The obligate intracellular bacterium Chlamydia elicits a great burden on global public health. C. trachomatis is the leading bacterial cause of sexually transmitted infection and also the primary cause of preventable blindness in the world. An essential determinant for successful infection of host cells by Chlamydia is the bacterium's ability to manipulate host cell signaling from within a novel, vacuolar compartment called the inclusion. From within the inclusion, Chlamydia acquire nutrients required for their 2-3 day developmental growth, and they additionally secrete a panel of effector proteins onto the cytosolic face of the vacuole membrane and into the host cytosol. Gaps in our understanding of Chlamydia biology, however, present significant challenges for visualizing and analyzing this intracellular compartment. Recently, a reverse-imaging strategy for visualizing the inclusion using GFP expressing host cells was described. This approach rationally exploits the intrinsic impermeability of the inclusion membrane to large molecules such as GFP. In this work, we describe how GFP-or mCherry-expressing host cells are generated for subsequent visualization of chlamydial inclusions. Furthermore, this method is shown to effectively substitute for costly antibody-based enumeration methods, can be used in tandem with other fluorescent labels, such as GFP-expressing Chlamydia, and can be exploited to derive key quantitative data about inclusion membrane growth from a range of Chlamydia species and strains. Video LinkThe video component of this article can be found at

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PurposeBreast cancer is the second leading cause of female cancer mortality in the United States and breast cancer mortality in Asian Americans (AA) is rising by 1.5% per year. However, aggregated AA breast cancer death rates may mask important mortality differences in major AA groups.Population & Setting11,388 AA and 473,927 non-Hispanic White (NHW) females based on the United States Centers for Disease Control and Prevention National Vital Statistics System database 2003-2017.MethodsAge-adjusted mortality rates (AAMR) were used to estimate trends in breast cancer mortality in Asian Indians, Chinese, Filipinas, Japanese, Koreans, Vietnamese, and non-Hispanic Whites from 2003–2017, with attention to annual percentage change (APC) and proportional mortality rates (PMR).ResultsFrom 2003-2017, breast cancer deaths comprised 14.4% in NHWs, 13.7% in aggregate AAs, 19.8% in Asian Indians, and 18.6% of all cancer deaths in Filipinas. While NHW breast cancer mortality rate significantly decreased (APC -2.1; CI -2.6, -1.6; p < 0.001) from 2003 to 2017, aggregate AA mortality rates were unchanged (APC 3.07; CI -0.37, 7.8; p = 0.071). However, when disaggregated, breast cancer mortality in Filipina (APC 1.9; CI 0.8, 3.0; p < 0.002), Chinese (APC 2.1; CI 1.3, 3.0; p < 0.001), and Korean (APC 2.6; CI 1.0, 4.1; p = 0.004) women significantly increased. Breast cancer mortality rates in Japanese women decreased (APC -1.9; CI -5.9, 2.1; p = 0.3).ConclusionWhile the proportion of women dying from breast cancer were similar in NHWs and aggregate Asians, when disaggregated, Filipina, Korean, and Chinese women had increased mortality rates over the past 15 years. During this time, breast cancer mortality in NHW and Japanese women decreased. Understanding disaggregated breast cancer mortality rates in Asians may improve culturally-tailored outreach, prevention, and treatment strategies to reduce cancer deaths from this critical disease.