many of these proteins are conserved across a large number of bacterial genomes. To assign a putative function to these conserved proteins of unknown function, we created a physical interaction map by measuring biophysical interaction of these proteins. Binary protein--protein interactions in the model organism Streptococcus pneumoniae TIGR4 are measured with a microfluidic high-throughput assay technology.

In some cases, informatic analysis was used to restrict the space of potential binding partners. In other cases, we performed in vitro proteome-wide interaction screens. We were able to assign putative functions to 50 conserved proteins of unknown function that we studied with this approach. In multicellular organisms and complex ecosystems, cells migrate in a social context. Whereas this is essential for the basic processes of life, the influence of neighboring cells on the individual remains poorly understood.

Previous work on isolated cells has observed a stereotypical migratory behavior characterized by short-time directional persistence with long-time random movement. We discovered a much richer dynamic in the social context, with significant variations in directionality, displacement, and speed, which are all modulated by local cell density.

We developed a mathematical model based on the experimentally identified "cellular traffic rules" and basic physics that revealed that these emergent behaviors are caused by the interplay of single-cell properties and intercellular interactions, the latter being dominated by a pseudopod formation bias mediated by secreted chemicals and pseudopod collapse following collisions. The model demonstrates how aspects of complex biology can be explained by simple rules of physics and constitutes a rapid test bed for future studies of collective migration of individual cells.

Multilayer microfluidics based on PDMS polydimethylsiloxane soft lithography have offered parallelism and integration for biological and chemical sciences, where reduction in reaction volume and consistency of controlled variables across experiments translate into reduced cost, increased quantity and quality of data. One issue with push up or push down microfluidic control concept is the inability to provide multiple control pressures without adding more complex and expensive external pressure controls.

We present here a microfluidic serial DAC Digital to Analog Converter that can be integrated with any PDMS device to expand the device's functionality by effectively adding an on-chip pressure regulator. The microfluidic serial DAC can be used with any incompressible fluids and operates in a similar fashion compared to an electronic serial DAC. It can be easily incorporated into any existing multilayer microfluidic devices, and the output pressure that the device generates could be held for extensive times.

We explore in this paper various factors that affect resolution, speed, and linearity of the DAC output. As an application, we demonstrate microfluidic DAC's ability for on-chip manipulation of flow resistance when integrated with a simple flow network. In addition, we illustrate an added advantage of using the microfluidic serial DAC in preventing back flow and possible contamination.

Genetic analysis of single cells is emerging as a powerful approach for studies of heterogeneous cell populations. Indeed, the notion of homogeneous cell populations is receding as approaches to resolve genetic and phenotypic variation between single cells are applied throughout the life sciences. A key step in single-cell genomic analysis today is the physical isolation of individual cells from heterogeneous populations, particularly microbial populations, which often exhibit high diversity.

Here, we detail the construction and use of instrumentation for optical trapping inside microfluidic devices to select individual cells for analysis by methods including nucleic acid sequencing. This approach has unique advantages for analyses of rare community members, cells with irregular morphologies, small quantity samples, and studies that employ advanced optical microscopy. Botryllus schlosseri is a colonial urochordate that follows the chordate plan of development following sexual reproduction, but invokes a stem cell-mediated budding program during subsequent rounds of asexual reproduction.

As urochordates are considered to be the closest living invertebrate relatives of vertebrates, they are ideal subjects for whole genome sequence analyses. Using a novel method for high-throughput sequencing of eukaryotic genomes, we sequenced and assembled Mbp of the B. A comparison of homologous genes between B. The B. The interaction of a comprehensive library of RNA mutants with stem-loop-binding protein precisely defined the RNA structural and sequence features that govern affinity.

The functional motif reconstructed in a single experiment on our platform uncovers new binding specificities and enriches interpretation of phylogenetic data. We determined a significant fraction of the genome sequence of a representative of Thiovulum, the uncultivated genus of colorless sulfur Epsilonproteobacteria, by analyzing the genome sequences of four individual cells collected from phototrophic mats from Elkhorn Slough, California.

These cells were isolated utilizing a microfluidic laser-tweezing system, and their genomes were amplified by multiple-displacement amplification prior to sequencing. Thiovulum is a gradient bacterium found at oxic-anoxic marine interfaces and noted for its distinctive morphology and rapid swimming motility. The genomic sequences of the four individual cells were assembled into a composite genome consisting of contigs covering 2.

This single-cell genome represents a genomic view of the physiological capabilities of isolated Thiovulum cells. This suggests that standard flagella are capable of propelling bacterial cells at speeds much faster than typically thought. Analysis of the genome suggests that naturally occurring Thiovulum populations are more diverse than previously recognized and that studies performed in the past probably address a wide range of unrecognized genotypic and phenotypic diversities of Thiovulum.

The genome presented in this article provides a basis for future isolation-independent studies of Thiovulum, where single-cell and metagenomic tools can be used to differentiate between different Thiovulum genotypes. The standard procedure to increase microfluidic chip performance is to grow the number of parallel test systems on the chip. This process is accompanied by miniaturizing biochemical workflows and micromechanical elements, which is often a major challenge for both engineering fields.

In this work, we show that it is possible to substantially increase the runtime performance of a microfluidic affinity assay for protein interactions by simultaneously engineering fluid logics and assay chemistry. For this, synergistic effects between the micro- and chemical architecture of the chip are exploited.

The presented strategy of reducing the runtime rather than size and volume of the mechanical elements and biological reagent compartments will, in general, be of importance for future analytical test systems on microfluidic chips to overcome performance barriers. Cystic fibrosis CF is an autosomal recessive disease caused by mutations in the gene encoding the CF transmembrane conductance regulator.

Disruption of electrolyte homeostasis at mucosal surfaces leads to severe lung, pancreatic, intestinal, hepatic, and reproductive abnormalities. Loss of lung function as a result of chronic lung disease is the primary cause of death from CF.

Using high-throughput sequencing to survey microbes in the sputum of 16 CF patients and 9 control individuals, we identified diverse microbial communities in the healthy samples, contravening conventional wisdom that healthy airways are not significantly colonized. Comparing these communities with those from the CF patients revealed significant differences in microbial ecology, including differential representation of uncultivated phylotypes.

Despite patient-specific differences, our analysis revealed a focal microbial profile characteristic of CF. The profile differentiated case and control groups even when classically recognized CF pathogens were excluded. As a control, lung explant tissues were also processed from a group of patients with pulmonary disease. The findings in lung tissue corroborated the presence of taxa identified in the sputum samples.

Comparing the sequencing results with clinical data indicated that diminished microbial diversity is associated with severity of pulmonary inflammation within our adult CF cohort. Given that most bone marrow cells are short-lived, the accumulation of multiple leukemogenic mutations in a single clonal lineage has been difficult to explain. We propose that serial acquisition of mutations occurs in self-renewing hematopoietic stem cells HSCs. We investigated this model through genomic analysis of HSCs from six patients with de novo acute myeloid leukemia AML.

Finally, through single-cell analysis, we determined that a clonal progression of multiple mutations occurred in the HSCs of some AML patients. These preleukemic HSCs suggest the clonal evolution of AML genomes from founder mutations, revealing a potential mechanism contributing to relapse. Such preleukemic HSCs may constitute a cellular reservoir that should be targeted therapeutically for more durable remissions.

Meiotic recombination and de novo mutation are the two main contributions toward gamete genome diversity, and many questions remain about how an individual human's genome is edited by these two processes. Here, we describe a high-throughput method for single-cell whole-genome analysis that was used to measure the genomic diversity in one individual's gamete genomes.

A microfluidic system was used for highly parallel sample processing and to minimize nonspecific amplification. High-density genotyping results from 91 single cells were used to create a personal recombination map, which was consistent with population-wide data at low resolution but revealed significant differences from pedigree data at higher resolution.

We used the data to test for meiotic drive and found evidence for gene conversion. High-throughput sequencing on 31 single cells was used to measure the frequency of large-scale genome instability, and deeper sequencing of eight single cells revealed de novo mutation rates with distinct characteristics. The vast majority of prenatal genetic testing requires invasive sampling.

However, this poses a risk to the fetus, so one must make a decision that weighs the desire for genetic information against the risk of an adverse outcome due to hazards of the testing process. These issues are not required to be coupled, and it would be desirable to discover genetic information about the fetus without incurring a health risk. Here we demonstrate that it is possible to non-invasively sequence the entire prenatal genome.

Our results show that molecular counting of parental haplotypes in maternal plasma by shotgun sequencing of maternal plasma DNA allows the inherited fetal genome to be deciphered non-invasively. We also applied the counting principle directly to each allele in the fetal exome by performing exome capture on maternal plasma DNA before shotgun sequencing. This approach enables non-invasive exome screening of clinically relevant and deleterious alleles that were paternally inherited or had arisen as de novo germline mutations, and complements the haplotype counting approach to provide a comprehensive view of the fetal genome.

Non-invasive determination of the fetal genome may ultimately facilitate the diagnosis of all inherited and de novo genetic disease. The root functions as the physical anchor of the plant and is the organ responsible for uptake of water and mineral nutrients such as nitrogen, phosphorus, sulfate and trace elements that plants acquire from the soil.

If we want to develop sustainable approaches to producing high crop yield, we need to better understand how the root develops, takes up a wide spectrum of nutrients, and interacts with symbiotic and pathogenic organisms. To accomplish these goals, we need to be able to explore roots in microscopic detail over time periods ranging from minutes to days. We developed the RootChip, a polydimethylsiloxane PDMS - based microfluidic device, which allows us to grow and image roots from Arabidopsis seedlings while avoiding any physical stress to roots during preparation for imaging 1 Figure 1.

The device contains a bifurcated channel structure featuring micromechanical valves to guide the fluid flow from solution inlets to each of the eight observation chambers 2. This perfusion system allows the root microenvironment to be controlled and modified with precision and speed. The volume of the chambers is approximately nl, thus requiring only minimal amounts of test solution. Here we provide a detailed protocol for studying root biology on the RootChip using imaging-based approaches with real time resolution.

Roots can be analyzed over several days using time lapse microscopy. Roots can be perfused with nutrient solutions or inhibitors, and up to eight seedlings can be analyzed in parallel. This system has the potential for a wide range of applications, including analysis of root growth in the presence or absence of chemicals, fluorescence-based analysis of gene expression, and the analysis of biosensors, e. FRET nanosensors 3. Segmented filamentous bacteria SFB are host-specific intestinal symbionts that comprise a distinct clade within the Clostridiaceae, designated Candidatus Arthromitus.

We are opening our church to serve tea and cakes to the wider community as well as members of our own congregation. We will have seating inside and outside the church and be decorating with bunting, balloons etc. On Saturday 4th June from 2 to 4 pm. We hope to raise funds by donation for […]. We are very grateful to Thames Water for the opportunity and hope that supporters of the Club will […]. Closing the High Street we will offer fun fair rides, street games, face painting, entertainment, food and […].

We are a local museum, and will be displaying artefacts from our collection of Royal memorabilia covering the last years. On Thursday 2nd June at 9. During the afternoon there will be games on the common including the pillow fight championships, egg throwing, welly wanging and various races and at 7. Our organisation is a local community based social club. There will be a Bouncy Castle and Face Painting for children. Non members are welcome to attend. Advance booking for the BBQ is recommended but […].

Free childrens entertainment, circus acts and workshop, fun fair, face […]. We are part of the national beacon lightings. This event is planned to be the biggest open-air event in Purfleet-on-Thames for over years, in fact since when Winston Churchill banned the Purfleet Fair by an Act […]. Join us for a lovely community picnic in the park. We will be encouraging families to get together over the half term, bank holiday and jubilee on Friday 3rd June from 10am. Our project enables the community to come together and connect socially and try new opportunities you may not have tried before.

Join us […]. Buy a ticket for a Hog Roast or bring your own picnic. Activities for the kids young and old refreshments and a whole load of community spirit. The Glebe Woldingham is a recreation space in the heart of Woldingham village on the Northdown in Surrey. No need to book, just drip in. Children under 8 must be accompanied by an adult. Call us on 41 31 31 or email templehilllibrary kent.

On Friday 3rd June we will have general open ringing from Free drop in Jubilee craft event throughout the day. Cake and tea offered throughout the day. Bring your own picnic for Open Mike sessions 12pm to 3pm with live music. Plan to have kids activities like a bouncy castle and games, we will all as a community contribute towards food. Music will be supplied. Thurrock libraries provides an exciting modern range of services including free internet access, free wi-fi, books, e-books, e-magazines and events for all ages.

Enjoy a relaxing evening pom pomming, and take a home a crown, party ready! All materials […]. Come and enjoy a glass of Pimms and a few nibbles — perhaps get your supper from The Frying Squad and eat it in the Village Hall or, if a nice evening, out on the green. Entertainment from the village choir, buglers and […]. On Thursday 2nd June, people from across the region are invited to join a Jubilee celebration like […]. Our minute concert is free to access, with a […].

Visitors on Sunday June 5 will be able to enjoy their own hampers on the famous parterre, in the Japanese Garden or by the lakeside, or purchase cake, refreshments or ice-creams […]. Come along and join the Jubilee picnic. There will also be craft activities and treasure hunt in library 31 May to 4 June. On Thursday June 2nd at 9. We will be hosting a Jubilee storyhunt with professional storyteller John KIrk.

We are a Parish Council and will be hosting a fete, picnic in the park, live music in the evening leading up to the lighting of the beacon at 9. There will be a baking competition, inflatables, races and lots of other stalls. Visitors can see an array of memorabilia on display and find out more about how locals celebrated royal events in the past.

Using textiles and film, the Seventy Sewn Stories exhibition will celebrate and present the lives and changes lived and seen by local people in Hoddesdon and the surrounding areas, from the last 70 years. Young people aged between 11 and 16 years and people of all ages, have worked collaboratively with artists and the Lowewood […]. The earliest photograph being of celebrations […]. Throughout half-term, there will be a Jubilee Trail to visit in the churchyard, fun for all ages!

Throughout half-term there will be a Jubilee Trail to visit in the churchyard, fun for all ages! Brooklands Museum is the home of British motorsport and aviation, with a rich history covering over years of British innovation. To celebrate the platinum jubilee we are hosting a royal trail for children to spot around the museum on their visit and earn a prize.

We will also have royal themed crafts and activities […]. Also archery, croquet and Union Jack face-painting for children. Addlestone has the fastest […]. Oxted Library Teddy Bears picnic free event — bring your own picnic lunch, teddy and enjoy our stories and songs. Ages 6 and under. Everyone can also play in a first timers free-to-enter bowls tournament or just come and spectate and munch cake.

Free drop-in craft event throughout the day, hosted by Home Instead. Cake and tea for sale throughout the day provided by Friends of Chertsey Library. Bring your own picnic for Open Mike sessions 12pm until 3pm with live music. Our office will be transformed into a festive party for the day to mark this once-in-a-lifetime moment, hosting a royal tea with recipes Her Majesty favours for her daily afternoon tea. As Royal tradition says our Afternoon Tea […]. On School Mead, Abbots Langley, from 12 noon to pm.

Free teas and squash. Bring your own picnic. Bring Cake to Share. Music from the past seven decades by British […]. No age restrictions apply. Sunday 5th […]. Birchwood Methodist Church is a church in Hatfield which welcomes all. It holds community events alongside regular Sunday morning worship at We shall be celebrating with a tea party on the lawn, games for adults and activities for children.

Friday 3rd June — 11am to 4pm — in the church grounds. More details to follow, […]. Serving the communities of Egham, Egham Hythe, Englefield Green, Thorpe and Virginia Water, we explore the history of the local area from its earliest […]. The parish council will be supplying tables and chairs, all are very welcome to bring a picnic and join in the celebrations with your Brasted neighbours.

We are expecting the Teashop opposite the Green will be open […]. Join us for a chat, a cup of tea and a slice of cake on Wednesday 1 June, 3pm to 4 pm. All are welcome to join us today and celebrate the Jubilee. Join us at Otford Library for a special Platinum Jubilee craft and storytime for pre-schoolers! No need to book, just drop in from 2 to 3pm. Call us on 41 31 31 or email otfordlibrary kent. A Turkish restaurant based in Brentwood, Essex. There will be live music from a local choir and a piper performing, as well as a speech from the newly sworn in Mayor of the borough.

Attendees are encouraged to arrive from 9pm, […]. The Otford Society represents all residents of this rural village. Supported by the Parish Council we are holding a big barbecue on the village green on Thursday 2nd June, starting at 12PM. There will be a hog-roast, hamburgers, sausage baps and vegan food. The Rising Sun pub will run the beer tent.

Ice cream, […]. The whole afternoon is free and accessible to all. The exhibition will open on Saturday, 28 May and run through to Friday, 24 June. We are holding a fun day staring at 1pm until 10pm, our aim is to come together as a community and celebrate this wonderful occasion.

We are ensuring there are is free entertainment, refreshments and fun along with some that you can buy. Large outdoor movie screens showing films, singer, community tea and cake, wacky […]. Our aim is to welcome everybody from the local community, to have a fun day.

Come along and enjoy an exciting afternoon of live entertainment, a delicious barbecue, drinks and cocktails in our lovely garden and communal lounge on the 2nd June. Everyone is welcome — friends, family and children.

Our friendly team will be […]. Call us on 41 31 31 or email longfieldlibrary kent. Join us from 7. Enjoy a relaxing morning pom pomming, and take a home a crown, party ready! Join us for a evening with the very talented Ed, join us for a meal and then dance the night away at our right royal knees up bash.

The programme includes music from the original coronation service alongside rousing and patriotic favourites from Hubert Parry, William Walton, Georg Frederic Handel and Henry Purcell, together with contemporary music by Will Todd and a brand […]. Make a Royal Crown Tuesday 31 May Slough Libraries will be hosting a range of events for all ages to mark this historic occasion.

Activities include story times and crafts, music performances and workshops with Royal Collection Trust. Supported by Arts Council England. Refreshments, shopping, activities, games and fun for all the family. Thursday evening 2nd June — Lighting of the Beacon as part of the national chain. Acrosphere display Friday 3rd June — Cricket in the Recreation Ground, followed by an open-air film show. BBQ and beer. Beacon lighting, live music and disco, evening picnic on the green. Food and refreshments will also be available to purchase or bring your own picnic.

We have hosted several ceramics workshops for adults and children in partnership with a local ceramics , Manic Ceramics, making and decorating ceramic crowns, corgis and bunting to hang on a tree sculpture and installed in our gardens […]. With live music on the historic terrace from 1pm-3pm, the south lawn is the perfect place to sit with friends and family with a picnic and enjoy the stunning views across the […].

Kent Wildlife Trust manage over 65 nature-rich reserves around the county. Our mission is to help organisations, people and communities connect to nature and care for our natural places. A large piece of this puzzle is encouraging future generations to enjoy and appreciate the wild spaces around them. To this end we are hosting a […].

After two years of Covid we are bringing the community together to celebrate the Queens Platinum Jubilee with a picnic on the Glebe Field and the lighting […]. We encourage anyone interested in amateur radio […]. Designed by the British couturier Sir Norman Hartnell, the dress was created in the finest white duchesse satin, richly embroidered in a lattice-work effect with an iconographic scheme of national […].

The Celebration will see more than horses and performers create a minute piece of spellbinding arena theatre that will include actors and artists, […]. Wateringbury Scouts are holding a family camp at Thristwood Scout campsite over the 4 day weekend. We will run games and activities to help celebrate this monumental milestone. We will […]. THIS […]. We will be hosting the event at the Theatre Royal in Windsor, Berkshire. The event is open for anyone to join and it is free to attend. The Pyrford and Wisley Village Show is celebrating its 75th year in In addition to the arena events and dog show there will Live music as well as all the usual attractions on the show ground, including a selection of stalls, […].

We are a Parish Council who will be supporting the community to hold a fun event at Salfords Social Club between midday and 4pm. We hope to bring the community together with a picnic, music and fun and games for all the family. Pop along to Seal Library on Saturday 4 June between 11am and pm and make a crown fit for a Queen! Suitable for all ages, children to be accompanied by a parent or carer. No need to book, just drop in from 11am.

Email seallibrary kent. Seal Know Your Neighbours exists to help members of our community facilitate community projects and events, such as lighting the village at Christmas, hosting community parties and celebrations and enhancing facilities for community organisations. Our Jubilee events include an exhibition of Seal over the past 70 years. East Horsley Parish Council acts as the official voice of the village, dealing with bodies such as Guildford Borough Council, Surrey County Council, Police, medical services, Surrey Highways, Network Rail, etc, to try and ensure an effective delivery of their services across the village.

East Horsley along with West Horsley will celebrate the 70th Platinum […]. The Big Community Sing is a fantastic event giving local singers the chance to perform before a packed-out theatre while raising money for local mental health support groups. There will be a further celebration on the 4th June pm on the ingrave playing fields with a marquee, live entertainment, kids activities, food and drinks will also be available.

Please speak to a member of staff for more details or visit www. Make a Royal Crown Monday 30 May The position of the beacon will be facing the Thames and in the location where the Tudor block house once stood. This 2-day traditional agricultural show is a large public event that celebrates life in the countryside every year, but specially for the Show will include a Jubilee Marquee filled with music and dancing from the s.

Hertfordshire Hearing Advisory Service supports people who are Deaf or affected by hearing loss. I made the crochet installation for the Platinum Jubilee of the queen. It is on dispaly on top of the post box in Park Road, Woking.

Photo attached. I have more pictures if you need them. Hope you like it. Best wishes, Carla. Thanksgiving service for the Queen 7. Entertainment throughout the day on Rainbow Stage with musicians and theatre performances. Activity corner with colouring in, face painting, balloon modelling and bubbles. Walkabout characters 9 hole Crazy Golf Giant games corner There will be deck chairs and additional tables and chairs throughout the town centre and some great Jubilee picnic lunch offers from various […].

As the sun sets on 2 June, join us in Jubilee Square in Woking for a unique opportunity to celebrate Her Majesty the Queen and her achievements. Members of frontline and emergency services are particularly invited to this event, to enable the community to publicly thank them for their dedication and hard work during the […].

Residents are invited to bring a picnic to The Green. No need to book, just drop in from 11am to 1pm. Call us on 41 31 31 or email edenbridgelibrary kent. Hertfordshire Libraries provides the local community with access to information in many formats, most notably books and computers.

Therefore we hold a large collection of stock relating to Queen Elizabeth and the Royal Family which will be on display during the Platinum Jubilee. The day will culminate in the lighting of the New village Jubilee Beacon and the singing […]. There will be stalls, a tea tent, BBQ, live music and Jubilee celebration activities. We are thrilled to announce that the event, based on Harpenden Common, will include live music and entertainment, food and drink stalls, outdoor tables and community seating, games, […].

Hosted by the Mayor of Harpenden, this will be a memorable and […]. The Dandelion Cafe is a monthly workshop for Older Adults in the Edenbridge area, offering creative activities and entertainment in an inclusive, supportive environment. We run on the first Wednesday of the month, pmpm.

Please join […]. We invite local residents to come along with their picnics and come along to Horley Recreation Ground for an afternoon of fun, entertainment and celebrations. Picnics will also be available […]. The event will be hosted by actor and comedian Andre Vincent. During the day there will be performances from Gobstoppers and Joanna […]. Stangrove Park will be alive with music from 7pm, and the […]. The event will celebrate the Platinum Jubilee with street and stage entertainment, stalls, a River of Hope procession, a Platinum Pudding Competition, food stalls, bar and bbq.

We will be hosting an exhibition and talk about the Queens connection to India and the Commonwealth. The event will begin at […]. Visit Laindon Community Centre […]. Cream Teas will be available in the Churchyard throughout the day! Make a Royal Crown Wednesday 1 June 2. Welcom forum are volunteers interested in making life better in the local community and liaise with many groups. They also try where possible to organise a local fun day and have initiated and look after some publically accessible AEDs in the area.

We engage with council and developers alike with our […]. Research and document local history, have visiting speakers on historic subjects, publish books on local history. A night of music reminiscent of the s at Sawbridgeworth Memorial Hall on Friday 3rd June — doors open 7pm.

Licensed bar. Take home a souvenir. Proceeds on the night […]. Sawbridgeworth Local History Society in conjunction with Sawbridgeworth Memorial Hall CM21 9AX will be holding an event on Friday 3rd June with a night of music reminiscent of the s with the trio The Decibelles and there will be a souvenir to take home. Doors open 7pm. Proceeds on the night donated to […].

After your walk, join us in our beautiful Hospice gardens where […]. The Steeple View Community Association is a local charity based in Laindon that organises lots of free community events to connect the whole community. On June 3rd at Steeple View Memorial Hall we are holding an afternoon tea party for families in the local community.

Food and entertainment is included. There will be lots of […]. Here is an opportunity to enjoy an evening of prom-style celebration at the beginning of the festival too! We are two local artists. To celebrate the Jubilee we are holding an exhibition and sale of our art, ceramics and crafts, many of which have a royal theme. There will be tea, cakes and other refreshments and a section selling saplings and seedlings as part of the Queens Green Canopy initiative.

We have pipers playing at 9. Visit Sun Corner on Thursday 2 June between pm to pm to see one […]. Visit The Paddocks on Thursday 2 June between pm to 10pm to see one […]. Visit The Paddocks on […]. We are bringing the village together to enjoy a lunch, raffle, competitions, fish and chips and each household will be gifted a tree to plant in commemoration of the day. The Oasis Partnership provides support, activities and opportunities to help individuals to improve their health and wellbeing.

The Oasis Art Box is our community arts project. The event will include a swing band, ice cream and cream teas and giant garden games. Working with community groups we will also be hosting a football competition and inflatable fun area. Over the weekend, there […].

Our plan is to hold an open air lunch in the Church grounds and invite our local residents to it. This will be an excellent opportunity for everyone to get to know their […]. Holyport Community Trust HCT is a charity that supports the community of Holyport via the use of a village hall that is hired out and used by HCT and hirers for many events of different types year round.

Visit Gloucester Park on Thursday 2 June between pm to 10pm to see one […]. Visit Gloucester Park on […]. Join your fellow wine lovers in our vineyard marquee to raise a glass to Queen Elizabeth II becoming the first British Monarch to celebrate a Platinum Jubilee, marking 70 years of service to the people of the United Kingdom and the Commonwealth.

Thanks to the help of one of our Kenelm Club members, Digby is […]. As loyal citizens of Britain, the Ahmadiyya Muslim Noor Mosque will be celebrating the Jubilee by holding a lunch, which will give the opportunity for friends, neighbours and members of the local community to take a tour around the Mosque. Lovely Jub i Lee: Come Together is a royally uplifting experience, promoting social cohesion through dancing, live music, feasting and making.

Each event will […]. Pirbright Parish Council is a local authority. PPC will host a beacon lighting event with live entertainment. PPC will host a Big Picnic on the village green, with entertainment and food stalls. PPC will be […]. We are celebrating the Queens Jubilee on Sunday 5th June from 1. BYO food and drinks and perhaps something for the sharing table. A fun community event for local residents. Come and join Guildford Library for Storytime and a picnic.

People can arrive from 2pm for the picnic lunch with the actual Storytime starting at 2. The fete is free on the gate and open to all Crawley residents, with access to the field via […]. Vitality Church is a church for the local community in south-east Bracknell. Guildford Museum celebrates the Platinum Jubilee with an exhibition of objects and images remembering how Guildford has celebrated royal occasions Historic photographs show how the town decorated itself for past jubilees and coronations.

Her Majesty Queen Elizabeth […]. As the sun sets on 2 June, join us at Guildford Cathedral for a unique opportunity to celebrate Her Majesty the Queen and her achievements. Spectators are invited to gather outside the front of the cathedral from 9. The evening will close with the lighting […].

A Jubilee Raffle will be held at 1. Weather permitting there will be a barbecue; otherwise food will be prepared in the […]. Commencing at 2. In the evening local musician John Knowles will provide entertainment. Food and refreshments will be available. Weather permitting there will be a barbecue; otherwise food will be prepared in the kitchen. Visit Chalvedon Community Centre […]. We have a beacon to be lit along with a piper playing during this event at the front […].

BBQ, sweet and savoury foods, range of music throughout afternoon and into evening, sporting activities, Bouncy Castle, stalls […]. There will be more than 50 street parties across the borough where you can get together with your neighbours, raise a glass and enjoy a great British tradition of marking special royal occasions.

At historic Tonbridge Castle […]. This has been organised to raise money […]. Our party will be an all-day event. Refreshments of wine and nibbles. Raffle tickets will be on sale. Booked tickets only. Please […]. The Marlow Bellringers ring the bells at All Saints, Marlow every Sunday morning to call worshippers to church and have done so since the 18th century when the current bells were installed. On occasions such as Coronations, Jubilees and Royal Weddings, the 8 bells are rung for a period of 3 hours in what is […].

We are thrilled to be involved in creating a community event in Sussex in celebration of the Queens jubilee. Singing mamas are a national movement for maternal wellbeing. We build communities or women and mothers who sing -to improve wellbeing in families. Our singing groups are for women and mothers to meet weekly, learning and […]. We are a family-run model railway shop based in Essex.

On Saturday, we will be holding a small open day in our shop with a special sale across our British items, as well as offering typical British refreshments to everyone. We will have an N Gauge layout set up, so the children can have a […]. We will be joining in the rest of the UK […]. Step back in time as we honour both our sovereign and arguably our greatest author at a weekend of Dickensian discovery with a Jubilee twist.

Immerse yourself in a cultural celebration of literature and entertainment and take in character parades from bygone years, eclectic live music performances, prose and poetry readings and comedy skits as […]. We track issues affecting our village and work closely with other organisations to influence local authorities and others to make our voice heard for the benefir of residents, businesses and visitors Village Fete is a family fun day which will showcase the fabulous talents of our voluntary organisations, schools and churches.

Live music plus streaming […]. This England is a Jubilee Celebration taking place at 7. Performed by Serendipity It will combine the best English prose, verse and song. We are holding a celebration BBQ on the evening of Friday 3rd June , this will also have live music from Miss Noma, a local band playing a great selection of covers. Thursday 2 June in The Square at Visit The Nevendon Centre […].

We will be running different Big Lunch sessions there are three time slot options to choose from 11am until We have arranged a village picnic with free food and refreshments for residents. All free tickets have now been allocated but anyone is welcome to join on the […]. The Platinum Jubilee Beacons is the first national event of the celebration weekend.

It will see thousands of communities, charities and different groups throughout the regions of the UK, Channel Islands, Isle of Man and UK Overseas Territories come together to light beacons at exactly the same time. A long-standing tradition at royal jubilees, weddings […].

ENTER: the pudding competition! Friends of Wandon Park Luton, are fighting to save our community park from being made into a housing development. We hold various community events throughout the year, and it is a much loved and well used park, a valued green space in the heart of our community. We will be beginning our celebrations by lighting our Beacon, which has been made by our Landlord. Throughout the afternoon, a procession of historic, commercial and leisure sea vessels will sail up-river to Sun Pier, Chatham, to join a synchronised spectacle of sights and sounds from ship to shore.

Throughout the afternoon, a procession of historic, commercial and leisure sea vessels will sail up-river. We will be lighting the fires and serving a delicious Hog Roast on our wonderful waterfront terrace. Our now Famous Tapas menu will be available and […].

This event is open to all and is FREE to attend. We have FREE face painting. Food and drink vendors, Cream Teas, Display of historic photographs, […]. We are having a village event. Live band and DJ, beer tent and bouncy castle. All race participants will receive a special commemorative medal. The race starts at 9am on the village green […]. Many of our Zoobilee activities are free with your entry ticket or membership card.

Under 3s go free. We will be serving refreshments, tea, coffee, cakes, and burgers at our coffee shop. Enjoy […]. There will be crown making, a Jubilee treasure hunt, and a Platinum Jubilee quiz. Head to kent. Live music, entertainment, rides, market stalls, refreshments stalls, licensed bar.

Local Shotgate Jubilee pudding competition. Advertising poster competition for local schools. Avoid single use plastic if […]. The Net Community Hub, in partnership with its befriending and volunteering group, Walderslade Together, is running social activities from Wednesday 1st to Saturday 4th. Fun for all ages across the four days. Bring your Picnic blankets and hampers to celebrate the Queens 70 Years on the Throne.

The WI will be on hand to provide refreshments and the Village Hall is kindly […]. A 70 bedroom hotel in Royal Tunbridge Wells, with 14 acres of landscaped grounds. From pm on 5th June The community of Rettendon and the bell ringers at the […]. Starting with light lunches 12 noon till 2pm, then followed by teas from 2pm until 5pm.

There will be no charge for this so bring your own tiaras and robes, partake appropriately and have a good catch-up. Join us at Allington Library on the 31 May, from 2pm for storytime and crafts with a Jubilee theme! A Picnic in the Park on the Recreation ground.

We hope to have an ice cream […]. A picnic in the Park on the Recreation Ground. We have live music from noon-4pm. An ice cream van will be visiting too. The event starts at 7. The beacon will be lit at 9. The event is free […]. Entry is free. There will be collection boxes for various charities. Church Opening Times […]. Please bring your own picnic, there will be limited food to purchase on the day. As our way of celebrating this wonderful occasion you can enjoy a special steam hauled Sparkling Afternoon Tea train at the Spa Valley Railway.

Combine the elegance of steam train travel with a quintessential […]. Images include Royal homes such as […]. Community celebration party honouring Her Majesty, Queen Elizabeth marking the seventieth year of her reign. Group of friends, family and neighbours joining together to celebrate the Queens Platinum Jubilee. Plenty of food and drink with music courtesy of our talented neighbours.

Party games and some history on the interactions neighbours have had with the Queen over the 70 years. Open mic session for those bold on the day and delicious […]. The first time Totternhoe St Giles Bellringers will peal with a full compliment of tollers since the last Jubilee. A free community event with live music, community stalls, tea and cakes, pizza van and ice cream van.

Or bring your own picnic. Thursday 2 June, from 9. Join us as we light our own ceremonial Beacon at The Capitol. The event will be supported by a Bugler, […]. Free event to be held at Cove Cricket Club. Book your place via Eventbrite on our website. Buy a boxed tea or bring your won picnic, biscuit decorating, lucky dip, name the corgi, make your own bunting. Music and cricket. A community day to get together and celebrate.

The Hub Cafe customers love our home made scones, cakes and desserts so what better way to celebrate the Platinum Jubilee? Please Join us at Hawkenbury pavillion Saturday 4th June pm bring a picnic blanket, picnics, bring family and friends for some fun and games There will be ice cream, food and drink vendors alongside activities by local communities and businesses. Rayleigh Town Museum, is a local history museum housed in a 14th Century building in the centre of the town, we have thousands of items and documents of interest and relevance to the town.

To celebrate the jubilee we are holding an exhibition of royal memorabilia, featuring items and photographs of previous local royal visits […]. The organisers provide tables and chairs for people with entertainment and guests provide their own food and drink. Competitions include the best dressed table for six people and the best platinum pudding.

Jubilee picnic including entertainment from crazy beans. Decorate a crown competition with prizes, flags and bubble wands for all whilst stocks last! Bring your gazebo, your gran, scones and jam and have some fun with the BBH clan!

Bring your picnic, blankets and as many friends and family as you can rally to enjoy this once in a lifetime event. Event starts from 2pm and will run until the early evening. The service will be sung by the Parish Choir of Holy Trinity Church Rayleigh in our refurbished church, and accompanied by our new organ. To mark the Jubilee, we have a special display of Coronation and Royal themed memorabilia, with themed activities for younger visitors.

Our dedicated team of volunteers will be on hand to welcome […]. The event will conclude with a Beacon Lighting at 9. I am an individual who has been a Royal family fan for 40 years since the day of Princess Diana and Prince Charles Wedding. I have a huge private collection of memorabilia. It will be […]. Paddock Wood Youth Council will be hosting a picnic in the park on Sunday 5th June from pm at Putlands field with a bar, bouncy castle, pop up skate park and music. Please come along and enjoy!

The days events will include music, cocktails, a penalty shoot-out, sumo wrestling suit competition for Macmillan, picnic box lunches and a take-away menu , Royal line-up selfies and more! The event will run […].

Bring a picnic and your friends and family and take part in some great activities from local community groups. Bring your own picnic, chairs, blanket, table. Live music all afternoon — Traditional games for all the family — croquet, cricket skills, giant jenga and much more. Local beers, lagers, ciders on sale.

Vintage vehicles. Write messages of congratulations to […]. Drop in between Can you find all the Royal objects hidden around the […]. Help make the library bloom for Victoria Day by making a bee, butterfly or flower for us to display. On Sunday 5th June following our Service for Pentecost at 11am we are serving a Buffet picnic lunch on the grass weather permitting We will be lead in service by our Pastor Steve Mayo.

All are welcome to join us. Paint a flowerpot or decorate a crown. Fun for all the family. Free drop in event. Donations welcome. Local acts and stage schools. Stalls, food and community groups will all be attending. Free entry to all! We will be enjoying talks by local Members of Parliament to remember and help celebrate such a wonderful achievement by the Queen.

As well as a local Sussex opera singer will entertain and help in national celebration in […]. With cakes and sandwiches to suit the occasion, our themed tea is one of the tastiest […]. Come along and enjoy a Sparkling Afternoon Tea with live entertainment and join the party at Albert Court. Take a tour of this beautiful development, visit our show suite and meet the vibrant community. Everyone is welcome […]. The Crowborough Locomotive Society was formed in to build, maintain and operate a miniature railway in the grounds of the leisure centre in Crowborough, East Sussex.

It is our aim to provide a unique and fun attraction to the surrounding area which can be enjoyed by all ages whilst encouraging the interest of engineering […]. Local florists and flower clubs will be taking part and members of the church and other organisations will display arts and crafts […]. The venue offers cremation facilities and a Garden of Remembrance for cremated remains in a beautifully-landscaped setting, with manicured gardens and restful country views.

Hullbridge Parish Council will be encouraging residents to attend a Picnic in the Park on the 5th June with family and friends to celebrate the Jubilee. Join us for our family Platinum Jubilee Celebration! There will be a BBQ, bouncy castle, live music and more! All are welcome, but please RSVP via email to hello thevineuk. We are a pet shop in Woodley, Reading and will be celebrating our own 10 year anniversary on the Jubilee weekend. Children are invited to design a corgi crown and will be in with the chance of winning a Percy Pig sweetie hamper.

Southeast Communities Rail Partnership works to connect communities and the railway across Sussex, Kent and Surrey, helping shape the future of the railways and bringing local stations into the heart of the community. Come along to this […]. Fiona […]. The Elstead Jubilee Committee was set up in October to plan celebration events over the extended Bank Holiday weekend of June Below […].

Barton-le-Clay Parish Council has been working with a group of representatives from local groups, church and schools to decorate the village centre Red, White and Blue in celebration of The Queens 70th reign. We are a Charity set up to maintain the Clavering Jubilee Field, which is a space to recreate.

We will be holding various activities on the field on the Saturday including a Big Lunch to bring all villagers together in one place and at the same time w will be hosting various other activities such […]. We are a Day Centre for local elderly residents to socialise, enjoy home cooked food and participate in activities. We will be providing an afternoon tea for approximately 50 of our elderly residents who would otherwise be alone, together with live entertainment.

We will also be arranging for clients who require it, to be transported […]. Memory Lane Cafe is an organisation totally staffed by volunteers supporting carers of people living with the effects of dementia in a social environment. We provide refreshments, entertainment, emotional support and sign-posting to additional services if required. We will be celebrating the Platinum Jubilee by providing an afternoon tea.

Thursday 2nd June Lantern parade, beacon lighting and fireworks Refreshment, drinks and entertainment in Calthorpe Park Friday 3rd June A day of sports including a cycle ride, football, tennis, athletics, archery, […]. Two performances of music, sketches, poetry and monologues of Kings and Queens through the ages.. One at 2. U3A Recorder Consort to be included. Some tribute artists look like their idol, some sound like their idol.

Paul Bacon as Elton John does both! Meeting on the village Green to celebrate the Jubilee. Bring your own food and drink, there will be a barbecue for cooking on. Tables, Chairs, Music and Games all provided. Open to everyone to attend. The renowned Vasari Singers is regarded as one of the leading chamber choirs in Britain. Under the direction of its founder-conductor Jeremy Backhouse, Vasari Singers performs a wide range of repertoire from Renaissance to contemporary with a special Platinum Jubilee flavour.

There will be a Service of Commemoration and Thanksgiving for the seventy years of her reign at The service will include well-known hymns, […]. Themed Village Fete around the Platinum Jubilee with food stalls, craft stalls, entertainments including arena events. Between 3pm […]. All of this accompanied by face painting and fun activities […]. Tuck into your royal delights […]. Fairwarp lies within Ashdown Forest and the by-laws forbid naked flames on the village green so instead we are going to dress in fairy lights.

Join us on the village green from 7. The Foresters Arms will be open to provide any […]. We will a long table and chairs on tThe Street for residents to join together for a picnic lunch. Bring your own lunch or book a picnic lunch from The Foresters Arms and, of course, the pub will be there to provide us with drinks during the day. Please join us at our fete to celebrate the Queens Platinum Jubilee. Put your hat on and come to the village hall for a fun-filled afternoon tea. Watch the Derby live on our big screen and take part in our free fun sweepstake.

Menu Glass of Prosecco A selection of sandwiches Scones, jam and cream A selection of cakes and sweetmeats Tea or coffee Special prize for […]. The exhibition will be available to be viewed as follows: Thursday 2 June — We are hosting three events in the run up to the Jubilee. Village organisations and musicians providing food and entertainment during the afternoon for the community.

BBQ, sweet and savoury foods, range of music throughout afternoon and into evening, sporting activities, stalls and sideshows, prize for best decorated Jubilee Bonnet. Starting […]. Stotfold Bowls Club is open in the summer months for members of all ages to play bowls. Just turn up and have a go at target bowls and other games.

Please make sure you wear […]. In The Victory Hall was bequeathed to villagers. Run as a charity trust and run maintained by locals in their own time. The Farley Hill Village Society raises money through events and donations in this tiny hamlet in berksire and are delighted to celebrate our Queens jubilee on the 5th June with BBQ, games, […]. There will be classic pieces of Coronation and National music and singing at pm on Thursday 2nd June, prior to the Beacon lighting at pm.

On 2nd June, a specially commissioned beacon made by a local crafts person will be lit on Plaistow village green. The event begins at and includes complementary refreshments and patriotic […]. Don any outfit you fancy and join the procession which leaves the […].

We have Hungary v England on the big screen at 5pm followed by a disco and karaoke with bbq in the evening. Close out the Platinum Jubilee with local drag monarchs and a royally-themed bingo!! Platinum Play A joint Christian service at St. A special extended bank holiday weekend at the beginning of June will provide an opportunity for communities and people […].

Have you no street party to go to? Then at 2. We are a local restaurant located on the A1 South between Biggleswade and Baldock. On Saturday we will have fun full of activities for all the family to celebrate the Queen Jubilee, in our restaurants including indoor and outdoor activities. This event is open to any local Hartley Wintney residents […]. The Plough Inn is a relaxed and unpretentious freehold family run pub, bar and restaurant with its very own large patio garden managed by Becky, Adam and Debbie.

The Plough is centrally located in the heart of the picturesque villages of Sutton Valence, Langley and only 2 miles away from the magnificent Leeds Castle. June 2 — Leeds Village will be gaining their own permanent beacon — located at the front […]. Hartley Wintney Parish Council will lead a candle lit procession from the Hartley Wintney Football Club to the Jubilee Fields where there will be a short ceremony and the Jubilee Beacon will be lit, along with beacons all over the country.

Please give us your idea of what you think the carriage would look like either inside, outside, or both! There […]. Maresfield village lunch club in conjunction with maresfield parish council are organizing an outdoor picnic venue at maresfield recreation ground. Debden Parish Council, in collaboration with other village community groups, has organised events over a four day period to celebrate the Queens Platinum Jubilee for the weekend starting 2nd June Leeds Parish Council, will be gaining their permanent beacon, crafted by a local blacksmith, with a pole donated locally.

Leeds Village will call on all our Village to gather and join as we light the beacon for the first time, in front of St Nicholas Church, and join in the Lighting of the Beacons around […]. Reminiscent arts and crafts festival for elderly people in Reading and nearby areas. A celebration of the creativity of active ageing minds, treasured memories, and the day-to-day lives of older people throughout the reign of HRH The Queen. The arts and crafts sessions will be focused on creating beautiful items related to the Jubilee which […].

The Picnic in the Park is the centrepiece of the various events being held in the villages of Flitton, Greenfield and Wardhedges. Lots of antique and vintage collectable commemorative wear. Tea Room with a Royal theme including Victoria sponge cake.

As we celebrate the reign of Her Majesty Queen Elizabeth II, we are taking a look back at all the powerful, strong and cultured queens in our history. Parish Council 2. We will be hosting a beacon lighting on 2nd June at the St. Johns recreation ground in the village bar open from 6. Then on 3rd June we have the fete and dog show in the War Memorial recreation Ground from 2.

Kangaroos Mid Sussex is a charity which support children and adults with learning disabilities and their families. We provide different fun activities for our members to enable them socialise with their peers, get out and about in their community and develop their independence, confidence and self-esteem.

Our Picnic in the Park will enable our members […]. It was introduced in to help deliver local initiatives beneficial to the business community. The trail will have younger children 7 and under find the crowns in the window shops and older children 8—11 will need to read the royal fact in each window and write […].

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There will be many events and activities celebrating the Jubilee across the UK Commonwealth and beyond in the months leading up to the special extended bank holiday weekend from Thursday 2 to Sunday 5 June

Alina smith free beer torrent Two performances of music, sketches, poetry and monologues of Kings and Queens through the ages. Broxbourne Borough Council is marking the Platinum […]. Thanksgiving service for the Queen 7. Click we show that mouse pronuclear transfer can be used to ameliorate such reprogramming defects. We will be celebrating The Queens Platinum Jubilee with a good old-fashioned knees up and singalong.
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