Proximity of IQOS and JUUL points of sale to schools in Israel: a geospatial analysis

SIGNIFICANCE: Exploring proximity and density of heated tobacco product (HTP) and electronic nicotine delivery system (ENDS) points of sale (POS) to schools is critical for understanding youth marketing exposure and informing policy and enforcement to protect youth. This study examined IQOS and JUUL POS (prominent HTPs and ENDS), specifically their proximity to and density around schools in Israel. METHODS: Using geospatial analysis and IQOS/JUUL website data, distance matrices were used to calculate distance from each school in Israel (grades 1-12) to the nearest POS and number of POS within 1 km, accounting for schools\u27 neighbourhood socioeconomic status (SES) ranking. RESULTS: An average of 8.7 IQOS POS and 5.2 JUUL POS were within walking distance (1 km) from schools. Average distances from schools to nearest IQOS and JUUL POS were 954 m (median=365 m) and 1535 m (median=579 m), respectively. The percentages of schools with at least one IQOS or JUUL POS within 1 km were 86% and 74%, respectively. The average numbers of POS within 1 km of schools in low-SES, middle-SES, and high-SES neighbourhoods were 7.5, 9.9, and 7.6 for IQOS and 4.1, 5.9, and 5.5 for JUUL, respectively. Median distances from schools in low-SES, middle-SES, and high-SES neighbourhoods to nearest POS were 428 m, 325 m, and 403 m for IQOS and 1044 m, 483 m, and 525 m for JUUL. CONCLUSIONS: Youth experience high environmental exposure to IQOS and JUUL POS, particularly IQOS. POS were more densely located near schools in middle-SES neighbourhoods. Thus, regulating HTP and ENDS POS near schools and in certain neighbourhoods is key to reducing youth population impact in Israel and elsewhere

A Systematic Approach to Pair Secretory Cargo Receptors with Their Cargo Suggests a Mechanism for Cargo Selection by Erv14

<div><p>The endoplasmic reticulum (ER) is the site of synthesis of secreted and membrane proteins. To exit the ER, proteins are packaged into COPII vesicles through direct interaction with the COPII coat or aided by specific cargo receptors. Despite the fundamental role of such cargo receptors in protein traffic, only a few have been identified; their cargo spectrum is unknown and the signals they recognize remain poorly understood. We present here an approach we term “PAIRS” (pairing analysis of cargo receptors), which combines systematic genetic manipulations of yeast with automated microscopy screening, to map the spectrum of cargo for a known receptor or to uncover a novel receptor for a particular cargo. Using PAIRS we followed the fate of ∼150 cargos on the background of mutations in nine putative cargo receptors and identified novel cargo for most of these receptors. Deletion of the Erv14 cargo receptor affected the widest range of cargo. Erv14 substrates have a wide array of functions and structures; however, they are all membrane-spanning proteins of the late secretory pathway or plasma membrane. Proteins residing in these organelles have longer transmembrane domains (TMDs). Detailed examination of one cargo supported the hypothesis that Erv14 dependency reflects the length rather than the sequence of the TMD. The PAIRS approach allowed us to uncover new cargo for known cargo receptors and to obtain an unbiased look at specificity in cargo selection. Obtaining the spectrum of cargo for a cargo receptor allows a novel perspective on its mode of action. The rules that appear to guide Erv14 substrate recognition suggest that sorting of membrane proteins at multiple points in the secretory pathway could depend on the physical properties of TMDs. Such a mechanism would allow diverse proteins to utilize a few receptors without the constraints of evolving location-specific sorting motifs.</p> </div

Assessment of IQOS Marketing Strategies at Points-of-Sale in Israel at a Time of Regulatory Transition

Introduction: IQOS, a tobacco heating system, and accompanying tobacco sticks (HEETS) entered the Israeli market in 2016, prior to rapid regulatory change. This study assessed IQOS marketing strategies and regulatory compliance at IQOS and/or HEETS point-of-sale (POS) in Israel in December 17, 2019 to January 7, 2020, after the ban on advertisement went into effect in March 8, 2019. Aims and Methods: Research staff audited 80 randomly selected IQOS and/or HEETS POS in four cities using a structured form to assess store types, product placement, price, promotional strategies, and regulatory compliance. POS data were linked to neighborhood characteristics, including socioeconomic status, ethnicity, and proximity (under 300 m) to schools. Results: Almost half of the stores (48.7%) were convenience stores. HEETS were visible to the customers in 46.1% of POS, 35% carried at least four HEETS colors, 20.0% had IQOS and/or HEETS special displays, and 13.8% displayed HEETS near youth-oriented merchandise. Mean HEETS pack price was US $8.7 (range: US$7.5-11.3), 27% more than the least expensive cigarette pack, and 39% less than the most expensive cigarette. HEETS promotions were uncommon. Compliance with the newly introduced advertisement ban was fairly high for HEETS (94.8%). Only one POS was located in a low-socioeconomic status area; 68.7% were near a school. Conclusions: The relatively limited IQOS and/or HEETS marketing at POS suggests that, with regulatory changes, online or other forms of marketing might be prioritized. IQOS may be promoted to higher socioeconomic status populations, as indicated by pricing and POS neighborhood characteristics. Access near schools and placement near youth-oriented merchandise are potential concerns necessitating further research. Implications: Globally, the POS is considered the least regulated channel for advertising and marketing of tobacco products. Assessing IQOS marketing strategies at the POS provides valuable findings that can inform regulatory efforts in Israel and other countries as well. Limited IQOS and/or HEETS marketing at POS suggests that primary marketing strategies may shift to online or other channels as regulatory contexts become more progressive and/or restrictive. Ongoing surveillance of IQOS via online marketing and POSs, specifically with regard to product placement and proximity to schools, is needed

The cytoplasmic tail of Sys1 accelerates the ER exit of short TMD variants of Mid2.

<p>(A) The C-terminal cytoplasmic tail of the polytopic membrane protein Sys1. The end of the last of its four TMDs is shown, along with the DXE motif that binds COPII <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001329#pbio.1001329-Votsmeier1" target="_blank">[38]</a>. Also shown is the structure of the Mid2 polyleucine variants with the Mid2 tail replaced with that of Sys1 (see also <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001329#pbio-1001329-g004" target="_blank">Figure 4B</a>). (B) Anti-GFP immunoblots of whole cell lysates from yeast expressing the indicated polyleucine TMD variants of Mid2-GFP with tail replaced with that of Sys1. The variants were expressed under the control of the <i>GAL1</i> promoter from constructs integrated at the MID2 locus. The cells were induced with galactose for 2 h, and then harvested at the times indicated after replacing the medium with that containing 2% glucose. The arrows indicated the ER form (ER) and Golgi modified form (G) of Mid2-GFP, and free GFP. For reasons that are unclear we could not obtain cells expressing the L26 variant at the same levels as the other constructs and so it was omitted. The results are representative of three independent experiments. (C) As Mid2L18S-GFP in (B), except that the DXE motif is mutated to AXE. (D and E) Graphs of ER exit of the constructs shown in (B and C), as calculated for <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001329#pbio-1001329-g004" target="_blank">Figure 4F</a>.</p

PAIRS: a systematic approach for identification of cargo for a given cargo receptor.

<p>Libraries containing deletion in trafficking-related proteins or secretory pathway proteins tagged with GFP were assembled. Using the SGA methodology they were crossed against query strains carrying either a cargo tagged with GFP or a deletion in one of nine different cargo receptors with the assistance of automated cell manipulation techniques. Libraries were then automatically imaged for GFP localization followed by manual inspection of the images, enabling detection of ER retention due to a given deletion.</p

PAIRS allows pairing of secretory pathway cargo with their respective cargo receptors.

<p>Mutations in nine cargo receptors were studied for their effect on cargo retention. Shown are the proteins that displayed ER retention phenotype on the background of (A) mutations in Δ<i>erv29</i>, (B) Δ<i>gsf2</i>, (C) Δ<i>erv26</i>, (D) Δ<i>emp47</i>, (E) <i>shr3-DAmP</i> relative to a control (wild-type [WT]) strain during logarithmic growth. Shown for each is a cartoon hypothesizing the common denominator of its respective cargo. (F) Deletion of Erv14 caused widespread ER retention. Shown are localizations of eight GFP-tagged proteins during logarithmic growth in control strains (WT) relative to Δ<i>erv14.</i> For a full list, see <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001329#pbio.1001329.s002" target="_blank">Figure S2A</a>. Pictures were taken at a magnification of 60×.</p

TMD length dependent localization to the trans Golgi can be independent of endocytosis or differential ER exit rate.

<p>(A) ER exit rates of the Mid2-GFP with either 16 or 24 leucines in the TMD and the Sys1 cytoplasmic tail in wild-type or a strain lacking Erv14. Cells were induced with galactose and chased as in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001329#pbio-1001329-g005" target="_blank">Figure 5</a>. Removal of Erv14 results in there being no difference in the ER exit rate of the 16 and 24 leucine TMD variants. (B) Wide-field fluorescent micrographs of live yeast cells expressing Mid2-GFP with either 16 or 24 leucines in the TMD and the Sys1 cytoplasmic tail. The chimeras are constitutively expressed from the <i>MID2</i> promoter by integration at the endogenous <i>MID2</i> locus in strain lacking Erv14. Despite similar ER exit rates the two chimeras have a different intracellular distribution. Scale bars 1 µm.</p