Investigating facial shape using multilevel principal component analysis

SUMMARY Aims: 1. To determine the influence of geographical location, sex, height, Body Mass Index (BMI), age (14-16 years old), pubertal stage, metabolic factors, atopy, breathing disorders, maternal smoking and alcohol consumption during pregnancy on facial shape. 2. To explore the usefulness of Multilevel Principal Component Analysis (mPCA) in facial shape research. Method: The influence of geographical location and sex was assessed using 21 landmarks on 3D facial scans of subjects from Croatia (n=73), England (n=79), Wales (n=50) and Finland (n=47). The influence of sex, height, BMI, age (14-16 years old), pubertal stage, metabolic factors, atopy, breathing disorders, maternal smoking and alcohol consumption during pregnancy on adolescent facial shape was assessed using 1000 and 7160 quasi-landmarks on 3D facial scans of the Avon Longitudinal Study of Parents and Children (ALSPAC) cohort (n=1411). The results of mPCA were compared to those using landmarks only, conventional Principal Component Analysis (PCA), Discriminate Function Analysis (DFA) and Partial Least Squares Regression (PLSR). mPCA was also assessed as a variable selection tool prior to PLSR. Results: mPCA provided more meaningful information in the exploratory phase of data analysis than conventional PCA and DFA. However, the results must be interpreted with caution when group sizes are imbalanced. All variables reached significance, except for age, in their respective mPCA models. Geographical location, sex, height, BMI and fasting insulin explained greater than 5% of the total variation. These variables also reached significance in the PLSR models. Therefore 5% may be a useful threshold for PLSR variable selection. Conclusions: Sex, geographical location, height, BMI and fasting insulin had the most influence on facial shape. mPCA appears to be a useful tool for visualising the maximum variation between groups of subjects when group sizes are balanced and as a variable selection tool to inform more sophisticated models such as PLSR

Quantification of Progesterone and 17-β Estradiol in Mouse Serum by Liquid Chromatography-Tandem Mass Spectrometry

Quantification of progesterone and 17-β estradiol in mouse serum by liquid chromatography-tandem mass spectrometry Authors: Benjamin Kennard, Allison Cobble, Amy Gravitte, Keleigh Galloway, Jen Kintner, Jennifer Hall, Stacy Brown Introduction: In the United States, Chlamydia trachomatis is a commonly appearing sexually transmitted infection1. It affects the U.S. healthcare system to a tune of about $500 million dollars annually2. In women, it generally appears asymptomatic and can lead to severe secondary complications such as pelvic inflammatory diseases or infertility1. Female sex hormones, estrogen and progesterone, are being identified to have a role in chlamydial infection. Specifically, this study aims to create quantification methods to detect levels of estrogen and progesterone in mice, infected with Chlamydia muridarum, plasma samples. Methods: Progesterone samples were prepared using solid-liquid extraction (SLE+) cartridges with ethyl acetate as the elution solvent. Estradiol samples were prepared using liquid-liquid extraction (LLE) with methyl tert-butyl ether and subsequent derivatization with DMIS. Following sample preparation, hormones were quantified in samples using LC-MS/MS with a gradient elution of 1 mM ammonium fluoride in water and acetonitrile. The separation was achieved using a UCT C18 column (100 x 21.mm, 1.8 μm particle size) maintained at 50oC. The mass spectrometer was set up to isolate molecular ions for progesterone (m/z 315.0910) and derivatized estradiol (m/z 431.1835). Quantification was facilitated by the use of deuterium-labeled internal standards and their corresponding molecular ions in the mass spectrometer (d9-progesterone; m/z 324.1230 and d5-estradiol; m/z 436.2922). Results: Several aspects of the assay presented have been optimized for maximum analyte recovery and analytical sensitivity, including column choice, mobile phase, derivatizing agents for estradiol, and extraction protocols for progesterone. The LC-MS/MS method was investigated for precision and accuracy over three separate days. The dynamic range of the progesterone assay was 5 – 100 ng/mL, with a limit of detection of 1 ng/mL. Likewise, the estradiol assay was linear in the range of 5 – 100 ng/mL, with a limit of detection of 0.5 ng/mL. The average precision, represented by % RSD was 0.74 – 8.5% and 6.3 – 13.4% for progesterone and estradiol, respectively. The accuracy of the method, represented by % error was 1.6 – 14.4% and 4.0 – 10.5% for progesterone and estradiol, respectively. Successful validation was defined as \u3c 15% RSD and error (\u3c 20% at the limit of quantification), per current FDA Guidelines. Conclusions: The developed LC-MS/MS method is specific for progesterone and estradiol, and the extraction is suitable for preparation of mouse serum samples. This assay could be successfully applied to hormone quantification in mouse samples to support the investigation of the link between chlamydia infection and hormone levels in female animals. References 1. Chlamydia - 2017 Sexually Transmitted Diseases Surveillance. https://www.cdc.gov/std/stats17/chlamydia.htm. Accessed October 23, 2018. 2. Owusu-Edusei K, Chesson HW, Gift TL, et al. The Estimated Direct Medical Cost of Selected Sexually Transmitted Infections in the United States, 2008. Sex Transm Dis. 2013;40(3):197-201. doi:10.1097/OLQ.0b013e318285c6d

Mental health mobile app use: Considerations for serving underserved patients in integrated primary care settings

Depression, the most commonly experienced mental illness, affects more than 264 million people and is one of the leading causes of disability worldwide [1]. Depression, or major depressive disorder, is characterized by depressed mood and often accompanies other symptoms such as lack of interest, fatigue, feelings of worthlessness, impaired thinking, psychomotor agitation or slowing, thoughts of being better off dead and/or of suicide [2]. According to the 2018 National Survey of Drug Use and Health (NSDUH), 17.7 million or 7.7% of U.S. adults met the criteria for having a major depressive episode in the past year [3]. The prevalence of depression is closely related to socioeconomic factors and prevalence increases as family income level decreases. For example, 19.8% of women aged 20 years and older living below the federal poverty level (FPL) experience depression compared to only 4.8% of those living at or above 400% of the FPL [4]. Despite the availability of safe and effective treatments, many adults diagnosed with depressive disorders do not receive treatment [5]. Of the 17.7 million adults (≥ 18 years) meeting criteria for a major depressive disorder in the past year, only 64.8% reported having received treatment for depression [3]. There are disparities in receipt of treatment for mental health care, particularly among racial and ethnic minority groups. Only about 30% of African American and Hispanic adults (≤18 years) with any mental illness receive treatment, compared to 43.3% of the overall U.S. population [3]. Further, for those with lower education, the odds of receiving treatment declined (women) or remained stable (men) between 2005 and 2014 [6]

Binding of Elementary Bodies by the Opportunistic Fungal Pathogen Candida albicansor Soluble β-Glucan, Laminarin, Inhibits Chlamydia Trachomatisinfectivity

Microbial interactions represent an understudied facet of human health and disease. In this study, the interactions that occur between Chlamydia trachomatis and the opportunistic fungal pathogen, Candida albicans were investigated. Candida albicans is a common component of the oral and vaginal microbiota responsible for thrush and vaginal yeast infections. Normally, Candida exist in the body as yeast. However, disruptions to the microbiota create conditions that allow expanded growth of Candida, conversion to the hyphal form, and tissue invasion. Previous studies have shown that a myriad of outcomes can occur when Candida albicans interacts with pathogenic bacteria. To determine if C. trachomatis physically interacts with C. albicans, we incubated chlamydial elementary bodies (EB) in medium alone or with C. albicans yeast or hyphal forms for 1 h. Following incubation, the samples were formaldehyde-fixed and processed for immunofluorescence assays using anti-chlamydial MOMP or anti- chlamydial LPS antibodies. Replicate samples were replenished with culture medium and incubated at 35°C for 0-120 h prior to fixation for immunofluorescence analysis or collection for EB infectivity assays. Data from this study indicates that both C. trachomatis serovar E and C. muridarum EB bind to C. albicans yeast and hyphal forms. This interaction was not blocked by pre-incubation of EB with the Candida cell wall components, mannan or β-glucans, suggesting that EB interact with a Candida cell wall protein or other structure. Bound EB remained attached to C. albicans for a minimum of 5 days (120 h). Infectivity assays demonstrated that EB bound to C. albicans are infectious immediately following binding (0h). However, once bound to C. albicans, EB infectivity decreased at a faster rate than EB in medium alone. At 6h post binding, 40% of EB incubated in medium alone remained infectious compared to only 16% of EB bound to C. albicans. Likewise, pre-incubation of EB with laminarin, a soluble preparation of β-glucan, alone or in combination with other fungal cell wall components significantly decreases chlamydial infectivity in HeLa cells. These data indicate that interactions between EB and C. albicans inhibit chlamydial infectivity, possibly by physically blocking EB interactions with host cell receptors

A qualitative exploration of the feasibility of incorporating depression apps into integrated primary care clinics

Background: The use of mobile applications or “apps” is beginning to be identified as a potential cost-effective tool for treating depression. While the use of mobile apps for health management appears promising, little is known on how to incorporate these tools into integrated primary care settings – especially from the viewpoints of patients and the clinic personnel. Purpose: The purpose of this study was to explore patient- and clinic-level perceptions of the use of depression self-management apps within an integrated primary care setting. Methods: Patients (n=17), healthcare providers, and staff (n=15) completed focus groups or semi-structured interviews in-person or via Zoom between January and July 2020. Participants were asked about barriers and facilitators to app use, how to best integrate it into care, and reviewed pre-selected mental health apps. Data were analyzed using a directed content analysis approach. Results: From a patient perspective, features within the app such as notifications, the provision of information, easy navigation, and a chat/support function as well as an ability to share data with their doctor were desirable. Providers and staff identified integration of app data into electronic health records to be able to share data with patients and the healthcare team as well as clear evidence of effectiveness as factors that could facilitate implementation. All participants who reviewed apps identified at least one of them they would be interested in continuing to use. Conclusions: Overall, patients, healthcare providers, and staff believed depression apps could be beneficial for both patients and the clinic

Multilevel analysis of the influence of maternal smoking and alcohol consumption on the facial shape of English adolescents

This cross-sectional study aims to assess the influence of maternal smoking and alcohol consumption during pregnancy on the facial shape of non-syndromic English adolescents and demonstrate the potential benefits of using multilevel principal component analysis (mPCA). A cohort of 3755 non-syndromic 15-year-olds from the Avon Longitudinal Study of Parents and Children (ALSPAC), England, were included. Maternal smoking and alcohol consumption during the 1st and 2nd trimesters of pregnancy were determined via questionnaire at 18 weeks gestation. 21 facial landmarks, used as a proxy for the main facial features, were manually plotted onto 3D facial scans of the participants. The effect of maternal smoking and maternal alcohol consumption (average 1–2 glasses per week) was minimal, with 0.66% and 0.48% of the variation in the 21 landmarks of non-syndromic offspring explained, respectively. This study provides a further example of mPCA being used effectively as a descriptive analysis in facial shape research. This is the first example of mPCA being extended to four levels to assess the influence of environmental factors. Further work on the influence of high/low levels of smoking and alcohol and providing inferential evidence is required