Protein phosphorylation and cell diversification in the mouse early embryo.

This dissertation reports the results of studies into the control of compaction of the mouse preimplantation embryo. Compaction is a post-translationally controlled rearrangement of cell contacts and the cytoskeleton that occurs at the 8-cell stage of development. This re-arrangement seems to be necessary for the differentiation of the two cell types present in the blastocyst. Protein phosphorylation is a post-translational modification believed to be important in the modulation of cell shape and cytoskeletal assembly. It is therefore feasible to propose a role for protein phosphorylation in compaction. Two types of approach have been used to investigate the possible role of protein phosphorylation in compaction. Firstly, embryos have been treated with two drugs, 6-dimethylaminopurine (DMAP) and a phorbol ester (phorbol myristate acetate, PMA), each of which seems to affect both protein phosphorylation and compaction. DMAP is an adenine analogue and putative inhibitor of protein phosphorylation that was found to perturb the cell cycle of mouse embryos. In addition, DMAP caused rapid cellular flattening of 4-cell and 8-cell embryos. However, this flattening was not accompanied by cell polarisation and did not seem to be mediated by the cell adhesion molecule uvomorulin. It is therefore unlikely to be related directly to the flattening that occurs at compaction. Phorbol esters, such as PMA, are potent stimulators of the membrane-associated, Ca2+- and phospholipid-dependent protein kinase, protein kinase C (PKC). Incubation in medium containing PMA had some effects on the cytoskeleton of oocytes and early embryos but caused severe, widespread disassembly of the cytoskeleton and reversal of flattening in 8-cell embryos. These effects of PMA, seen specifically at the 8-cell stage, may be related to the spatially restricted disassembly of the cytoskeleton that occurs naturally during compaction at the 8-cell stage. This interpretation provides indirect evidence for a possible role for PKC activity, and hence protein phosphorylation, in the process of compaction. The relationship between protein phosphorylation and the events occurring at the 8-cell stage has been examined more directly by labelling 4-cell and 8-cell embryos with [32P]orthophosphate and examining the phosphoproteins obtained by one and two-dimensional gel electrophoresis. By synchronising groups of embryos precisely to successive cleavage divisions prior to labelling, changes in phosphoprotein profile associated with passage through the 4-cell and 8-cell stages have been described. While many of the 32P-labelled phosphoproteins detectable after electrophoresis in one or two dimensions are similar at each stage examined, there are some changes associated specifically with passage through the 8-cell stage which may be related to the cell flattening and polarisation occurring at this time. In addition, the profile of 8-cell embryos differed according to the duration of pulse-labelling with [32Pjorthophosphate or the inclusion of "chase" periods. Finally, several treatments that affect features of compaction, including exposure to DMAP and PMA, have been used to assess the link between the observed changes in phosphoprotein profile and the events of compaction. Embryos were also incubated in protein synthesis inhibitors, which cause premature cell flattening in 4-cell embryos and in Ca2+-free medium, which prevents intercellular flattening and delays polarisation of 8-cell blastomeres. In each case, the relative labelling intensity of some of the phosphoproteins characteristic of untreated 8-cell embryos was altered. The behaviour of these phosphoproteins suggests that they may be important in the mechanism by which cells flatten and polarise or in the maintenance of flattened, polarised, cells; they now provide a focus for future study

Using peer review to support development of community resources for research data management

This work is licensed under a Creative Commons 1.0 Public Domain Dedication. The definitive version was published in Journal of eScience Librarianship 6 (2017): e1114, doi:10.7191/jeslib.2017.1114.To ensure that resources designed to teach skills and best practices for scientific research data sharing and management are useful, the maintainers of those materials need to evaluate and update them to ensure their accuracy, currency, and quality. This paper advances the use and process of outside peer review for community resources in addressing ongoing accuracy, quality, and currency issues. It further describes the next step of moving the updated materials to an online collaborative community platform for future iterative review in order to build upon mechanisms for open science, ongoing iteration, participation, and transparent community engagement.DataONE is supported by US National Science Foundation Awards 08- 30944 and 14-30508, William Michener, Principal Investigator; Matthew Jones, Patricia Cruse, David Vieglais, and Suzanne Allard, Co-Principal Investigators

The Effect of Dietary Tartrazine on Brain Dopamine and the Behavioral Symptoms of Attention Deficit Hyperactivity Disorder

Attention Deficit Hyperactivity Disorder is a neurodevelopmental disorder correlated with a decrease in brain dopamine and an increase in behavioral symptoms of hyperactivity and impulsivity. This experiment explored how tartrazine (Yellow #5) impacts these symptoms. After tartrazine administration to Spontaneously Hypertensive Rats (SHR), dopamine concentrations in regions of brain tissue were measured using Enzyme-Linked Immunosorbent Assay analysis. Behavioral testing with a T-maze and open field test measured impulsivity and hyperactivity, respectively. Results indicate that dietary tartrazine increases hyperactive behaviors in the SHR. However, results do not indicate a relationship between dietary tartrazine and brain dopamine. No conclusions regarding the relationship between dietary tartrazine and impulsivity were drawn

Prospectus, September 21, 1983

CONSTRUCTION PROGRESSES; Manager runs unique Hardee\u27s; Elect your senator Sept. 21-22; Prisons pursue harsher sentencing; Student admits scary reality; Mr. Roberts: never a water shortage; StuGo elections held this week; Paxton- town of peace ; Working women ignored in many countries; Rape affects into past; Pregnancy problem for teenagers; Dress for success; Segregation exists-- progress is little; Classified; Stu-Go happenings; Produce prices will be stable; Rockabilly E.B.\u27s are Movin\u27 Up ; Campus Paperback Bestsellers; New & Recommended; Ballet Nacional Español to be at Assembly Hall; Stray Cats definitely strut; Crenshaw\u27s Field Day; UI Ticket Exchange Area designated; Gym guidelines; Henkels looks to volleyball nationals; Fast Freddy Contest; Traffic Changes for UI footballhttps://spark.parkland.edu/prospectus_1983/1010/thumbnail.jp

CropPol: a dynamic, open and global database on crop pollination

Seventy five percent of the world's food crops benefit from insect pollination. Hence, there has been increased interest in how global change drivers impact this critical ecosystem service. Because standardized data on crop pollination are rarely available, we are limited in our capacity to understand the variation in pollination benefits to crop yield, as well as to anticipate changes in this service, develop predictions, and inform management actions. Here, we present CropPol, a dynamic, open and global database on crop pollination. It contains measurements recorded from 202 crop studies, covering 3,394 field observations, 2,552 yield measurements (i.e. berry weight, number of fruits and kg per hectare, among others), and 47,752 insect records from 48 commercial crops distributed around the globe. CropPol comprises 32 of the 87 leading global crops and commodities that are pollinator dependent. Malus domestica is the most represented crop (32 studies), followed by Brassica napus (22 studies), Vaccinium corymbosum (13 studies), and Citrullus lanatus (12 studies). The most abundant pollinator guilds recorded are honey bees (34.22% counts), bumblebees (19.19%), flies other than Syrphidae and Bombyliidae (13.18%), other wild bees (13.13%), beetles (10.97%), Syrphidae (4.87%), and Bombyliidae (0.05%). Locations comprise 34 countries distributed among Europe (76 studies), Northern America (60), Latin America and the Caribbean (29), Asia (20), Oceania (10), and Africa (7). Sampling spans three decades and is concentrated on 2001-05 (21 studies), 2006-10 (40), 2011-15 (88), and 2016-20 (50). This is the most comprehensive open global data set on measurements of crop flower visitors, crop pollinators and pollination to date, and we encourage researchers to add more datasets to this database in the future. This data set is released for non-commercial use only. Credits should be given to this paper (i.e., proper citation), and the products generated with this database should be shared under the same license terms (CC BY-NC-SA). This article is protected by copyright. All rights reserved

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Guidelines for publishing papers containing theory and modeling

The use of quantitative approaches in cell biology, including modeling and theory, has been increasing steadily in the past decade. A number of seminal works that combine experiments and modeling have made a significant impact on the field (see Hill and Kirschner, 1982; Peskin et al., 1993; Barkai and Leibler, 1997, to name but a few). Mathematical modeling can help when our intuition fails or misleads us. It can provide a precise language for understanding complex cell biological phenomena. Modeling represents a new addition to our repertoire of strategies to understand biological systems

Effects of a combined protein and antioxidant supplement on recovery of muscle function and soreness following eccentric exercise

Abstract Background An acute bout of eccentric contractions (ECC) cause muscle fiber damage, inflammation, impaired muscle function (MF) and muscle soreness (MS). Individually, protein (PRO) and antioxidant (AO) supplementation may improve some aspects of recovery from ECC, though have yet to be combined. We sought to determine if combined PRO and AO supplementation (PRO + AO) improves MS and MF following damaging ECC over PRO alone. Methods Sixty sedentary college-aged males participated in a randomized, single–blind, parallel design study of peak isometric torque (PIMT), peak isokinetic torque (PIKT), thigh circumference (TC), and muscle soreness (MS) of knee extensor muscles measured at baseline, immediately after and 1, 2, 6, and 24 h after completion of 100 maximal ECC. Immediately, 6 h, and 22 h post-ECC, participants consumed either: carbohydrate control (CHO; n = 14), PRO (n = 16), or PRO + AO (n = 17). Results At baseline MS, TC, MF, macro- and micro-nutrient intakes, and total work during the ECC were not different between groups (p > 0.05). PIMT and PIKT (both −25%∆), TC (~1%∆) and MS (~35%∆) all changed with time (p  CHO, p  PRO & CHO, p < 0.05). Conclusions Our results suggest PRO facilitates recovery of muscle function within 24 h following ECC, and addition of AO ameliorates MS more than PRO or CHO alone