In Vivo Flow Measurements of Murine Renal Arteries and Veins with High Frequency Ultrasound

The number of glomeruli in the kidneys has been shown to have an effect on the decline in renal function over time (Brenner, Garcia, Anderson 1988). Furthermore, flow in the renal arteries and veins may depend on the number of glomeruli in the kidney. Consistent in vivo measurements of volumetric flow in the renal arteries and veins are difficult to obtain. Thus, the purpose of this study was to develop non-invasive imaging techniques capable of estimating arterial and venous flow to kidneys. A high-frequency small animal ultrasound system was chosen based upon its excellent spatial and temporal resolution when imaging mice (Vevo 2100, VisualSonics, Inc.). Velocity profiles of the renal arteries and veins in C57BL/6 male mice (n=4) were measured. Motion, color Doppler, and pulsed wave Doppler data were acquired and used to determine renal diameter, maximum velocity, mean velocity, and volumetric flow for both kidneys. For the renal artery the average volumetric flow was 33.31±7.16 mm3/s and for the renal vein it was 30.23±4.58 mm3/s. The next step will be imaging the same animals multiple times to ensure that these measurements are consistent over prolonged periods of time. Then data will be collected from different breeds of mice to conclude whether or not differences in glomeruli number affect renal flow. Measurement of volumetric flow in the renal arteries and veins can lead to important insights into how the glomeruli density in kidneys relates to renal flow and function

Visualization of Complex Flow Patterns in Angiotensin II-Induced Dissecting Murine Abdominal Aortic Aneurysms with High Frequency Ultrasound

Abdominal aortic aneurysm (AAA) rupture is a common cause of mortality in the United States. Current treatments are only employed once the risk of rupture outweighs the risks associated with surgery. Murine models have been developed to characterize AAA pathogenesis in the hope that new treatments will be developed. For this study, angiotensin II (AngII) was infused subcutaneously into apolipoprotein E-deficient (ApoE-/-) mice using an osmotic mini-pump over 28 days. ApoE-/- mice (16-week-old, 3 females, 2 males) were imaged using a VisualSonics Vevo 2100 high frequency ultrasound before pump implantation and 3, 7, 14, 21, and 27 days following implantation. Images were acquired in the transverse and longitudinal planes from the suprarenal region of the aorta. Blood pressure measurements were taken using a tail-cuff system (CODA, Kent Scientific). Three mice (1 female, 2 male) developed aneurysms within the first 14 days of infusion. Pre-study abdominal aortas had a diastolic diameter of 0.84±0.09 mm and a systolic diameter of 0.96±0.08 mm. By day 21, AAAs had a diastolic diameter of 1.51±0.59 mm and a systolic diameter of 1.56±0.59 mm. Initially, mice had a systolic blood pressure of 111.94±6.53 mmHg and a diastolic pressure of 82.38±5.13 mmHg. These pressures steadily elevated but eventually began to plateau. By day 27, systolic pressure had risen to 154.92±11.43 mmHg and diastolic pressure to 115.77±10.25 mmHg. Color Doppler images revealed complex, recirculating flow within the aneurysms, a phenomenon which could affect vessel remodeling. In conclusion, this study utilized in vivo sonographic methods to characterize AAA development

Three Dimensional Quantification of Angiotensin II-Induced Murine Abdominal Aortic Aneurysms Using High Frequency Ultrasound

Abdominal aortic aneurysms (AAAs), a localized dilation of the vessel wall of 50% or more above normal, claims approximately 14,000 U.S. lives yearly due to aortic rupture. This commonly asymptomatic disease can only be treated by endovascular stent grafts or invasive surgery, usually after the AAA diameter reaches 5 cm. Because these treatment methods carry serious risk, stem cell therapy is being explored in order to provide a low risk option for managing smaller AAAs. To determine if stem cell therapy, once administered, could stabilize or reduce AAA growth, baseline 3D ultrasound measurements in a control group were first needed. High frequency ultrasound was used on apolipoprotein E-deficient (apoE-/-) mice given angiotensin II (AngII) from subcutaneously implanted osmotic mini pumps. This mouse model developed dissecting AAAs, containing a false and true lumen, which were clearly visualized and quantified using 3D ultrasound imaging. With this ultrasound technique, we found that aneurysm diameter, total volume, and false lumen volume all increased steadily over a period of 28 days once AAAs formed. These data suggest our noninvasive, 3D ultrasound technique can be used to monitor the progression of aneurysms that may be delayed once stem cell therapy is administered

Nonlinear Optical Microscopy of Murine Abdominal Aortic Aneurysm

Abdominal aortic aneurysm (AAA) is a cardiovascular disease characterized by dilation and weakening of the vessel wall. AAA rupture is responsible for approximately 14,000 deaths annually in the United States [1]. Nonlinear optical (NLO) microscopy presents new possibilities for analyzing AAA tissue samples from murine models. Common NLO techniques are two-photon excitation fluorescence (TPEF), which detects the intrinsic autofluorescent properties of elastin, and second-harmonic generation (SHG), which is specific for collagen fibrils. Elastin and collagen, two major extracellular matrix components, help the aortic wall withstand internal pressure. Murine AAAs were created through 1) subcutaneous continuous systemic infusion of angiotensin II (AngII) in hyperlipidemic apolipoprotein E-deficient mice and 2) by intraluminal infusion of elastase (low 0.5 U/ml and high 25 U/ml concentrations) into the infrarenal aorta of rats [2]. We imaged aneurysmal and control tissue using TPEF and SHG and compared the resulting images to sections stained with standard elastin and collagen markers. TPEF images revealed disorganized elastin sheets and SHG images indicated collagen turnover after aneurysm formation. We quantified the relative degree of elastin degradation and collagen content in the aortic media within a user-defined area on sections stained with Verhoeff-van Gieson (VVG) or Masson’s trichrome (MTC), as well as on TPEF and SHG images. Our analysis with VVG-stained sections shows that elastin content in AAA tissue is significantly decreased by 64% in AngII models (P=0.02), by 34% in low concentration elastase models (P=0.07), and by 99% in high concentration elastase models (P=0.03), relative to control aortic tissue

CB1 Expression Is Attenuated in Fallopian Tube and Decidua of Women with Ectopic Pregnancy

BACKGROUND: Embryo retention in the Fallopian tube (FT) is thought to lead to ectopic pregnancy (EP), a considerable cause of morbidity. In mice, genetic/pharmacological silencing of cannabinoid receptor Cnr1, encoding CB1, causes retention of embryos in the oviduct. The role of the endocannabinoids in tubal implantation in humans is not known. METHODS AND FINDINGS: Timed FT biopsies (n = 18) were collected from women undergoing gynecological procedures for benign conditions. Endometrial biopsies and whole blood were collected from women undergoing surgery for EP (n = 11); management of miscarriage (n = 6), and termination of pregnancy (n = 8). Using RT-PCR and immunohistochemistry, CB1 mRNA and protein expression levels/patterns were examined in FT and endometrial biopsies. The distribution of two polymorphisms of CNR1 was examined by TaqMan analysis of genomic DNA from the whole blood samples. In normal FT, CB1 mRNA was higher in luteal compared to follicular-phase (p<0.05). CB1 protein was located in smooth muscle of the wall and of endothelial vessels, and luminal epithelium of FT. In FT from women with EP, CB1 mRNA expression was low. CB1 mRNA expression was also significantly lower (p<0.05) in endometrium of women with EP compared to intrauterine pregnancies (IUP). Although of 1359G/A (rs1049353) polymorphisms of CNR1 gene suggests differential distribution of genotypes between the small, available cohorts of women with EP and those with IUP, results were not statistically significant. CONCLUSIONS: CB1 mRNA shows temporal variation in expression in human FT, likely regulated by progesterone. CB1 mRNA is expressed in low levels in both the FT and endometrium of women with EP. We propose that aberrant endocannabinoid-signaling in human FT leads to EP. Furthermore, our finding of reduced mRNA expression along with a possible association between polymorphism genotypes of the CNR1 gene and EP, suggests a possible genetic predisposition to EP that warrants replication in a larger sample pool

Ectopic Pregnancy as a Model to Identify Endometrial Genes and Signaling Pathways Important in Decidualization and Regulated by Local Trophoblast

The endometrium in early pregnancy undergoes decidualization and functional changes induced by local trophoblast, which are not fully understood. We hypothesized that endometrium from tubal ectopic pregnancy (EP) could be interrogated to identify novel genes and pathways involved in these processes. Gestation-matched endometrium was collected from women with EP (n = 11) and intrauterine pregnancies (IUP) (n = 13). RNA was extracted from the tissue. In addition, tissues were prepared for histological analysis for degree of decidualization. We compared a) the samples from EP that were decidualized (n = 6) with non-decidualized samples (n = 5), and b) the decidualized EP (n = 6) with decidualization-matched IUP (n = 6) samples using an Affymetrix gene array platform, with Ingenuity Pathway Analysis, combined with quantitative RT-PCR. Expression of PRL and IGFBP1 was used to confirm the degree of decidualization in each group. There were no differences in PRL or IGFBP1 expression in the decidualization-matched samples but a marked reduction (P<0.001) in the non-decidualized samples. Decidualization was associated with increased expression of 428 genes including SCARA5 (181-fold), DKK1 (71-fold) and PROK1 (32-fold), and decreased expression of 230 genes including MMP-7 (35-fold) and SFRP4 (21-fold). The top canonical pathways associated with these differentially expressed genes were Natural Killer Cell and Wnt/b-Catenin signaling. Local trophoblast was associated with much less alteration of endometrial gene expression with an increase in 56 genes, including CSH1 (8-fold), and a reduction in 29 genes including CRISP3 (8-fold). The top associated canonical pathway was Antigen Presentation. The study of endometrium from tubal EP may promote novel insights into genes involved in decidualization and those influenced by factors from neighboring trophoblast. This has afforded unique information not highlighted by previous studies and adds to our understanding of the endometrium in early pregnancy

Weight Strain And Binge Drinking Among Adolescents

Obesity and substance use are two common areas of research among adolescents. Interestingly, very little research examines the relationship between these two important health risk behaviors and the findings are inconsistent. Guided by Agnew\u27s general strain theory and using the Add Health data, we examine this neglected area of research. The current research has identified a link between weight strain and binge drinking and is supportive of the extant research on both general strain theory and the links between stigma, stress, and health. We also found some evidence that this relationship was gendered. Implications and future research directions are discussed. © Taylor & Francis Group, LLC

Influence of life course socioeconomic position on older women's health behaviors: findings from the British Women's Heart and Health Study.

OBJECTIVES: We examined the association between health behaviors and socioeconomic status (SES) in childhood and adult life. METHODS: Self-reported diet, smoking, and physical activity were determined among 3523 women aged 60 to 79 years recruited from general practices in 23 British towns from 1999 through 2001. RESULTS: The most affluent women reported eating more fruit, vegetables, chicken, and fish and less red or processed meat than did less affluent women. Affluent women were less likely to smoke and more likely to exercise. Life course SES did not influence the types of fat, bread, and milk consumed. Adult SES predicted consumption of all foods considered and predicted smoking and physical activity habits independently of childhood SES. Childhood SES predicted fruit and vegetable consumption independently of adult SES and, to a lesser extent, predicted physical activity. Downward social mobility over the life course was associated with poorer diets and reduced physical activity. CONCLUSIONS: Among older women, healthful eating and physical activity were associated with both current and childhood SES. Interventions designed to improve social inequalities in health behaviors should be applied during both childhood and adult life

Development of Non-Invasive In Vivo Ultrasound Imaging Techniques for Elastase-Induced Experimental Abdominal Aortic Aneurysms

Abdominal aortic aneurysms (AAAs) are pathological dilations of the aorta which are associated with significant morbidity and mortality. The underlying mechanisms that cause this inflammatory disease are not fully understood and thus, are currently under investigation. In the hopes of preventing disease progression, rodent models that mimic the human condition have been developed to provide insight into the pathogenesis of AAAs. In this study, porcine pancreatic elastase (0.44 U; Sigma-Aldrich) was infused into the infrarenal aortas of male, Sprague Dawley rats to induce aneurysms. To perform the surgery, temporary ligatures were placed around proximal and distal sections of the abdominal aorta and a catheter was inserted into the vessel through an aortotomy slightly above the trifurcation to infuse elastase (30 minutes). Rats were imaged using a VisualSonics Vevo 2100 high-frequency ultrasound prior to and following surgery on days 3, 7, 14, 21, and 28. Of the 8 rats used in this study, 4 survived for 28 days and developed aneurysms. Pre-surgery abdominal aortas had a systolic diameter of 1.16 ± 0.19 mm and a diastolic diameter of 0.99 ± 0.17 mm. By day 14, AAAs had a systolic diameter of 2.32 ± 0.63 mm and a diastolic diameter of 2.25 ± 0.64 mm. Our efforts using this rat model will benefit future mesenchymal stem cell work aimed at preventing aneurysm formation by modulating the immune response. In conclusion, this study utilized high-frequency ultrasound to characterize an elastase-induced rat AAA model and will help increase our understanding of aneurysm pathogenesis