The Iowa Homemaker vol.6, no.10

Table of Contents V-E-I-S-H-E-A by Gladys Parker, page 1 Is the Home Economics Graduate the Best Homemaker?, page 2 The Season Suggests a Sewing Machine by Cleo Fitzsimmons, page 3 How Shall We Keep Our Food Cold? by Helen Bishop, page 4 4-H Clubs, page 6 Iowa State Home Economics Association Page, page 8 Editorial, page 9 Who’s There and Where, page 10 Research in Canning by Betty Barker, page 11 Vodvil Puts $350 in MacKay Loan Fund by Marcella Alton, page 11 Art Students Compete by Jane Rhoads, page 11 New Meat Course by Helen Goeppinger, page 11 Eternal Question, page 12 Movement to End Waste in Textile Industry, page 13 Analyzes 100 Brands of Sheeting by Mary Moser, page 1

Expression of p16 Within Myenteric Neurons of the Aged Colon: A Potential Marker of Declining Function.

Human colonic neuromuscular functions decline among the elderly. The aim was to explore the involvement of senescence. A preliminary PCR study looked for age-dependent differences in expression of CDKN1A (encoding the senescence-related p21 protein) and CDKN2A (encoding p16 and p14) in human ascending and descending colon (without mucosa) from 39 (approximately 50: 50 male: female) adult (aged 27-60 years) and elderly donors (70-89 years). Other genes from different aging pathways (e.g., inflammation, oxidative stress, autophagy) and cell-types (e.g., neurons, neuron axonal transport) were also examined. Unlike CDKN1A, CDKN2A (using primers for p16 and p14 but not when using p14-specific primers) was upregulated in both regions of colon. Compared with the number of genes appearing to upregulate in association with temporal age, more genes positively associated with increased CDKN2A expression (respectively, 16 and five of 44 genes studied for ascending and descending colon). Confirmation of increased expression of CDKN2A was sought by immunostaining for p16 in the myenteric plexus of colon from 52 patients, using a semi-automated software protocol. The results showed increased staining not within the glial cells (S100 stained), but in the cytoplasm of myenteric nerve cell bodies (MAP2 stained, with identified nucleus) of ascending, but not descending colon of the elderly, and not in the cell nucleus of either region or age group (5,710 neurons analyzed: n = 12-14 for each group). It was concluded that increased p16 staining within the cytoplasm of myenteric nerve cell bodies of elderly ascending (but not descending) colon, suggests a region-dependent, post-mitotic cellular senescence-like activity, perhaps involved with aging of enteric neurons within the colon

A CellAgeClock for expedited discovery of anti-ageing compounds

We aim to improve anti-ageing drug discovery, currently achieved through laborious and lengthy longevity analysis. Recent studies demonstrated that the most accurate molecular method to measure human age is based on CpG methylation profiles, as exemplified by several epigenetics clocks that can accurately predict an individual’s age. Here, we developed CellAgeClock, a new epigenetic clock that measures subtle ageing changes in primary human cells in vitro . As such, it provides a unique tool to measure effects of relatively short pharmacological treatments on ageing. We validated the CellAgeClock against known longevity drugs such as rapamycin and trametinib. Moreover, we uncovered novel anti-ageing drugs, torin2 and Dactolisib (BEZ-235), demonstrating the value of our approach as a screening and discovery platform for anti-ageing strategies. The CellAgeClock outperforms other epigenetic clocks in measuring subtle ageing changes in primary human cells in culture. The tested drug treatments reduced senescence and other ageing markers, further consolidating our approach as a screening platform. Finally, we show that the novel anti-ageing drugs we uncovered in vitro , indeed increased longevity in vivo . Our method expands the scope of CpG methylation profiling from measuring human chronological and biological age from human samples in years, to accurately and rapidly detecting anti-ageing potential of drugs using human cells in vitro , providing a novel accelerated discovery platform to test sought after geroprotectors

A whole genome screen for HIV restriction factors

RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are.Abstract Background Upon cellular entry retroviruses must avoid innate restriction factors produced by the host cell. For human immunodeficiency virus (HIV) human restriction factors, APOBEC3 (apolipoprotein-B-mRNA-editing-enzyme), p21 and tetherin are well characterised. Results To identify intrinsic resistance factors to HIV-1 replication we screened 19,121 human genes and identified 114 factors with significant inhibition of infection. Those with a known function are involved in a broad spectrum of cellular processes including receptor signalling, vesicle trafficking, transcription, apoptosis, cross-nuclear membrane transport, meiosis, DNA damage repair, ubiquitination and RNA processing. We focused on the PAF1 complex which has been previously implicated in gene transcription, cell cycle control and mRNA surveillance. Knockdown of all members of the PAF1 family of proteins enhanced HIV-1 reverse transcription and integration of provirus. Over-expression of PAF1 in host cells renders them refractory to HIV-1. Simian Immunodeficiency Viruses and HIV-2 are also restricted in PAF1 expressing cells. PAF1 is expressed in primary monocytes, macrophages and T-lymphocytes and we demonstrate strong activity in MonoMac1, a monocyte cell line. Conclusions We propose that the PAF1c establishes an anti-viral state to prevent infection by incoming retroviruses. This previously unrecognised mechanism of restriction could have implications for invasion of cells by any pathogen.Published versio

Low Stress Ion Conductance Microscopy of Sub-Cellular Stiffness.

Directly examining subcellular mechanics whilst avoiding excessive strain of a live cell requires the precise control of light stress on very small areas, which is fundamentally difficult. Here we use a glass nanopipet out of contact with the plasma membrane to both exert the stress on the cell and also accurately monitor cellular compression. This allows the mapping of cell stiffness at a lateral resolution finer than 100 nm. We calculate the stress a nanopipet exerts on a cell as the sum of the intrinsic pressure between the tip face and the plasma membrane plus its direct pressure on any glycocalyx, both evaluated from the gap size in terms of the ion current decrease. A survey of cell types confirms that an intracellular pressure of approximately 120 Pa begins to detach the plasma membrane from the cytoskeleton and reveals that the first 0.66 ± 0.09 μm of compression of a neuron cell body is much softer than previous methods have been able to detect.Biotechnology and Biological Sciences Research Council (Grant ID: BB/L006227/1), Engineering and Physical Sciences Research Council (Grant ID: EP/H01098X/1), Medical Research Council (Grant IDs: G0701057, MR/K501372/1), Herchel Smith Postdoctoral Fellowship, Royal Society of Chemistry Analytical Chemistry Trust Fun

Cellular Senescence: Defining a Path Forward.

Cellular senescence is a cell state implicated in various physiological processes and a wide spectrum of age-related diseases. Recently, interest in therapeutically targeting senescence to improve healthy aging and age-related disease, otherwise known as senotherapy, has been growing rapidly. Thus, the accurate detection of senescent cells, especially in vivo, is essential. Here, we present a consensus from the International Cell Senescence Association (ICSA), defining and discussing key cellular and molecular features of senescence and offering recommendations on how to use them as biomarkers. We also present a resource tool to facilitate the identification of genes linked with senescence, SeneQuest (available at http://Senequest.net). Lastly, we propose an algorithm to accurately assess and quantify senescence, both in cultured cells and in vivo

An investigation of the small polypeptides of photosystem II

Photosystem II catalyses the light-driven electron transfer from water to plastoquinone. At present the complex is thought to contain at least 25 different polypeptides. In eukaryotes the components of PSII are encoded for both chloroplast and nuclear genes. Whilst information on the structure of the PSII complex is beginning to emerge, the role that the majority of the subunits play remains unknown. This thesis investigates three of these subunits, the nuclear-encoded subunit PSII-W, the chloroplast-encoded subunit PSII-M and PSII-Z (previously the hypothetical chloroplast protein Ycf9). The gene that encodes the PSII-W protein has been cloned and sequenced from the photosynthetic alga Chlamydomonas reinhardtii. Analysis of the derived amino acid sequence has enabled the characterisation of the PSII-W protein. Antibodies to the mature form of the protein have been raised and used to perform initial expression analysis for the protein and to examine claims that the protein may also be present in a second thylakoid membrane complex, photosystem I. We establish that the PSII-W protein is located exclusively in the PSII complex of C. reinhardtii. Further work has been undertaken to clone and sequence the psbW gene from the moss Physcomitrella patens. Disruption and deletion mutants in the psbZ (previously ycf9) and psbM genes of the cyanobacterium Synechocystis sp. PCC 6803 respectively, have been created by using the kanamycin resistance cassette as a selectable marker. Analysis of these mutants has been undertaken in an attempt to determine the role that each of the proteins plays in the PSII complex. Whilst no definitive role for either of the subunits is established a number of possible functions are ruled out

Current Understanding of the Role of Senescent Melanocytes in Skin Ageing

: Melanocytes reside within the basal epidermis of human skin, and function to protect the skin from ultraviolet light through the production of melanin. Prolonged exposure of the skin to UV light can induce irreparable DNA damage and drive cells into senescence, a sustained cell cycle arrest that prevents the propagation of this damage. Senescent cells can also be detrimental and contribute to skin ageing phenotypes through their senescence-associated secretory phenotype. Senescent cells can act in both an autocrine and paracrine manner to produce widespread tissue inflammation and skin ageing. Recently, melanocytes have been identified as the main senescent cell population within the epidermis and have been linked to a variety of skin ageing phenotypes, such as epidermal thinning and the presence of wrinkles. However, the literature surrounding melanocyte senescence is limited and tends to focus on the role of senescence in the prevention of melanoma. Therefore, this review aims to explore the current understanding of the contribution of senescent melanocytes to human skin ageing

Role for Centromeric Heterochromatin and PML Nuclear Bodies in the Cellular Response to Foreign DNA

Nuclear spatial positioning plays an important role in the epigenetic regulation of eukaryotic gene expression. Here we show a role for nuclear spatial positioning in regulating episomal transgenes that are delivered by virus-like particles (VLPs). VLPs mediate the delivery of plasmid DNA (pDNA) to cell nuclei but lack viral factors involved in initiating and regulating transcription. By tracking single fluorescently labeled VLPs, coupled with luciferase reporter gene assays, we found that VLPs transported pDNA to cell nuclei efficiently but transgenes were immediately silenced by the cell. An investigation of the nuclear location of fluorescent VLPs revealed that the pDNAs were positioned next to centromeric heterochromatin. The activation of transcription by providing viral factors or inhibiting histone deacetylase activity resulted in the localization to euchromatin regions. Further, the activation of transcription induced the recruitment of PML nuclear bodies (PML-NBs) to the VLPs. This association did not play a role in regulating transgene expression, but PML protein was necessary for the inhibition of transgene expression with alpha interferon (IFN-α). These results support a model whereby cells can prevent foreign gene expression at two levels: by positioning transgenes next to centromeric heterochromatin or, if that is overcome, via the type I IFN response facilitated by PML-NB recruitment

The Iowa Homemaker vol.6, no.10

Table of Contents V-E-I-S-H-E-A by Gladys Parker, page 1 Is the Home Economics Graduate the Best Homemaker?, page 2 The Season Suggests a Sewing Machine by Cleo Fitzsimmons, page 3 How Shall We Keep Our Food Cold? by Helen Bishop, page 4 4-H Clubs, page 6 Iowa State Home Economics Association Page, page 8 Editorial, page 9 Who’s There and Where, page 10 Research in Canning by Betty Barker, page 11 Vodvil Puts $350 in MacKay Loan Fund by Marcella Alton, page 11 Art Students Compete by Jane Rhoads, page 11 New Meat Course by Helen Goeppinger, page 11 Eternal Question, page 12 Movement to End Waste in Textile Industry, page 13 Analyzes 100 Brands of Sheeting by Mary Moser, page 16</p