RESUMO
STUDY QUESTION: How do plasma progesterone (P) and dydrogesterone (D) concentrations together with endometrial histology, transcriptomic signatures, and immune cell composition differ when oral dydrogesterone (O-DYD) or micronized vaginal progesterone (MVP) is used for luteal phase support (LPS)? SUMMARY ANSWER: Although after O-DYD intake, even at steady-state, plasma D and 20αdihydrodydrogesterone (DHD) concentrations spiked in comparison to P concentrations, a similar endometrial signature was observed by histological and transcriptomic analysis of the endometrium. WHAT IS KNOWN ALREADY: O-DYD for LPS has been proven to be noninferior compared to MVP in two phase III randomized controlled trials. Additionally, a combined individual participant data and aggregate data meta-analysis indicated that a higher pregnancy rate and live birth rate may be obtained in women receiving O-DYD versus MVP for LPS in fresh IVF/ICSI cycles. Little data are available on the pharmacokinetic (PK) profiles of O-DYD versus MVP and their potential molecular differences at the level of the reproductive organs, particularly at the endometrial level. STUDY DESIGN, SIZE, DURATION: Thirty oocyte donors were planned to undergo two ovarian stimulation (OS) cycles with dual triggering (1.000 IU hCG + 0.2 mg triptorelin), each followed by 1 week of LPS: O-DYD or MVP, in a randomized, cross-over, double-blind, double-dummy fashion. On both the first and eighth days of LPS, serial blood samples upon first dosing were harvested for plasma D, DHD, and P concentration analyses. On Day 8 of LPS, an endometrial biopsy was collected for histologic examination, transcriptomics, and immune cell analysis. PARTICIPANTS/MATERIALS, SETTING, METHODS: All oocyte donors were <35 years old, had regular menstrual cycles, no intrauterine contraceptive device, anti-Müllerian hormone within normal range and a BMI ≤29 kg/m2. OS was performed on a GnRH antagonist protocol followed by dual triggering (1.000 IU hCG + 0.2 mg triptorelin) as soon as ≥3 follicles of 20 mm were present. Following oocyte retrieval, subjects initiated LPS consisting of MVP 200 mg or O-DYD 10 mg, both three times daily. D, DHD, and P plasma levels were measured using liquid chromatography-tandem mass spectrometry. Histological assessment was carried out using the Noyes criteria. Endometrial RNA-sequencing was performed for individual biopsies and differential gene expression was analyzed. Endometrial single-cell suspensions were created followed by flow cytometry for immune cell typing. MAIN RESULTS AND THE ROLE OF CHANCE: A total of 21 women completed the entire study protocol. Subjects and stimulation characteristics were found to be similar between groups. Following the first dose of O-DYD, the average observed maximal plasma concentrations (Cmax) for D and DHD were 2.9 and 77 ng/ml, respectively. The Cmax for D and DHD was reached after 1.5 and 1.6 h (=Tmax), respectively. On the eighth day of LPS, the first administration of that day gave rise to a Cmax of 3.6 and 88 ng/ml for D and DHD, respectively. For both, the observed Tmax was 1.5 h. Following the first dose of MVP, the Cmax for P was 16 ng/ml with a Tmax of 4.2 h. On the eighth day of LPS, the first administration of that day showed a Cmax for P of 21 ng/ml with a Tmax of 7.3 h. All 42 biopsies showed endometrium in the secretory phase. The mean cycle day was 23.9 (±1.2) in the O-DYD group versus 24.0 (±1.3) in the MVP group. RNA-sequencing did not reveal significantly differentially expressed genes between samples of both study groups. The average Euclidean distance between samples following O-DYD was significantly lower than following MVP (respectively 12.1 versus 18.8, Mann-Whitney P = 6.98e-14). Immune cell profiling showed a decrease of CD3 T-cell, γδ T-cell, and B-cell frequencies after MVP treatment compared to O-DYD, while the frequency of natural killer (NK) cells was significantly increased. LIMITATIONS, REASONS FOR CAUTION: The main reason for caution is the small sample size, given the basic research nature of the project. The plasma concentrations are best estimates as this was not a formal PK study. Whole tissue bulk RNA-sequencing has been performed not correcting for bias caused by different tissue compositions across biopsies. WIDER IMPLICATIONS OF THE FINDINGS: This is the first study comparing O-DYD/MVP, head-to-head, in a randomized design on a molecular level in IVF/ICSI. Plasma serum concentrations suggest that administration frequency is important, in addition to dose, specifically for O-DYD showing a rapid clearance. The molecular endometrial data are overall comparable and thus support the previously reported noninferior reproductive outcomes for O-DYD as compared to MVP. Further research is needed to explore the smaller intersample distance following O-DYD and the subtle changes detected in endometrial immune cells. STUDY FUNDING/COMPETING INTEREST(S): Not related to this work, C.Bl. has received honoraria for lectures, presentations, manuscript writing, educational events, or scientific advice from Abbott, Ferring, Organon, Cooper Surgical, Gedeon-Richter, IBSA, and Merck. H.T. has received honoraria for lectures, presentations, manuscript writing, educational events, or scientific advice from Abbott, Ferring, Cooper Surgical, Gedeon-Richter, Cook, and Goodlife. S.M. has received honoraria for lectures, presentations, educational events, or scientific advice from Abbott, Cooper Surgical, Gedeon-Richter, IBSA, and Merck and Oxolife. G.G. has received honoraria for lectures, presentations, educational events, or scientific advice from Merck, MSD, Organon, Ferring, Theramex, Gedeon-Richter, Abbott, Biosilu, ReprodWissen, Obseva, PregLem, Guerbet, Cooper, Igyxos, and OxoLife. S.V.-S. is listed as inventor on two patents (WO2019115755A1 and WO2022073973A1), which are not related to this work. TRIAL REGISTRATION NUMBER: EUDRACT 2018-000105-23.
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Didrogesterona , Progesterona , Gravidez , Humanos , Feminino , Adulto , Estudos Cross-Over , Pamoato de Triptorrelina , Fase Luteal , Lipopolissacarídeos , Injeções de Esperma Intracitoplásmicas/métodos , Taxa de Gravidez , Indução da Ovulação/métodos , Endométrio , RNA , Fertilização in vitro/métodos , Ensaios Clínicos Controlados Aleatórios como AssuntoRESUMO
BACKGROUND: Pregnancy-related anxiety (PrA) has been identified as a construct distinct from general stress and anxiety with a negative impact on birth and child outcomes. Validated instruments with good psychometric properties to assess pregnancy-related anxiety in German-speaking expectant mothers are still lacking. The Pregnancy-Related Anxiety Questionnaire revised for its use independent of parity (PRAQ-R2) assesses fear of giving birth (FoGB), worries of bearing a physically or mentally handicapped child (WaHC) and concerns about own appearance (CoA). The aim of this study was to investigate the psychometric properties of the PRAQ-R2 in a German sample of pregnant women in their third pregnancy trimester. METHODS: The PRAQ-R2 and several questionnaires measuring different forms of anxiety as well as depressive symptoms and perceived general self-efficacy were administered cross-sectionally in a sample of nulliparous and parous women (N = 360) in the third trimester of pregnancy. RESULTS: Reliability was satisfactory to excellent for the PRAQ-R2 total scale (Cronbach's α = .85) and the subscales (α = .77 to .90). Confirmatory and exploratory factor analysis confirmed the three-factorial structure of the instrument. The three factors together explained 68% of variance. Construct validity was confirmed by positive low- to moderate-sized correlations of the PRAQ-R2 total score and the subscales with measurements of anxiety and depression and by negative low correlations with general self-efficacy. CONCLUSIONS: The German version of the PRAQ-R2 is a valid and feasible measurement for pregnancy-related anxiety for research and clinical practice.
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Ansiedade/diagnóstico , Depressão/diagnóstico , Complicações na Gravidez/diagnóstico , Terceiro Trimestre da Gravidez/psicologia , Inquéritos e Questionários/normas , Adulto , Medo/psicologia , Feminino , Alemanha , Humanos , Parto/psicologia , Gravidez , Psicometria , Reprodutibilidade dos Testes , Autoeficácia , Traduções , Adulto JovemRESUMO
OBJECTIVES: To investigate the effect of obesity in early-mid pregnancy on crucial pregnancy hormones and the uterine immune environment. BACKGROUND: Obesity impacts reproductive ability, adversely affecting conception and leading to complications in pregnancy. Obesity is often regarded as a stress state and an immune disease, both of which may contribute to pregnancy failure. We previously demonstrated that stress in early pregnancy greatly alters progesterone secretion. As progesterone is an immunomodulator, altered progesterone secretion may adversely modify the maternal immune system. In the current study, we test the hypothesis that obesity during pregnancy adversely alters the uterine immune environment. METHODS: An obese mouse model was created by feeding C57/BL6 mice on a high-fat (HF)/sugar diet for 12 weeks before pregnancy. Control mice were fed on lower-fat/sugar chow. Mice were mated, and on day 7.5 of pregnancy plasma progesterone and prolactin were measured by immunoassay. Cells from the uterus-draining inguinal lymph nodes were collected for analysis of the uterine immune response by flow cytometry. RESULTS: Diet-induced obesity increased the secretion of progesterone and altered a number of uterine natural killer (NK)- and T-cell responses. These included a marked reduction in the percentage of leucocyte-derived NK cells and reduced expression of interferon-γ (IFN-γ) in the NK cells compared with control mice. CONCLUSIONS: Maternal obesity, induced by an HF diet, may lead to a reduction in the expression of IFN-γ in NK cells. NK-cell-derived IFN-γ is reported to be involved in supporting uterine spiral artery remodelling. Thus, obesity in early pregnancy may compromise vascularization by reducing the expression of IFN-γ-positive NK cells. Furthermore, the expression of uterine CD8(+) cells was reduced in the HF diet-fed mice, suggesting obesity may adversely alter the maternal immune adaptation that is essential for effective pregnancy.
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Dieta Hiperlipídica/efeitos adversos , Interferon gama/metabolismo , Células Matadoras Naturais/metabolismo , Obesidade/patologia , Progesterona/metabolismo , Útero/patologia , Análise de Variância , Animais , Feminino , Feto/imunologia , Regulação da Expressão Gênica no Desenvolvimento , Tolerância Imunológica , Troca Materno-Fetal , Camundongos , Camundongos Endogâmicos C57BL , Obesidade/complicações , Obesidade/imunologia , Gravidez , Útero/imunologiaRESUMO
An increasing incidence of chronic immune diseases such as allergies, multiple sclerosis, and type 2 diabetes, as well as obesity and cardiovascular and psychiatric disorders has been reported over the last five decades. Since the human genome has not altered significantly over this period of time, gene-environment interactions are suspected to be responsible for these increased disease incidences. In this context, the prenatal period is believed to significantly contribute to altered disease susceptibilities, which could be associated with environmental factors to which pregnant women were exposed to. This observation has led to a concept entitled 'developmental origin of health and disease', a topic that is enjoying much attention in clinical and basic science research. The aim of these research endeavors is to postulate guidelines for primary disease prevention. Whilst the emerging insights from this field of research provide significant pieces of the puzzle, one area is still largely neglected: the clear identification of a sex-specific programming effect. Thus it is essential that such an approach becomes fully integrated in future research goals.
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Doença Crônica , Desenvolvimento Embrionário/genética , Epigênese Genética/genética , Interação Gene-Ambiente , Predisposição Genética para Doença/genética , Modelos Genéticos , Feminino , Humanos , Masculino , Caracteres Sexuais , Fatores SexuaisRESUMO
Subclinical depressive symptoms constitute a primary risk factor for major depression as well as for cardiovascular conditions, which may be mediated by endocrine or immune alterations. The aim of this study was to assess the association between the extent of subclinical depressive symptoms and neuroendocrine and immune cell responses to acute psychosocial stress in healthy females. In N = 33 healthy premenopausal women, state anxiety, plasma adrenocorticotropic hormone and serum cortisol, and interleukin-6 (IL-6) concentration responses to public speaking stress were assessed. Beck depression inventory (BDI) scores were entered as a covariate in the analyses. The IL-6 response was significantly associated with BDI scores (p < 0.05). Secondary analyses revealed that women with more subclinical depressive symptoms demonstrated a reduced stress-induced increase in circulating IL-6 level (p < 0.05). By contrast, stress-induced neuroendocrine activation was not associated with depressive symptoms. Hence, subclinical depressive symptoms were associated with IL-6 responses to stress in young, healthy women. Unexpectedly, there was a reduced increase of serum IL-6 level in response to stress. Effects of depressive symptoms on the IL-6 response to stress may differ between subclinical and major depression.
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Depressão/psicologia , Estresse Psicológico/sangue , Hormônio Adrenocorticotrópico/sangue , Adulto , Ansiedade , Depressão/sangue , Depressão/imunologia , Transtorno Depressivo Maior , Feminino , Humanos , Hidrocortisona/sangue , Interleucina-6/sangue , Pré-Menopausa , Estresse Psicológico/imunologiaRESUMO
Successful blastocyst implantation requires intricately orchestrated adaptation processes involving maternal and fetal mediators. The pivotal role of distinct immune response pathways in early pregnancy is widely acknowledged. Pro-inflammatory cytokines, e.g. interferon-gamma (IFN-gamma), are the primary inducers of tryptophan-degrading enzyme indoleamine 2,3-dioxygenase (IDO) and of neopterin biosynthesis by GTP-cyclohydrolase I. IDO activity has been proposed to be of high clinical relevance in the context of pregnancy. To date, insights arising from clinical studies on IDO activity and neopterin concentration during the very early days of pregnancy are still few. Early morning urinary neopterin concentrations in 61 women undergoing assisted reproduction treatment (72 cycles in total) were examined, upon exclusion of infections, daily over a period of 2 weeks after embryo transfer. Twenty of the study participants (28%) became successfully pregnant, and four women experienced abortion. Neopterin concentrations significantly increased after blastocyst transfer when implantation was successful (chi-squared=23.291, P<0.01; Friedman test), opposed to non-significant changes of neopterin in women with unsuccessful treatment (chi-squared=8.203). The steady increase of neopterin concentrations upon blastocyst transfer indicates that heightened production of neopterin in very early phases of pregnancy may serve as an early predictor of successfully progressing pregnancies in humans.
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Biomarcadores/urina , Implantação do Embrião , Neopterina/urina , Técnicas de Reprodução Assistida , Adulto , Feminino , Humanos , GravidezRESUMO
OBJECTIVE: To analyze the neuroendocrine and immune cell responses to acute psychosocial stress in obese compared to non-obese premenopausal women. METHODS: N=15 obese (BMI> or =30) and N=24 (BMI<30) non-obese premenopausal women underwent public speaking stress. State anxiety, ACTH, cortisol, and the redistribution of immune cells were measured before, during, and 10 and 45min after public speaking. Serum hsCRP and serum IL-6 levels were analyzed before, and IL-6 additionally 45min after stress. RESULTS: In response to public speaking stress, both groups showed significant but comparable increases in state anxiety, plasma ACTH, and blood pressure (all p<0.01; time effects). The cortisol stress response was significantly enhanced in obese women (p<0.05; interaction effect). In addition, heart rate and diastolic blood pressure were significantly higher in obese women 10min following stress (p<0.05, t-tests). Public speaking stress led to a significant increase in IL-6 concentrations (p<0.001; time effect), and obese women displayed higher IL-6 levels both pre- and post-stress (p<0.05; group effect; between-group t-tests: pre-stress p<0.05; post-stress p<0.01). Baseline numbers of circulating leukocytes, granulocytes, CD3+ cells and hsCRP concentration were significantly higher in obese women (between-group t-tests: all p<0.05, but the groups did not differ in the stress-induced redistribution of circulating leukocyte subpopulations. CONCLUSIONS: Our data reveal a strong association of obesity with chronic low-grade inflammation in premenopausal women. This pro-inflammatory state, together with altered neuroendocrine and cardiovascular stress responsiveness, may conceivably constitute one of the mechanisms linking psychological stress and the long-term health risks associated with obesity.
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Hemodinâmica , Imunidade Celular , Obesidade/fisiopatologia , Estresse Psicológico/fisiopatologia , Hormônio Adrenocorticotrópico/sangue , Adulto , Pressão Sanguínea , Proteína C-Reativa/metabolismo , Estudos de Casos e Controles , Feminino , Frequência Cardíaca , Humanos , Hidrocortisona/sangue , Interleucina-6/sangue , Obesidade/complicações , Pré-Menopausa , Fala , Estresse Psicológico/complicaçõesRESUMO
BACKGROUND: Stress has been suggested to impact the onset and exacerbation of eczema and other atopic disorders. Whether early exposure to stress-related factors might exert long-term effects remains to be clarified. OBJECTIVE: The objective of this study was to investigate whether stress-related maternal factors during pregnancy are associated with childhood eczema during the first 6 years of life. METHODS: Data from 3004 children from a prospective German birth cohort study (LISA) were analyzed. Information from maternity certificates and questionnaire information on unwanted pregnancy were used to evaluate stress-related maternal factors during pregnancy. Prevalence data for physician-diagnosed eczema were available up to the age of 6 years. RESULTS: Maternal factors during pregnancy were positively associated with childhood eczema in terms of cumulative prevalence up to the age of 2 years (adjusted odds ratio, 1.48; 95% confidence interval, 0.95-2.30) after adjusting for potential confounders. Beyond the second year no increased risk was observed. CONCLUSIONS: The results of this study suggest that stress-related maternal factors during pregnancy are associated with an increased risk of childhood eczema during the first 2 years of life. The impact of postnatal stress such as parental divorce or separation on this association could not be clarified. Future studies should therefore further elucidate how prenatal and postnatal stress interact and whether prenatal stress might have a programming effect. If future studies confirm the findings of this study, reducing maternal stress during pregnancy might be a possible target in the primary prevention of eczema during childhood.
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Eczema/epidemiologia , Troca Materno-Fetal/fisiologia , Complicações na Gravidez , Estresse Fisiológico , Estresse Psicológico , Adulto , Criança , Pré-Escolar , Estudos de Coortes , Eczema/congênito , Feminino , Seguimentos , Alemanha , Humanos , Imunomodulação , Lactente , Recém-Nascido , Masculino , Gravidez , Prevalência , Fatores de RiscoRESUMO
BACKGROUND: In polycystic ovary syndrome (PCOS), one of the main features is chronic anovulation associated with lower pregnancy rates. Little is known regarding the psychological aspects associated with infertility in these patients. Therefore, we examined the influence of an unfulfilled wish to conceive on various aspects of psychological functioning in PCOS women. METHODS: Standardized questionnaires assessing quality-of-life (36-item short-form health survey, SF-36), depressiveness (Beck Depression Inventory), emotional distress (Symptom Check List 90, SCL-90-R), sexual satisfaction and self-worth (visual analogue scales), and a questionnaire on the desire for a child (FKW) were administered at the outpatient endocrine clinic to consecutive PCOS patients. RESULTS: Questionnaires from 115 PCOS patients were analysed. The majority (76.1%) worried about remaining childless in the future, and 51.3% reported a current wish to conceive. 23.9% of patients had scores indicating mild to moderate depression, and 25.2% had scores indicating clinically relevant depression. Furthermore, all quality-of-life scores were significantly lower compared with normative data (P < 0.001). Unexpectedly, comparisons of patients with a current unfulfilled desire to conceive to those with no present wish for a child revealed no discernable impact on depressive symptoms, quality-of-life or emotional distress. Reduced sexual satisfaction and self-worth were largely determined by partnership status and not infertility. However for PCOS patients who wished to conceive, the wish for a child was a significantly greater priority when compared with normative data from infertile patients. CONCLUSIONS: PCOS represents a major risk factor for psychosocial and emotional problems, but at least in this sample of PCOS patients, infertility does not appear to constitute a primary determinant of psychological problems.
Assuntos
Infertilidade Feminina/psicologia , Transtornos Mentais/complicações , Síndrome do Ovário Policístico/complicações , Adulto , Coito/psicologia , Depressão/complicações , Feminino , Humanos , Infertilidade Feminina/etiologia , Satisfação Pessoal , Qualidade de Vida , Fatores de Risco , Autoimagem , Estresse PsicológicoRESUMO
BACKGROUND AND OBJECTIVE: Women with polycystic ovary syndrome (PCOS) present with multiple risk factors for cardiovascular diseases at a young age, including obesity and chronic low-grade inflammation. Since depression is common in PCOS, this study aimed to address whether depression correlates with indices of chronic low-grade inflammation beyond the association with obesity. METHODS: Serum concentrations of IL-6, the stimulated production of IFN-gamma, TNF-alpha, IL-2, IL-4, IL-5, and IL-10, leukocyte numbers, and hsCRP were analyzed in 57 PCOS patients and 28 healthy women, together with clinical parameters, including body mass index (BMI), testosterone, and insulin resistance (HOMA-IR), and psychological parameters, including Beck Depression Inventory (BDI) and health-related quality-of-life (SF-36) scores. RESULTS: PCOS patients demonstrated significantly increased hsCRP, IL-6, and leukocyte numbers. Group differences in IL-6 and leukocyte numbers, but not hsCRP, disappeared after controlling for BMI. The stimulated production of IFN-gamma, TNF-alpha, IL-2, IL-4, and IL-5 was significantly decreased, irrespective of BMI. In PCOS, hsCRP, IL-6, and leukocyte numbers were correlated with BMI, HDL, diastolic blood pressure, and with insulin resistance. On the other hand, no correlations were found with depression scores or with PCOS-specific endocrine abnormalities. In regression models, BMI was a significant predictor of the key immune markers, and explained a large amount of variance, whereas BDI was not included in either model. CONCLUSIONS: These data confirm that obesity plays a pivotal role in inflammatory processes relevant to cardiovascular risk in women with PCOS. However, even lean PCOS patients may display subtle alterations in specific aspects of immunity. Our findings did not support a correlation of depression with chronic low-grade inflammation in PCOS.
Assuntos
Transtorno Depressivo/epidemiologia , Inflamação/epidemiologia , Obesidade/epidemiologia , Síndrome do Ovário Policístico/epidemiologia , Adulto , Biomarcadores/sangue , Índice de Massa Corporal , Doenças Cardiovasculares/epidemiologia , Doença Crônica , Estudos Transversais , Transtorno Depressivo/imunologia , Transtorno Depressivo/psicologia , Feminino , Humanos , Inflamação/imunologia , Inflamação/psicologia , Obesidade/imunologia , Obesidade/psicologia , Síndrome do Ovário Policístico/imunologia , Síndrome do Ovário Policístico/psicologia , Valor Preditivo dos Testes , Fatores de RiscoRESUMO
OBJECTIVE: Prenatal distress has been linked to pregnancy complications and poor offspring's health, despite the fact that longitudinal assessments of various stress dimensions are still lacking. Hence, we aimed to assess perceived stress over the course of pregnancy. Moreover, we examined whether social support and coping styles are linked to prenatal stress trajectories. METHODS: Data from 543 women participating in the PRINCE (Prenatal Identification of Children Health) study, a prospective population-based cohort study, was used for the present analyses. Once per trimester the women completed questionnaires regarding different psychometric measures, including the Perceived Stress Scale (PSS). Linear mixed regression models were used to examine perceived stress development longitudinally and to relate social support and coping styles to stress trajectories during pregnancy. RESULTS: A significant decrease of perceived stress was observed over the course of pregnancy. Stratifying the study sample according to parity, women delivering their first child had continuously lower perceived stress scores compared to women having already one or more children, and a significant decrease during pregnancy was exclusively observed in primiparous women. Both, positive coping strategies and higher perceived and received social support were independently associated with lower perceived stress, while evasive coping strategies were associated with higher levels of perceived stress. CONCLUSION: Our study reveals stress perception trajectories during pregnancies in primi- and multiparous women. Our findings underscore the need for intervention strategies aiming to improve social support and positive coping strategies especially in multiparous women in order to reduce the risks for adverse pregnancy outcomes.
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Adaptação Psicológica , Complicações na Gravidez/psicologia , Adulto , Estudos de Coortes , Feminino , Humanos , Estudos Longitudinais , Gravidez , Resultado da Gravidez , Estudos Prospectivos , Apoio Social , Estresse Psicológico , Inquéritos e QuestionáriosRESUMO
It has long been suspected that stress can cause hair loss, although convincing evidence of this has been unavailable. Here, we show that in mice sonic stress significantly increased the number of hair follicles containing apoptotic cells and inhibited intrafollicular keratinocyte proliferation in situ. Sonic stress also significantly increased the number of activated perifollicular macrophage clusters and the number of degranulated mast cells, whereas it down-regulated the number of intraepithelial gd T lymphocytes. These stress-induced immune changes could be mimicked by injection of the neuropeptide substance P in nonstressed mice and were abrogated by a selective substance P receptor antagonist in stressed mice. We conclude that stress can indeed inhibit hair growth in vivo, probably via a substance P-dependent activation of macrophages and/or mast cells in the context of a brain-hair follicle axis.
Assuntos
Encéfalo/fisiologia , Folículo Piloso/crescimento & desenvolvimento , Estimulação Acústica , Animais , Apoptose/efeitos dos fármacos , Degranulação Celular , Divisão Celular/efeitos dos fármacos , Citocinas/biossíntese , Folículo Piloso/química , Folículo Piloso/efeitos dos fármacos , Antígenos de Histocompatibilidade Classe II/biossíntese , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Indóis/farmacologia , Isoindóis , Queratinócitos/química , Queratinócitos/citologia , Queratinócitos/efeitos dos fármacos , Antígeno Ki-67/análise , Macrófagos/metabolismo , Macrófagos/patologia , Mastócitos/fisiologia , Camundongos , Receptores de Antígenos de Linfócitos T gama-delta/biossíntese , Pele/metabolismo , Pele/patologia , Estresse Fisiológico/fisiopatologia , Substância P/farmacologia , Linfócitos T/citologia , Linfócitos T/metabolismo , Regulação para CimaRESUMO
INTRODUCTION: The aim of this study was to develop an automatic differentiation of two perfusion compartments within the mouse placenta based on times of maximal contrast enhancement for a detailed and reproducible perfusion assessment. METHODS: Placentas (n = 17) from pregnant BALB/c mice (n = 10) were examined in vivo at 7T on gestation day 16.5. Coronal dual-echo 3D T1-weighted gradient-echo sequences were acquired after application of contrast agent for dynamic MRI. An adapted gamma variate function was fitted to the discrete concentration time curves to evaluate the effect of noise on perfusion and segmentation results. Time-to-peak maps based on fitted and discrete curves of each placenta were used to classify each voxel into the high- or low-blood flow compartment using k-means clustering. Perfusion analysis was performed using the steepest slope model and also applied to fitted and discrete curves. Results were compared to manually defined compartments from two independent observers using the Dice coefficient D. RESULTS: Manually defined placental areas of high-flow and low-flow were similar to the automatic segmentation for discrete (D = 0.76/0.75; D = 0.76/0.79) and fitted (D = 0.80/0.80; D = 0.81/0.82) concentration time curves. Mean perfusion values of discrete and fitted curves ranged in the high-flow compartment from 134 to 142 ml/min/100 ml (discrete) vs. 138-143 ml/min/100 ml (fitted) and in the low-flow compartment from 91 to 94 ml/min/100 ml (discrete) vs. 74-82 ml/min/100 ml (fitted). DISCUSSION: Our novel approach allows the automatic differentiation of perfusion compartments of the mouse placenta. The approach may overcome limitations of placental perfusion analyses caused by tissue heterogeneity and a potentially biased selection of regions of interest.
Assuntos
Modelos Cardiovasculares , Circulação Placentária , Placentação , Algoritmos , Animais , Velocidade do Fluxo Sanguíneo , Análise por Conglomerados , Meios de Contraste , Imagem Ecoplanar , Feminino , Hibridização Genética , Interpretação de Imagem Assistida por Computador , Imageamento Tridimensional , Masculino , Meglumina/análogos & derivados , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos DBA , Compostos Organometálicos , Perfusão , Gravidez , Razão Sinal-RuídoRESUMO
The notion that patients with eating disorders maintain a functional immunosurveillance in spite of severe malnutrition has attracted researchers for years. Dipeptidyl Peptidase IV (DPP IV), a serine protease with broad tissue distribution and known activity in serum, operates in the cascade of immune responses. Membrane-bound DPP IV expressed on lymphocytes, also known as the leukocyte antigen CD26, is considered to participate in T cell activation. We hypothesized that the activity of DPP IV in serum and expression of CD26 in lymphocytes may be altered in patients with eating disorders. Serum DPP IV activity and the number of CD26 (DPP IV)-positive peripheral blood lymphocytes were measured in 44 patients (anorexia nervosa (AN): n = 21, bulimia (B): n = 23) in four consecutive weekly analyses. The analysis of CD26-positive cells included the characterization of CD26-bright and CD26-dim positive subsets. Additionally, the expression of CD25 (IL-2 Receptor alpha chain) was evaluated to estimate the degree of T cell activation. The same analyses were carried out in healthy female volunteers (HC, n = 20). CD26-positive cells were reduced in patients as compared to healthy controls (mean 40.2% (AN) and 41.1% (B) vs. 47.4% (HC), p < 0.01), while the DPP IV activity in serum was elevated (mean 108.4 U/l (AN) and 91.1 U/l (B) vs. 80.3 U/l (HC), p < 0.01). The potential implications of changes in DPP IV expression and serum activity on--and beyond--immune function are discussed.
Assuntos
Anorexia Nervosa/enzimologia , Bulimia/enzimologia , Dipeptidil Peptidase 4/sangue , Subpopulações de Linfócitos/enzimologia , Adulto , Anorexia Nervosa/imunologia , Índice de Massa Corporal , Bulimia/imunologia , Estudos Transversais , Feminino , Humanos , Imunocompetência , Imunofenotipagem , Contagem de Linfócitos , Receptores de Interleucina-2/biossíntese , Receptores de Interleucina-2/genéticaRESUMO
Spontaneous abortion is the most common adverse pregnancy outcome, and stress has been suggested to be abortogenic in mice and humans. Stress-and/or pregnancy-related hormones (corticotropin releasing hormone, adrenocorticotropin, and progesterone) might interact with peripheral and local immunocompetent cells, such as certain T cell, mast cells or NK cells, and result in changes of cytokine production. In an established murine model, abortions can be triggered by exposing the mice to stress during early gestation. Recent data from this model indicated that increased levels of abortogenic Th1 cytokines, a decrease of progesterone and thus, PIBP were incongruous with successful pregnancy maintenance. Supplementation of progesterone exerts a pregnancy protective effect by induction of a pregnancy-protective Th2 biased immune response. Interestingly, data from a prospective study on human pregnancy revealed that women with a clinically normally progressing pregnancy but low levels of progesterone during the first trimester eventually suffered from a miscarriage. These data indicate that stress may lead to increase abortions by altering the endocrine system, which triggers an immune bias towards an abortogenic cytokine profile. Progesterone may be a good marker to identify a putative thread of a miscarriage in human and progesterone replacement therapy may abrogate this thread by inducing a Th2 biased immune response from the decidua.
Assuntos
Aborto Espontâneo/psicologia , Implantação do Embrião/imunologia , Estresse Psicológico/complicações , Aborto Espontâneo/imunologia , Animais , Citocinas/biossíntese , Implantação do Embrião/fisiologia , Feminino , Humanos , Camundongos , Modelos Animais , Gravidez , Proteínas da Gravidez/metabolismo , Progesterona/sangue , Estresse Psicológico/metabolismo , Fatores Supressores Imunológicos/metabolismoRESUMO
Prenatal stress (PS) is a known risk factor for several psychiatric diagnoses, including schizophrenia, attention deficit hyperactivity disorder, autism, anxiety, and depression which have been associated with serotonin transporter (SERT) dysregulation. Moreover, long-term effects in animal models associate with higher levels of immediate early genes, e.g. c-FOS (up-regulated in response to neuronal activity), in the brain of PS offspring. We therefore quantified the expression of both protein related mRNAs in adolescent BALB/c mice subjected to mild auditory stress on two separate days in mid gestation. SERT and c-FOS consistently correlated in most brain regions of PS mice and controls. Moreover, two-way ANOVAs revealed concomitantly increased levels of proteins, as well as of FOSL1 and FOSL2 mRNA, especially in the striatum and hippocampus of the PS offspring. Sex affected only and less consistently mRNA expression, yet interacted with PS, demonstrating that glucocorticoid receptor mRNA expression decreased in PS males but increased in PS females compared to the respective controls. This first finding of a correlation between SERT and c-FOS protein expression affected by PS, together with related mRNAs, may be considered a new target for behavioral and treatment studies in offspring.
Assuntos
Corpo Estriado/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Hipocampo/metabolismo , Efeitos Tardios da Exposição Pré-Natal/patologia , Proteínas Proto-Oncogênicas c-fos/metabolismo , Proteínas da Membrana Plasmática de Transporte de Serotonina/metabolismo , Análise de Variância , Animais , Animais Recém-Nascidos , Feminino , Antígeno 2 Relacionado a Fos/genética , Antígeno 2 Relacionado a Fos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Gravidez , Efeitos Tardios da Exposição Pré-Natal/etiologia , Proteínas Proto-Oncogênicas c-fos/genética , RNA Mensageiro/metabolismo , Proteínas da Membrana Plasmática de Transporte de Serotonina/genética , Caracteres Sexuais , Estresse Psicológico/complicaçõesRESUMO
Atherosclerosis is the underlying cause of cardiovascular disease and stroke. Endothelial cell dysfunctions are early events in atherosclerosis, resulting in the recruitment of circulating monocytes. The immune system can elicit an inflammatory response toward the atherosclerotic lesion, thereby accelerating lesion growth. Risk factors for atherosclerosis include hypertension, smoking, stress perception or low birth weight. As prenatal stress challenge decreases the birth weight and affects the offspring's postnatal immune response, we aimed to investigate whether prenatal stress contributes to the development of atherosclerosis in mice. Syngenic pregnant apolipoprotein E-deficient (apoE-/-) dams were exposed to sound stress on gestation days 12.5 and 14.5. The presence and size of atherosclerotic plaques in the offspring at the age of 15 weeks was evaluated by histomorphology, accompanied by flow cytometric analysis of the frequency and phenotype of monocytes/macrophages and regulatory T (Treg) cells in the blood. Further, cytokine secretion of peripheral blood lymphocytes was analyzed. In response to prenatal stress challenge, an increased frequency of large atherosclerotic plaques was detectable in apoE-/- offspring, which was particularly profound in females. Prenatal stress also resulted in alterations of the offspring's immune response, such as a decreased frequency of Treg cells in blood, alterations of macrophage populations in blood and an increased secretion of inflammatory cytokines. We provide novel evidence that prenatally stressed adult offspring show an increased severity of atherosclerosis. As Treg cells are key players in dampening inflammation, the observed increase in atherosclerosis may be due to the lack of Treg cell frequency. Future interdisciplinary research is urgently required to understand the developmental origin of prenatal stress-induced atherosclerosis. The availability of our model may facilitate and foster such research endeavors.
Assuntos
Apolipoproteínas E/deficiência , Arterite/imunologia , Aterosclerose/imunologia , Efeitos Tardios da Exposição Pré-Natal/imunologia , Estresse Fisiológico/imunologia , Animais , Arterite/etiologia , Aterosclerose/complicações , Aterosclerose/patologia , Citocinas/sangue , Feminino , Citometria de Fluxo , Técnicas Histológicas , Leucócitos/imunologia , Camundongos , Camundongos Knockout , Gravidez , Som/efeitos adversosRESUMO
OBJECTIVES: The steepest slope model is a numerically robust and fast method for perfusion quantification. The purpose of this study was to evaluate if the steepest slope model can be used for quantifying placental perfusion in mice based on dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) datasets. MATERIAL AND METHODS: T1-weighted DCE MRI was performed in 5 pregnant BALB/c mice on gestation day (gd) 14.5 and in 5 mice on gd 16.5 using a 7T small animal MRI scanner. The placentas were manually delineated in the DCE datasets and the arterial input function (AIF) was selected from the kidney hilus. Placental perfusion was determined on a voxel-by-voxel basis using the steepest slope model. Perfusion was averaged over the entire placenta as well as separately calculated for the high-flow compartment within the central labyrinth zone and for the remaining low-flow placenta tissue. The AIF selection was independently performed by two observers for assessment of inter-observer differences. RESULTS: Mean perfusion on gd 14.5 was 135 ml/min/100 ml (standard deviation SD: 29 ml/min/100 ml placenta) and 112 ml/min/100 ml on gd 16.5 for the whole placenta (SD: 32 ml/min/100 ml). Perfusion in the high flow compartment in the central labyrinth was significantly higher (p ≤ 0.002) than in the low-flow compartment including the remaining placenta tissue: 184 ml/min/100 ml (SD: 39 ml/min/100 ml) vs. 119 ml/min/100 ml (SD 28 ml/min/100 ml) on gd 14.5 and 158 ml/min/100 ml (SD: 58 ml/min/100 ml) vs. 114 ml/min/100 ml (SD: 52 ml/min/100 ml of placenta) on gd 16.5. The mean relative inter-rater observer difference was 6%. CONCLUSION: The steepest slope model is a computationally simple method, which allows perfusion quantification in the mouse placenta. Furthermore, the results of this work indicate that the different placental compartments should be analyzed separately to prevent biased results due to averaging.
Assuntos
Imageamento por Ressonância Magnética/métodos , Placenta/irrigação sanguínea , Circulação Placentária , Animais , Meios de Contraste , Feminino , Interpretação de Imagem Assistida por Computador/métodos , Camundongos , Camundongos Endogâmicos BALB C , GravidezRESUMO
Sound stress exposure increases fetal loss via inflammatory pathways. Inflammation is known to up-regulate cell adhesion molecules, such as vascular cell adhesion molecule-1 (VCAM-1), which mediates the adhesion of leukocytes to the vascular endothelium. In this work, we studied the frequency of VCAM-1(+) vessels at the fetomaternal interface in stressed and non-stressed pregnant CBA/J female mice mated with DBA/2J (high fetal loss model) or BALB/c (low fetal loss model) males. The high fetal loss model had fewer large vessels on gestation day 6.5, and stress reduced the frequency of large vessels to a similar number in both high and low fetal loss models. In the high fetal loss model, however, the frequency of VCAM-1+ vessels was dramatically increased. This study shows that VCAM-1 expression is modulated by stress at the fetomaternal interface in abortion-prone cross-breeding.
Assuntos
Aborto Espontâneo/metabolismo , Regulação da Expressão Gênica , Placenta/metabolismo , Estresse Fisiológico , Molécula 1 de Adesão de Célula Vascular/biossíntese , Aborto Espontâneo/patologia , Animais , Feminino , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos CBA , Placenta/patologia , GravidezRESUMO
Controlled trophoblast invasion is a key process during human placentation and a prerequisite for successful pregnancy. Progesterone is one of the factors to regulate trophoblast invasiveness. Progesterone-induced blocking factor (PIBF) is a progesterone-induced molecule expressed by the trophoblast, and also by tumors. The distribution of PIBF within the first-trimester decidua coincides with sites of trophoblast invasion. Another molecule that has been implicated in the control of trophoblast invasiveness is placental leptin. Leptin inhibits the secretion of progesterone by cytotrophoblast. The aim of this work was to investigate the possible interaction of PIBF and leptins in regulating trophoblast invasion. Paraffin-embedded sections from normal first-trimester placentae, partial moles, complete moles, and choriocarcinomas were reacted with PIBF, leptin, and leptin receptor specific antibodies. PIBF-deficient trophoblast cells were generated using siRNA and leptin receptor was detected on Western blot analysis. The lysates of PIBF-treated cells were used for detecting leptin expression in a protein array. PIBF was expressed in both normal first-trimester villous trophoblast and in partial mole. Compared with this, PIBF expression was markedly decreased in complete mole and absent in choriocarcinoma. Neither leptinR nor leptin were detected in partial mole, whereas both of these molecules were present in complete mole and choriocarcinoma. Leptin receptor expression was upregulated in PIBF-deficient cells, while leptin expression was decreased in PIBF-treated cells. These data suggest that PIBF affects the expression of leptin and its receptor, and that PIBF expression is inversely related to trophoblast invasiveness.