Life course determinants of women’s health : from reproductive age to menopause

Over the past four decades, growing evidence has indicated that characteristics such as birth weight and length of gestation are not only key indicators for infant’s health, but also predictors of adult health and disease risk. These findings lend support to the developmental origins of health and disease theory. However, evidence remains inconclusive in terms of female hormone-related disorders, including endometriosis and perimenopausal disorders. Also, little is known about the social patterning of these female health burdens from population-based studies. Furthermore, the psychological health of women with endometriosis has not been adequately explored in longitudinal studies. Based on identified knowledge gaps and taking advantage of Swedish high-quality population-based registers, this thesis aims to study how factors operating during early life, such as parental and birth characteristics, adult socioeconomic and reproductive factors are associated with subsequent risks of endometriosis and perimenopausal disorders. Further, it explores whether women with endometriosis have higher risk of psychiatric comorbidity. Study I explored the associations of early life social and health characteristics with risk of endometriosis in a cohort of the second generation of women from the Uppsala Birth Cohort Multigenerational Study. Lower birth weight-for-gestational age, fewer births, and previous infertility disorder were found to be associated with an increased risk of endometriosis. Nevertheless, the inverse association between low birth weight-for-gestational age and endometriosis could not be explained by women’s lower number of live births in adulthood. Study II replicated the original findings in Study I in a nationwide population-based cohort of females born in Sweden between 1973 and 1987. This study confirmed the inverse association between fetal growth rate and risk of endometriosis, and expanded Study I by showing associations of maternal smoking during pregnancy and lower maternal education with endometriosis risk in early to mid-adulthood. The study also found a part of the association between maternal smoking and risk of early-onset of endometriosis was due to slow fetal growth. Study III focused on the psychological health of women with endometriosis by assessing the bi-directional associations of endometriosis with all psychiatric disorders, as well as the role of familial confounding, in a nationwide cohort of all women born in Sweden in 1973-1990. Statistically significant bi-directional associations were found for endometriosis with many different types of psychiatric disorders, including affective psychotic disorders, depressive, anxiety and stress-related disorders, eating disorders, alcohol/drug dependence, personality disorders, and attention-deficit hyperactivity disorder. These bi-directional associations observed at the population level largely remained in comparisons between exposed and unexposed sisters, suggesting that shared familial liability may not fully explain these associations. Study IV investigated the developmental origins of three subtypes of perimenopausal disorders using the same cohort as Study I. Positive association between birth weight and a clinical diagnosis of menopausal and climacteric states was found. Higher risk of being diagnosed with other perimenopausal disorders (e.g., atrophic vaginitis) was observed among women born with shorter gestational age. This study also documented that women with higher parental and own educational level in adulthood were more likely to be diagnosed with perimenopausal disorders. Taken together, this thesis supports the developmental origins of two important female hormone-related disorders during reproductive age and menopause, namely endometriosis and perimenopausal disorders. Our findings highlight the importance of intrauterine environment in shaping the developmental adaptations of metabolism and organ function. In addition to the developmental origins, these female health burdens were associated with a range of socioeconomic and reproductive factors as well as mental health in earlier life. It is therefore important to take a life-course perspective for a greater understanding of the etiology of hormone-related health outcomes and consider potential targeting of the high-risk groups for earlier public health intervention

The 2-Aminoethoxydiphenyl Borate Analog Dpb161 Blocks Storeoperated Ca 2+ Entry In Acutely Dissociated Rat Submandibular Cells

Cellular Ca 2+ signals play a critical role in cell physiology and pathology. In most non-excitable cells, store-operated Ca 2+ entry (SOCE) is an important mechanism by which intracellular Ca 2+ signaling is regulated. However, few drugs can selectively modulate SOCE. 2-Aminoethoxydiphenyl borate (2APB) and its analogs (DPB162 and DPB163) have been reported to inhibit SOCE. Here, we examined the effects of another 2-APB analog, DPB161 on SOCE in acutely-isolated rat submandibular cells. Both patch-clamp recordings and Ca 2+ imaging showed that upon removal of extracellular Ca 2+ ([Ca 2+ ] o =0), rat submandibular cells were unable to maintain ACh-induced Ca 2+ oscillations, but restoration of [Ca 2+ ] o to refill Ca 2+ stores enable recovery of these Ca 2+ oscillations. However, addition of 50 μM DPB161 with [Ca 2+ ] o to extracellular solution prevented the refilling of Ca 2+ store. Fura-2 Ca 2+ imaging showed that DPB161 inhibited SOCE in a concentration-dependent manner. After depleting Ca 2+ stores by thapsigargin treatment, bath perfusion of 1 mM Ca 2+ induced [Ca 2+ ] i elevation in a manner that was prevented by DPB161. Collectively, these results show that the 2-APB analog DPB161 blocks SOCE in rat submandibular cells, suggesting that this compound can be developed as a pharmacological tool for the study of SOCE function and as a new therapeutic agent for treating SOCE-associated disorders

Proinflammatory allogeneic dendritic cells enhance the therapeutic efficacy of systemic anti-4-1BB treatment

As an immune adjuvant, proinflammatory allogeneic dendritic cells (AlloDCs) have demonstrated promising immune-priming effects in several preclinical and clinical studies. The effector cells, including NK cells and T cells are widely acknowledged as pivotal factors in the effectiveness of cancer immunotherapy due to their ability to selectively identify and eradicate malignant cells. 4-1BB, as a costimulatory receptor, plays a significant role in the stimulation of effector cell activation. This study evaluated the anti-tumor effects when combining intratumoral administration of the immune-adjuvant AlloDCs with systemic α4-1BB treatment directly acting on effector cells. In both the CT-26 murine colon carcinoma model and B16 murine melanoma model, AlloDCs demonstrated a significant enhancement in the therapeutic efficacy of α4-1BB antibody. This enhancement was observed through the delayed growth of tumors and prolonged survival. Analysis of the tumor microenvironment (TME) in the combined-treatment group revealed an immune-inflamed TME characterized by increased infiltration of activated endogenous DCs and IFNγ+ CD8+ T cells, showing reduced signs of exhaustion. Furthermore, there was an augmented presence of tissue-resident memory (TRM) CD8+ T cells (CD103+CD49a+CD69+). The combination treatment also led to increased infiltration of CD39+CD103+ tumor-specific CD8+ T cells and neoantigen-specific T cells into the tumor. Additionally, the combined treatment resulted in a less immunosuppressive TME, indicated by decreased infiltration of myeloid-derived suppressor cells and Tregs. These findings suggest that the combination of intratumoral AlloDCs administration with systemic agonistic α4-1BB treatment can generate a synergistic anti-tumor response, thereby warranting further investigation through clinical studies

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data