Multiple reaction monitoring assays for large-scale quantitation of proteins from 20 mouse organs and tissues.

Mouse is the mammalian model of choice to study human health and disease due to its size, ease of breeding and the natural occurrence of conditions mimicking human pathology. Here we design and validate multiple reaction monitoring mass spectrometry (MRM-MS) assays for quantitation of 2118 unique proteins in 20 murine tissues and organs. We provide open access to technical aspects of these assays to enable their implementation in other laboratories, and demonstrate their suitability for proteomic profiling in mice by measuring normal protein abundances in tissues from three mouse strains: C57BL/6NCrl, NOD/SCID, and BALB/cAnNCrl. Sex- and strain-specific differences in protein abundances are identified and described, and the measured values are freely accessible via our MouseQuaPro database: http://mousequapro.proteincentre.com . Together, this large library of quantitative MRM-MS assays established in mice and the measured baseline protein abundances represent an important resource for research involving mouse models.

Process and Workflow for Preparation of Disparate Mouse Tissues for Proteomic Analysis.

We investigated the effect of homogenization strategy and protein precipitation on downstream protein quantitation using multiple reaction monitoring mass spectrometry (MRM-MS). Our objective was to develop a workflow capable of processing disparate tissue types with high throughput, minimal variability, and maximum purity. Similar abundances of endogenous proteins were measured in nine different mouse tissues regardless of the homogenization method used; however, protein precipitation had strong positive effects on several targets. The best throughput was achieved by lyophilizing tissues to dryness, followed by homogenization via bead-beating without sample buffer. Finally, the effect of tissue perfusion prior to dissection and collection was explored in 20 mouse tissues. MRM-MS showed decreased abundances of blood-related proteins in perfused tissues; however, complete removal was not achieved. Concentrations of nonblood proteins were largely unchanged, although significantly higher variances were observed for proteins from the perfused lung, indicating that perfusion may not be suitable for this organ. We present a simple yet effective tissue processing workflow consisting of harvest of fresh nonperfused tissue, novel lyophilization and homogenization by bead-beating, and protein precipitation. This workflow can be applied to a range of mouse tissues with the advantages of simplicity, minimal manual manipulation of samples, use of commonly available equipment, and high sample quality.

Antibacterial Copper-Doped Calcium Phosphate Glasses for Bone Tissue Regeneration.

Calcium phosphate glasses are a promising new generation of biomaterials that can simultaneously induce tissue regeneration and controlled release of therapeutic molecules. In this work, novel calcium phosphate glasses containing 0, 2, 4, and 6 mol % Cu2+ were synthesized via room temperature precipitation reaction in aqueous solution. The effect of Cu2+ addition on the glass properties and structure was investigated using thermal analysis, 31P solid-state MAS NMR, Raman spectroscopy, and X-ray diffraction. All glasses crystallize at temperature >500 °C and are mainly formed by Q1 groups. The release of P, Ca, and Cu in solution over time was monitored via inductively coupled plasma-optical emission spectroscopy. It was found that with increasing Cu content, the amount of P and Ca released decreases whereas the amount of Cu released increases. The effect of Cu2+ release on the antibacterial activity against S. aureus, a bacterial strain commonly found in postsurgery infections, has been investigated. The addition of copper has been shown to infer the glasses antibacterial properties. As expected, the antibacterial activity of the glasses increases with increasing Cu2+ content. Cytocompatibility was assessed by seeding human osteoblast-like osteosarcoma cells Saos-2 (HTB85) on the glass particles. A significant increase in cell number was observed in all the glasses investigated. The copper-doped calcium phosphate glasses have proven to be multifunctional, as they combine bone regenerative properties with antibacterial activity. Therefore, they have great potential as antibacterial bioresorbable materials for hard tissue regeneration.

Disparities in Counseling Female Cancer Patients for Fertility Preservation.

BACKGROUND: Female cancer patients who are exposed to gonadotoxic chemotherapy are at risk of future infertility. Research suggests that disparities in fertility preservation counseling (FPC) may exist. Previous research is limited by recall bias; therefore, this study examined objective electronic medical chart data regarding FPC at an academic medical center. MATERIALS AND METHODS: This study included reproductive-aged women (18-45 years old) with a diagnosis of breast, gynecological, or hematological cancer and who were exposed to a gonadotoxic chemotherapeutic agent from 2009 to 2013. Chi-square and logistic regression analyses were utilized to analyze disparities in FPC. RESULTS: Two hundred fifty-nine women met the study criteria. One hundred eighty-one women were diagnosed with breast cancer, 52 with hematological cancer, and 26 with gynecological cancer. 160/259 (62%) women had documented counseling for fertility preservation (FP), 60 (23%) women were not counseled as counseling was determined to be "not applicable," 16 (6%) women were not counseled and no explanation was given for the lack of counseling, and counseling was not documented in 23 (9%) charts. Age, marital status, and racial/ethnic background were related to counseling status. Patients with gynecological or hematological cancer were more likely to be counseled than other patients. Logistic regression results demonstrated that FPC was largely driven by cancer diagnosis. CONCLUSIONS: Although cancer diagnosis was the greatest predictor of FPC, disparities were evident in the counseling of female cancer patients for FP treatment. Equality in counseling female patients for FP treatment is imperative to reduce the risk of emotional harm and future infertility.

Social and Environmental Barriers to Nutrition Therapy for Diabetes Management Among Underserved Pregnant Women: A Qualitative Analysis.

OBJECTIVE: To identify social and environmental barriers to nutrition therapy for diabetes management during pregnancy among a population of low-income, minority pregnant women. DESIGN: Prospective, in-depth, semi-structured interviews performed serially during pregnancy and continued until thematic saturation was reached. SETTING: Urban academic medical center. PARTICIPANTS: Ten pregnant low-income, minority women with gestational diabetes and type 2 diabetes mellitus. PHENOMENON OF INTEREST: Social and environmental barriers to nutrition therapy for diabetes management during pregnancy. ANALYSIS: Qualitative analysis of interview data using electronic coding software was performed using theme analysis. RESULTS: Participants perceived limited ability and self-efficacy to adopt nutrition recommendations. Specific themes identified as barriers included (1) feeling overwhelmed by the unfamiliar; (2) using and decoding nutrition labels; (3) managing nutrition choices and seeking control in the setting of food insecurity; (4) experiencing lack of control and motivation, and limited self-efficacy; (5) balancing recommendations with taste preferences and cultural food norms; (6) maintaining a healthy eating schedule; and (7) accommodating diabetes in family and social life. CONCLUSIONS AND IMPLICATIONS: Pregnant women with diabetes encounter a number of knowledge-based, attitudinal, and resource-related barriers that reduce capacity for nutrition therapy adherence. Provision of culturally informed, practical nutrition support that addresses the needs of women in low-resource communities is an important component of comprehensive diabetes care during pregnancy.

"I Was Tired of All the Sticking and Poking": Identifying Barriers to Diabetes Self-Care Among Low-Income Pregnant Women.

OBJECTIVE: Diabetes in pregnancy is a significant problem for low-income, minority women. We sought to evaluate barriers to diabetes self-care during pregnancy in an underserved population. METHODS: Twenty-nine in-depth, semi-structured interviews were performed over 10 women's pregnancies to identify barriers to successful diabetes management, using cognitive load theory to frame interview questions. Qualitative analysis of interview data used grounded theory techniques. RESULTS: Fifty percent of this cohort of minority, low-income, public aid-supported women had pregestational diabetes. Six barrier domains were identified: diabetes novelty; social and economic chaos; nutrition challenges; psychological stressors; burden of disease management; and outcome expectation inability to promote behavior change. CONCLUSIONS: Low-income women face multiple barriers to successful diabetes self-care during pregnancy. Ability to achieve diabetes goals in pregnancy is influenced by several social, cognitive, and knowledge-based factors. Understanding this complex interplay of factors impacting diabetes management may help providers work with patients in achieving healthy pregnancies.

Using community-based participatory research (CBPR) to develop a community-level HIV prevention intervention for Latinas: a local response to a global challenge.

INTRODUCTION AND BACKGROUND: The arsenal of interventions to reduce the disproportionate rates of HIV and sexually transmitted disease (STD) infection among Latinos in the United States lags behind what is available for other populations. The purpose of this project was to develop an intervention that builds on existing community strengths to promote sexual health among immigrant Latinas. METHODS: Our community-based participatory research (CBPR) partnership engaged in a multistep intervention development process. The steps were to (1) increase Latina participation in the existing partnership, (2) establish an intervention team, (3) review the existing sexual health literature, (4) explore health-related needs and priorities of Latinas, (5) narrow priorities based on what is important and changeable, (6) blend health behavior theory with Latinas' lived experiences, (7) design an intervention conceptual model, (8) develop training modules and (9) resource materials, and (10) pretest and (11) revise the intervention. RESULTS: The MuJEReS intervention contains five modules to train Latinas to serve as lay health advisors (LHAs) known as "Comadres." These modules synthesize locally collected data with other local and national data, blend health behavior theory with the lived experiences of immigrant Latinas, and harness a powerful existing community asset, namely, the informal social support Latinas provide one another. CONCLUSION: This promising intervention is designed to meet the sexual health priorities of Latinas. It extends beyond HIV and STDs and frames disease prevention within a sexual health promotion framework. It builds on the strong, preexisting social networks of Latinas and the preexisting, culturally congruent roles of LHAs.

Latino men who have sex with men and HIV in the rural south-eastern USA: findings from ethnographic in-depth interviews.

A community-based participatory research partnership explored HIV risk and potentially effective intervention characteristics to reduce exposure and transmission among immigrant Latino men who have sex with men living in the rural south-eastern USA. Twenty-one participants enrolled and completed a total of 62 ethnographic in-depth interviews. Mean age was 31 (range 18-48) years and English-language proficiency was limited; 18 participants were from Mexico. Four participants reported having sex with men and women during the past three months; two participants self-identified as male-to-female transgender. Qualitative themes that emerged included a lack of accurate information about HIV and prevention; the influence of social-political contexts to sexual risk; and barriers to healthcare services. We also identified eight characteristics of potentially effective interventions for HIV prevention. Our findings suggest that socio-political contexts must be additional targets of change to reduce and eliminate HIV health disparities experienced by immigrant Latino men who have sex with men.