Gland Fat Research Articles

EGCG, a major green tea catechin suppresses breast tumor angiogenesis and growth via inhibiting the activation of HIF-1α and NFκB, and VEGF expression

The role of EGCG, a major green tea catechin in breast cancer therapy is poorly understood. The present study tests the hypothesis that EGCG can inhibit the activation of HIF-1α and NFκB, and VEGF expression, thereby suppressing tumor angiogenesis and breast cancer progression. Sixteen eight-wk-old female mice (C57BL/6 J) were inoculated with 10^6 E0771 (mouse breast cancer) cells in the left fourth mammary gland fat pad. Eight mice received EGCG at 50–100 mg/kg/d in drinking water for 4 weeks. 8 control mice received drinking water only. Tumor size was monitored using dial calipers. At the end of the experiment, blood samples, tumors, heart and limb muscles were collected for measuring VEGF expression using ELISA and capillary density (CD) using CD31 immunohistochemistry. EGCG treatment significantly reduced tumor weight over the control (0.37 ± 0.15 vs. 1.16 ± 0.30 g; P < 0.01), tumor CD (109 ± 20 vs. 156 ± 12 capillary #/mm^2; P < 0.01), tumor VEGF expression (45.72 ± 1.4 vs. 59.03 ± 3.8 pg/mg; P < 0.01), respectively. But, it has no effects on the body weight, heart weight, angiogenesis and VEGF expression in the heart and skeletal muscle of mice. EGCG at 50 μg/ml significantly inhibited the activation of HIF-1α and NFκB as well as VEGF expression in cultured E0771 cells, compared to the control, respectively. These findings support the hypothesis that EGCG, a major green tea catechin, directly targets both tumor cells and tumor vasculature, thereby inhibiting tumor growth, proliferation, migration, and angiogenesis of breast cancer, which is mediated by the inhibition of HIF-1α and NFκB activation as well as VEGF expression.

2-NBDG Fluorescence Imaging of Hypermetabolic Circulating Tumor Cells in Mouse Xenograft model of Breast Cancer

To determine use of 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG) as a tracer for detection of hypermetabolic circulating tumor cells (CTC) by fluorescence imaging. Human breast cancer cells were implanted in the mammary gland fat pad of athymic mice to establish orthotopic human breast cancer xenografts as a mouse model of circulating breast cancer cells. Near-infrared fluorescence imaging of the tumor-bearing mice injected with 2-DeoxyGlucosone 750 (2-DG 750) was conducted to assess glucose metabolism of xenograft tumors. Following incubation with fluorescent 2-NBDG, circulating breast cancer cells in the blood samples collected from the tumor-bearing mice were collected by magnetic separation, followed by fluorescence imaging for 2-NBDG uptake by circulating breast cancer cells, and correlation of the number of hypermetabolic circulating breast cancer cells with tumor size at the time when the blood samples were collected. Human breast cancer xenograft tumors derived from MDA-MB-231, BT474, or SKBR-3 cells were visualized on near-infrared fluorescence imaging of the tumor-bearing mice injected with 2-DG 750. Hypermetabolic circulating breast cancer cells with increased uptake of fluorescent 2-NBDG were detected in the blood samples from tumor-bearing mice and visualized by fluorescence imaging, but not in the blood samples from normal control mice. The number of hypermetabolic circulating breast cancer cells increased along with growth of xenograft tumors, with the number of hypermetabolic circulating breast cancer cells detected in the mice bearing MDA-MB231 xenografts larger than those in the mice bearing BT474 or SKBR-3 xenograft tumors. Circulating breast cancer cells with increased uptake of fluorescent 2-NBDG were detected in mice bearing human breast cancer xenograft tumors by fluorescence imaging, suggesting clinical use of 2-NBDG as a tracer for fluorescence imaging of hypermetabolic circulating breast cancer cells.

Neuropathies amyloïdes héréditaires : aspects cliniques et neuropathologiques

Transthyretin familial amyloid polyneuropathy (TTRFAP) is an autosomal dominant neuropathy that is fatal within about 10 years after symptom onset. TTRFAP is observed worldwide, albeit with a higher frequency of the most common variant, Val30met, in Portugal, Sweden and Japan. Various phenotypic differences are observed. TTRFAP should be considered in patients with a progressive axonal polyneuropathy of unknown origin, especially when associated with autonomic nervous system dysfunction. A positive family history is found in most cases when onset begins around 30 years of age, while late-onset FAP is often sporadic and may be confused with chronic inflammatory demyelinating polyneuropathy. Nerve biopsy is often used to confirm the presence of extracellular amyloid deposits in interstitial tissue of the endoneurial space, although amyloid can also befound in muscle, salivary gland and abdominal fat. It is important to stress that biopsy negativity does not rule out amyloidosis. Genetic testing for TTR gene mutations should be performed in case of progressive length-dependent axonal polyneuropathy predominantly involving small nerve fibers.

Altered regional blood flow distribution in Mas-deficient mice

We have recently shown that the acute infusion of angiotensin-(1-7) [Ang-(1-7)] or chronic increase in plasma Ang-(1-7) produces important changes in regional blood flow in rats. To further assess whether these changes are related to Mas, in this study hemodynamic measurements were performed in Ang-(1-7) receptor Mas knockout C57BL/6 (Mas-KO) mice and age-matched wild type (WT) control mice, using fluorescent microspheres. Mean arterial pressure in urethane-anesthetized Mas-KO mice (12-16 weeks old) did not differ from that in WT mice (79 ± 2 and 80 ± 2 mmHg respectively). However, pronounced differences were observed in other hemodynamic measurements. Mas-KO mice exhibited a significant decrease in stroke volume (0.03 ± 0.01 versus 0.05 ± 0.01 ml/beat in WT) and decreased cardiac index (0.81 ± 0.08 versus 1.24 ± 0.24 ml/min/g in WT). Strikingly, Mas-KO mice exhibited a marked increase in vascular resistance and a decrease in blood flow in the kidney, lung, adrenal gland, mesentery, spleen and brown fat tissue. The decrease in blood flow ranged from 34% (spleen) to 55% (brown fat tissue). These results suggest that the Ang-(1-7)/Mas axis plays an important role in regional and systemic hemodynamic adjustments in mice.

Cutaneous haemorrhage induced by minimal trauma as a sign of light chain‐associated amyloidosis

An 82-year-old patient was hospitalized with a deep dissecting haematoma of the leg. There had been no preceding trauma. She had a previous history of high blood pressure, atrial fibrillation, gastro-intestinal bleeding and coronary heart disease. Her medications included lysine acetylsalicylate 75 mg/day. Physical examination showed pitting oedema of both legs and bilateral decreased vesicular breath sounds. She had periorbital purpura, petechiae on the hands, and distal onycholysis. Petechiae were subsequently noticed in areas submitted to microtrauma (plaster, catheter). Firm friction on the skin with the edge of a pen (left) induced linear petechiae (right). Investigations revealed anaemia (haemoglobin concentration 91 g/l), leucocytosis (white blood cell count 26·7 × 109/l) and increased C-reactive protein (126 mg/l). Renal function was normal. The patient had hypogammaglobulinemia (4·6 g/l), an increase in serum lambda free light chains (331 mg/l) and lambda Bence–Jones proteinuria. Bone marrow plasma cells were 8%, some of which were cytologically abnormal. Computed tomography scanning showed bilateral pleural effusions. Echocardiography was abnormal, suggesting hypertrophic cardiomyopathy. A skeletal survey showed no lytic lesions. Amyloid fibrils were identified on salivary gland and abdominal fat biopsies. A diagnosis of systemic AL amyloidosis was thus made. The patient was treated with melphalan, dexamethasone and intravenous immunoglobulin therapy but she nevertheless died of cardiac failure 6 months after the start of the treatment. The use of local trauma to induce intralesional haemorrhage in cutaneous lesions has previously been described as a clinical test of value in the diagnosis of systemic amyloidosis (Hurley & Weinberg, 1964). We report petechiae induced by minimal trauma in apparently normal skin as a clinical sign that is helpful in the diagnosis of primary amyloidosis and is easy to induce without the need for any specific equipment. Because of amyloid fibrils in capillary walls, mild trauma can cause the rupture of capillaries, even in apparently normal skin. Linear skin friction can result in linear petechiae or purpura; this observation is an indication for further investigation, in order to confirm a diagnosis of systemic amyloidosis. Purpura caused by capillary fragility is not specific for amyloidosis. It can be seen also in chronic renal failure, liver cirrhosis, skin ageing, long-standing corticosteroid therapy and in primary haemostatic disorders. We hypothesize that the dissecting haematoma was also a result of amyloidosis-related capillary fragility, as the patient did not have any other predisposing factor.

A Study on the Effect of High Calorie Diet on Stress Alleviation

In this study, we tried to reveal the presence of a cytokine that is regulated by weight, food intake, body fat, the hypothalamus-pituitary-adrenal axis, and corticosterone. The experiment group was given a high calorie diet and the control group was fed a normal calorie diet, and both groups were exposed to various stresses. The behavioral and hormonal change by the kinds and severity of stresses were measured via their immunologic aspect as well as their stress alleviation by voluntary high calorie food intake. After sacrificing the animal, the adrenal gland fat tissues were weighed, and the high calorie diet group showed heavier adrenal glands than those of the control group. In particular, the CVS A group had lighter adrenal gland weight than the Res B and CVS B groups, and the other groups had heavier adrenal glands. The comparison of plasma ACTH concentration to the corticosteroid secretion ratio showed that the CVS A group had decreased ACTH and an increased corticosteroid/ACTH ratio. This fact shows that the adrenal gland was highly active and, in spite of a normal corticosteroid concentration, induced too much negative feedback inhibition.

The intraductal injection of 5-fluorouracil for breast cancer: Preventive effect and immune response.

e13559 Background: Chemotherapy has been shown to act as an immune response modifier. Many groups are using chemotherapy to induce immune responses such as by using low doses of cyclophosphamide. Building up on our previous work (Murata et al., 2006, Stearns et al., 2011), we demonstrate that intraductal (i.duc), but not the intravenous (IV) route of injection of chemotherapy to HER2/neu transgenic mouse mammary glands (MMG) alters the tumor environment to effectively induce immune effector cells. Methods: 1.Tumor-free survival: 5-fluorouracil (5FU) was administered intraductally to the MMGs on the left side of mice; the MMG on right side were untreated. 2. FACS analysis: 20w mice were administered 5FU 2 times in a 4 week interval i.duc only to the left side, IV, or no treatment (NT). At week 5, the mice were sacrificed and the regional lymph nodes (RLN) and spleen were removed. Lymphocytes from RLN and spleen were analyzed. 3. Tumor re-challenge: GT8 cancer cells were injected into the one side of 4th mammary gland fat pad of 12 week mice. The mice were then injected i.duc with 5-FU to all teats in the tumor implanted side, or IV or given NT. The surviving mice were re-challenged with GT8 cancer cells into the ipsilateral 3rd mammary gland fat pad, 4 weeks after the first inoculation. Results: 1. The tumor incidence in the 5FU treated side after 16 weeks was significantly lower (13% of 70 glands) compared to mice that received NT (34% of 150, p=0.001), and IV (28% of 70, p=0.03). The tumor incidence in the untreated side of 5FU-treated mice was also significantly lower (18% of 70) compared to the NT (p=0.01). 2. The number of CD8 T cell showed no change among the groups in the RLN but was significantly lower (p=0.01) in the spleen of the i.duc. The number of CD62LLOW+ T cell in treated side of RLN was significantly higher compared to the IV (p=0.02) and NT (p=0.02). The number of CD62LLOW+ was low in the spleen. 3. The survivors in the group previously treated with i.duc 5-FU were able to achieve a lasting rejection of the tumor re-challenge when compared to mice who previously received 5-FU IV. Conclusions: I.duc administration of 5FU into MMG effectively induced immune effector cells and prevented mammary tumor growth in HER2/neu transgenic mice.

Antioxidant and Protective Effects ofBupleurum falcatumon the L-Thyroxine-Induced Hyperthyroidism in Rats

Bupleuri Radix (BR), the dried roots of Bupleurum falcatum L., has been used in folk medicine as an antiinflammatory and antioxidative agent. The aqueous extract of BR was evaluated for its possible ameliorative effect in the regulation of hyperthyroidism in l-thyroxine- (LT4-) induced rat model. After oral administration of 300, 150, and 75 mg/kg of BR extracts, once a day for 15 days from 12th LT4 treatments, changes on the body, thyroid gland, liver, and epididymal fat pad weights, serum triiodothyronine, thyroxine, thyroid-stimulating hormone, asparte aminotransferase and alanine aminotransferase concentrations, hepatic lipid peroxidation, glutathione contents, superoxide dismutase, and catalase activities were investigated with thyroid gland, liver, and epididymal fat histopathological changes. The effects of BR extracts were compared with that of propylthiouracil, a standard antithyroid drug 10 mg/kg (intraperitoneally). In this experiment, BR extracts dose dependently reversed LT4-induced hyperthyroidisms, and these effects indicating their potential in the regulation of hyperthyroidism. Further, the BR extract normalized LT4-induced liver oxidative stresses, and also reduced liver and epididymal fat pad changes. BR extracts 150 mg/kg showed comparable effects on the LT4-induced rat hyperthyroidism as compared with PTU 10 mg/kg. These effects of BR may help the improvement of hyperthyroidisms and accompanied various organ damages.

P3-17-07: EGCG, a Green Tea Antioxidant Suppresses Breast Tumor Angiogenesis and Growth Via Inhibiting the Activation of HIF-1a and NFκB, and VEGF Expression.

Abstract Background Cancer cells are under greater oxidative stress than normal cells. Limited studies have showed that epigallocatechin-3-gallate (EGCG), a green tea antioxidant can afford protection against a variety of cancer types. The role of EGCG in breast cancer therapy is poorly understood. The present study tests the hypothesis that EGCG as an antioxidant can inhibit the activation of HIF-1α and NFκB, and VEGF expression, thereby suppressing tumor angiogenesis and breast cancer progression. Material and Methods: 16 eight-wk-old female mice (C57BL/6J) were inoculated with 10^6 E0771 (mouse breast cancer) cells in the left fourth mammary gland fat pad. 8 mice received EGCG at 50–100 mg/kg/d in drinking water for 4 weeks. 8 control mice received drinking water only. Tumor size was monitored using dial calipers. At the end of the experiment, blood samples, tumors, heart and limb muscles were collected for measuring VEGF expression using ELISA and capillary density (CD) using CD31 immunochemistry. Cultured E0771 cells were used for determining the direct effects of EGCG on proliferation (3H-thymidine incorporation), migration (Matrigel assay), VEGF expression (ELISA), the activation of HIF-1α and NFκB (motif binding assays, Active Motif). MCF-7 and MDA-MB-231 cells were also used for 3H-thymidine incorporation. Results: Oral EGCG treatment significantly reduced tumor weight over the control (0.37±0.15 vs. 1.16±0.30 g; P&amp;lt;0.01), tumor CD (109±20 vs. 156±12 capillary #/mm^2; P&amp;lt;0.01), tumor VEGF expression (45.72±1.4 vs. 59.03±3.8 pg/mg; P&amp;lt;0.01), respectively. EGCG treatment reduced plasma VEGF levels over the control mice (26.48±3.76 vs. 40.79±3.5 pg/ml; P&amp;lt;0.01). However, there were no differences in the body weight and heart weight between EGCG and the control groups. EGCG did not inhibit angiogenesis and VEGF expression in the heart and skeletal muscle of mice, compared to the control. EGCG at 50 mmol/L significantly inhibited the activation of HIF-1α (0.11±0.02 vs. 0.24±0.02; P&amp;lt;0.01) and NFκB (1.15±0.21 vs. 1.61±0.32; P&amp;lt;0.01) as well as VEGF expression (1752±49 vs. 2254±91 pg/mg; P&amp;lt;0.01) in cultured E0771 cells, compared to the control, respectively. EGCG caused a dose-related inhibition on the proliferation and migration in cultured E0771 cells. EGCG also caused a dose-related inhibition on the proliferation in cultured MCF-7 and MDA-MB-231 cells. Discussion: These findings support the hypothesis that EGCG, a green tea antioxidant, directly targets both tumor cells and tumor vasculature, thereby inhibiting tumor growth, proliferation, migration, and angiogenesis of breast cancer, which is mediated by the inhibition of HIF-1α and NFκB activation as well as VEGF expression. Interestingly, oral EGCG treatment has no effects on the body weight, heart weight, angiogenesis and VEGF expression in the heart and skeletal muscle of mice. This work will have important implications for translating EGCG therapy to human breast cancer treatment and prevention. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-17-07.

Activation of the Aryl Hydrocarbon Receptor by TCDD Inhibits Mammary Tumor Metastasis in a Syngeneic Mouse Model of Breast Cancer

Treatment with aryl hydrocarbon receptor (AhR) agonists can slow or reverse the growth of primary mammary tumors in rodents, which has fostered interest in developing selective AhR modulators for treatment of breast cancer. However, the major goal of breast cancer therapy is to inhibit metastasis, the primary cause of mortality in women with this disease. Studies conducted using breast cancer cell lines have demonstrated that AhR agonists suppress proliferation, invasiveness, and colony formation in vitro; however, further exploration using in vivo models of metastasis is warranted. To test the effect of AhR activation on metastasis, 4T1.2 mammary tumor cells were injected into the mammary gland fat pad of syngeneic Balb/c mice treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Primary tumor growth was monitored for 4 weeks, at which time metastasis was determined. TCDD treatment suppressed metastasis by approximately 50%, as measured both in the lung and in mammary glands at sites distant from the primary tumor. Primary tumor growth was not suppressed by TCDD exposure nor was proliferation of 4T1.2 cells affected by TCDD treatment in vitro. Taken together, these results suggest that the protective effect of AhR activation was selective for the metastatic process and not simply the result of a direct decrease in tumor cell proliferation or survival at the primary site. These observations in immunologically intact animals warrant further investigation into the mechanism of the protective effects of AhR activation and support the promise for use of AhR modulators to treat breast cancer.

A New Technique of Upper Eyelid Blepharoplasty Using the Orbicularis Muscle Flap

Blepharoplasty of the upper eyelids is one of the most commonly performed procedures in aesthetic plastic surgery. The orbicularis muscle flap technique provides good results for patients with atonic skin and an atonic orbicularis muscle when there is excessive fullness in the lateral aspect of the upper lid. This flap also can be used to contain a prominent lacrimal gland or retro-orbicularis oculi fat (ROOF) without dangerous removal of the last one. A total of 30 patients underwent upper lid blepharoplasty through a supratarsal incision approach. Dissection of the orbicularis muscle flap was made 5-8mm above the supratarsal fold. Before the muscle flap suspension, the excess skin and fat pad were removed. The flap suspension was achieved with its rotation and motion of a tunnel in the suborbicularis plane, which anchored it on the periosteum of the orbital lateral rim of the frontal bone with a permanent suture. This technique restores the sharp and crisp tarsal folds and pretarsal show with deepening of the orbitopalpebral sulcus. The orbicularis muscle flap also can be proposed for cases with a ptosis of the lateral upper lid. The authors believe this technique reduces the risk for "rounding" of the eyelid and produces better contact of the lid with the ocular globe. The orbicularis muscle flap procedure is a feasible technique that can be used reliably to reshape the lateral upper lid. It would be a useful addition to the armamentarium of the plastic surgeon who is called on to perform upper blepharoplasty.

Abstract LB-456: The intraductal injection of 5-fluorouracil induces immune response to eradicate and prevent breast cancer

Abstract Chemotherapy has been shown to act as an immune response modifier. Many groups, incuding our own, are using chemotherapy to induce immune responses such as by using low doses of cyclophosphamide. Our previous work has demonstrated that anti-cancer agents can eliminate new/existing tumors with minimal toxicity in rat and mouse models of breast cancer. Here we demonstrate that intraductal (i.duc), but not the intravenous (IV) route of injection, of chemotherapy to mammary glands alters the tumor environment to effectively induce immune effector cells. We used the mouse HER2/neu (neu/N) transgenic, spontaneous mammary tumor model. 5-fluorouracil (5FU) was administered intraductally to the mammary glands on the left side of parous mice whereas the mammary glands on right side received no treatment (NT). The mammary tumor incidence in the 5FU treated side was significantly lower compared to mice that received NT, and IV treated group. Interestingly the incidence of mammary tumors in the untreated side of 5FU-treated mice was also significantly lower compared to the NT and IV group. We hypothesized that the protection afforded to the contralateral chain of mammary glands by ipsilateral i.duc administration of 5FU, may be mediated through an immunological mechanism. Twenty week parous mice were administered 5FU either through i.duc injection, only to left side teats, IV injection, or NT. The mice received 5FU 2 times in a 4 week interval. A week after the second treatment, the mice were sacrificed and the regional lymphnodes (RLN) of the 3rd and 4th mammary glands and spleen were removed. We isolated lymphocyte from RLN and spleen to analyze by flowcytometry. The number of CD8 T cell showed no change among the groups in the RLN but was significantly lower in the i.duc group in the spleen. The number of CD62LLOW+ T cell in 5FU treated side of RLN was significantly higher compared to the IV and NT group. On the other hand the number of CD62LLOW+ was low in the spleen. To study the population of systemic memory T cell in the peripheral blood, blood was harvested into Trucount tubes and analyzed with memory T cells surface marker. Twenty week parous mice was administered 5FU i.duc injection only to the left side, IV injection, or NT every 4weeks for a total of 3 times. Cancer cells were injected into the left side of 4th mammary gland fat pad of the mice at 16 weeks after initial procedure. One hundred μl blood was collected 5 days before and 5 days after cancer cell injection. The mice treated by i.duc 5FU showed increased numbers of CD95+/CD62L low effector memory T (TEM) cells, whereas the mice treated with IV 5FU did not recruit TEM cells significantly. There was no significant change in CD95+/CD62L high central memory T cells before and after treatment and among the groups. In summary, i.duc administration of 5FU into mammary glands effectively induced immune effector cells and prevented mammary tumor growth in neu/N transgenic mice. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-456. doi:10.1158/1538-7445.AM2011-LB-456

Obesity is associated with inflammation and elevated aromatase expression in the mouse mammary gland.

Elevated circulating estrogen levels are associated with increased risk of breast cancer in obese postmenopausal women. Following menopause, the biosynthesis of estrogens through CYP19 (aromatase)-mediated metabolism of androgen precursors occurs primarily in adipose tissue, and the resulting estrogens are then secreted into the systemic circulation. The potential links between obesity, inflammation, and aromatase expression are unknown. In both dietary and genetic models of obesity, we observed necrotic adipocytes surrounded by macrophages forming crown-like structures (CLS) in the mammary glands and visceral fat. The presence of CLS was associated with activation of NF-κB and increased levels of proinflammatory mediators (TNF-α, IL-1β, Cox-2), which were paralleled by elevated levels of aromatase expression and activity in the mammary gland and visceral fat of obese mice. Analyses of the stromal-vascular and adipocyte fractions of the mammary gland suggested that macrophage-derived proinflammatory mediators induced aromatase and estrogen-dependent gene expression (PR, pS2) in adipocytes. Saturated fatty acids, which have been linked to obesity-related inflammation, stimulated NF-κB activity in macrophages leading to increased levels of TNF-α, IL-1β, and Cox-2, each of which contributed to the induction of aromatase in preadipocytes. The discovery of the obesity → inflammation → aromatase axis in the mammary gland and visceral fat and its association with CLS may provide insight into mechanisms underlying the increased risk of hormone receptor-positive breast cancer in obese postmenopausal women, the reduced efficacy of aromatase inhibitors in the treatment of breast cancer in these women, and their generally worse outcomes. The presence of CLS may be a biomarker of increased breast cancer risk or poor prognosis.

High-protein diet during gestation and lactation affects mammary gland mRNA abundance, milk composition and pre-weaning litter growth in mice

We evaluated the effect of a high-protein diet (HP) on pregnancy, lactational and rearing success in mice. At the time of mating, females were randomly assigned to isoenergetic diets with HP (40% w/w) or control protein levels (C; 20%). After parturition, half of the dams were fed the other diet throughout lactation resulting in four dietary groups: CC (C diet during gestation and lactation), CHP (C diet during gestation and HP diet during lactation), HPC (HP diet during gestation and C diet during lactation) and HPHP (HP diet during gestation and lactation). Maternal and offspring body mass was monitored. Measurements of maternal mammary gland (MG), kidney and abdominal fat pad masses, MG histology and MG mRNA abundance, as well as milk composition were taken at selected time points. HP diet decreased abdominal fat and increased kidney mass of lactating dams. Litter mass at birth was lower in HP than in C dams (14.8 v. 16.8 g). Dams fed an HP diet during lactation showed 5% less food intake (10.4 v. 10.9 g/day) and lower body and MG mass. On day 14 of lactation, the proportion of MG parenchyma was lower in dams fed an HP diet during gestation as compared to dams fed a C diet (64.8% v. 75.8%). Abundance of MG α-lactalbumin, β-casein, whey acidic protein, xanthine oxidoreductase mRNA at mid-lactation was decreased in all groups receiving an HP diet either during gestation and/or lactation. Milk lactose content was lower in dams fed an HP diet during lactation compared to dams fed a C diet (1.6% v. 2.0%). On days 14, 18 and 21 of lactation total litter mass was lower in litters of dams fed an HP diet during lactation, and the pups’ relative kidney mass was greater than in litters suckled by dams receiving a C diet. These findings indicate that excess protein intake in reproducing mice has adverse effects on offspring early in their postnatal growth as a consequence of impaired lactational function.

Abstract CN12-02: The prostaglandin-aromatase connection: Implications for breast carcinogenesis

Abstract More than 40,000 women in the U.S. die each year of metastatic breast cancer. Notably, obese postmenopausal women are at increased risk of developing hormone receptor (HR)-positive breast cancer. Approximately two-thirds of breast cancer patients have tumors that express estrogen receptors and require estrogen for tumor growth. Estrogens are synthesized from androgens in a reaction catalyzed by cytochrome P450 aromatase (aromatase), encoded by the CYP19 gene. After menopause, peripheral aromatization in adipose tissue is largely responsible for estrogen production. Thus, the increased breast cancer risk in obese postmenopausal women is believed to be attributable, at least in part, to elevated levels of circulating estrogen related to both increased adipose tissue and elevated aromatase expression. Given the epidemic of overweight and obese subjects in the U.S., there is a pressing need to develop mechanism-based strategies to reduce the cancer risk among this sector of the population. Numerous studies have been performed to elucidate the mechanisms that regulate the transcription of CYP19. Several groups have suggested that prostaglandin E2 (PGE2) is a clinically important inducer of aromatase. PGE2 stimulates cAMP-dependent signaling leading to enhanced CYP19 transcription. Cyclooxygenases (COX) catalyze the first step in the synthesis of PGE2 from arachidonic acid. Positive correlations have been detected between COX and aromatase expression in human breast cancer specimens. In mice which express a mammary-targeted COX-2 transgene, increased PGE2 and aromatase levels were observed. Silencing of 15-hydroxyprostaglandin dehydrogenase, the key enzyme responsible for inactivating PGE2, upregulated aromatase. Recently, the tumor suppressor, BRCA1, was found to negatively regulate CYP19 expression. This inhibitory effect of BRCA1 on aromatase expression was relieved by PGE2. Finally, two observational studies found that use of aspirin, an inhibitor of PGE2 production, was associated with a reduced risk of hormone receptor-positive breast cancer. Collectively, these findings provide a strong rationale for targeting the PGE2-aromatase axis as a risk reduction strategy. In both dietary and genetic models of obesity, evidence will be presented that obesity causes significant inflammation in the mammary gland and visceral fat. Elevated levels of proinflammatory mediators (TNF-alpha, IL1-Beta and COX-2) were found in these inflamed tissues. Levels of aromatase mRNA and activity were also increased in both the mammary gland and visceral fat of obese mice and paralleled the increased levels of proinflammatory mediators. Complementary in vitro studies suggested that the observed increase in aromatase reflects, in part, a paracrine mechanism involving both activated macrophages and adipocytes. Silencing or inhibiting COX-2 attenuated the induction of aromatase. Collectively, these findings strongly suggest the possibility that obesity-related inflammation in both the mammary gland and visceral fat contributes to elevated aromatase activity and thereby an increased risk of HR-positive breast cancer. Importantly, our findings provide the basis for future studies to determine whether pharmacological and/or dietary strategies can be used to disrupt the obesity-inflammation-aromatase axis to reduce the risk of HR-positive breast cancer in obese postmenopausal women. Citation Information: Cancer Prev Res 2010;3(12 Suppl):CN12-02.

Effect of caffeine on the metabolic responses of lipolysis and activated sweat gland density in human during physical activity

This study measured caffeine-induced changes in activated sweat gland density (ASGD) and fat oxidation using a randomized crossover design with 10 healthy volunteers given caffeine (Caffe-I, 3 mg/kg ingested 30 min before experiment) and non-caffeine (No-Caff). Subjects were 173.0±3.2 cm in height, 72.5±4.3 kg in weight, and 21.5±2.5 years in age. All experiments were performed in an automated climate chamber (24.0±0.5°C, relative humidity 50±3%, air velocity less than 1 m/sec) between 2–5 p.m. The ASGD on the chest, upper arm, upper back, and lower back were measured (after 30 min running at 60% VO2max), and blood samples were taken (at 40 min before, immediately before and after 30 min running). Activated sweat gland density levels were higher in Caffe-I (Chest p<0.05 and U-Back p<0.01) and free fatty acids (FFA) were higher in Caffe-I compared to No-Caff immediately before (p<0.05) and after running (p<0.01). In summary, caffeine increases ASGD and FFA by stimulating the sympathetic nervous system and increasing of lipolysis.

Abstract 2873: Elucidation of dietary omega-6/omega-3 ratio effects on chemopreventive activity using mammographic density as a surrogate marker in rats

Abstract Although beneficial effects of omega-3 fatty acids in breast cancer prevention have been reported, the optimal dietary omega-6/omega-3 (n-6/n-3) ratio for chemoprevention has not been established. Although n-6 and n-3 fatty acids are essential for the normal development and maintenance of biological function, both classes of fatty acid share metabolizing enzymes and thus have the potential to antagonize each other's metabolism. Therefore, experiments were undertaken to elucidate the ratio(s) of dietary n-6/n-3 that is optimal for chemoprevention in the absence of adverse effects on normal development using mammographic density as a surrogate marker for breast cancer risk. Female Sprague Dawley rats (n=9/group, 32 days of age) were randomized to one of 7 dietary formulations that varied in their ratio of n-6/n-3 as follows: 25:1, 10:1, 5:1, 1:1, 1:5, 1:10, 1:25. The percent of dietary calories from fat was 30% across all 7 formulations. Mammary gland density was determined via digital analyses of abdominal-inguinal mammary gland whole mounts stained with alum carmine. Density was calculated using either area or integrated optical density and expressed as the percent of the mammary gland fat pad occupied by mammary epithelium. Mammographic density was unaffected in the high n-6 arm of the study; whereas, it was significantly reduced in the high omega-3 arm (16%, p&amp;lt;0.03). Efforts are ongoing to determine: 1) whether reduced density is accounted for by effects on cell cycle flux or induction of a pro-apoptotic environment, 2) the extent to which these effects are mediated by altered signaling via activated protein kinase B (Akt), and 3) the mechanism(s) by which the n-6/n-3 ratio alters eicosanoid-mediated bioactivity. Supported by grant KG081632 from the Susan G. Komen Foundation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2873.

Salivary Glands: Echo-Planar versus PROPELLER Diffusion-weighted MR Imaging for Assessment of ADCs

To compare the image distortion and the quantification variation in parotid gland apparent diffusion coefficients (ADCs) on periodically rotated overlapping parallel lines with enhanced reconstruction (PROPELLER) versus echo-planar diffusion-weighted (DW) magnetic resonance (MR) images and to investigate the relationship between parotid gland ADC and parotid gland fat content. This prospective study was approved by a local institutional review board. Written informed consent was obtained from all 33 enrolled healthy volunteers (15 men, 18 women; mean age, 36.4 years +/- 11.8 [standard deviation]). All participants underwent 1.5-T non-fat-saturated and fat-saturated PROPELLER DW MR imaging as well as 1.5-T nonaccelerated and twofold-accelerated echo-planar DW MR imaging. Image distortion on the DW images was qualitatively scored, and parotid ADC was quantitatively analyzed. The correlation between parotid ADC and parotid fat content was evaluated by using linear regression analysis. Wilcoxon signed rank and t tests were used for statistical analysis, with Bonferroni correction applied for multiple comparisons. Echo-planar DW images showed distortion, which was completely eliminated on the PROPELLER DW images. The mean parotid ADCs measured with non-fat-saturated PROPELLER (0.670 x 10(-3) mm(2) +/- 0.149), nonaccelerated echo-planar (0.892 x 10(-3) mm(2) +/- 0.128), twofold-accelerated echo-planar (1.088 x 10(-3) mm(2) +/- 0.124), and fat-saturated PROPELLER (1.307 x 10(-3) mm(2) +/- 0.217) DW imaging differed significantly from one another (P < .001 for all comparisons). Parotid ADC had a significant negative correlation with parotid fat content (x) measured at non-fat-saturated PROPELLER DW imaging: ADC = -0.0087x + 1.1173 (r = 0.80, P < .001). PROPELLER DW imaging pulse sequences can yield distortion-free images for parotid ADC measurements and enable quantitative evaluation of the relationship between parotid ADC and parotid fat content. http://radiology.rsna.org/lookup/suppl/doi:10.1148/radiol.2531082228/-/DC1.

Limited thymectomy for stage I or II thymomas

Background Once an anterior mediastinal tumor has been diagnosed as a thymoma, complete excision including the thymic gland and perithymic fat is currently the procedure of choice. However, little is known about the clinical outcome of grossly encapsulated thymomas excised only with the surrounding tissue while leaving a part of the thymic gland. Methods A retrospective historical comparative study was conducted on 79 patients who had received surgery for stage I ( n = 25) or stage II ( n = 54) thymomas. Total thymectomy was performed in 61 patients (Total Thymectomy Group), whereas resection of tumors with only the surrounding tissue was carried out in 18 (Limited Thymectomy Group). The follow-up interval was longer in the Limited Thymectomy Group because these patients were treated longer ago (104.2 ± 58.1 months vs 67.3 ± 54.8 months, p < 0.05). Results One case in the Limited Thymectomy Group showed postoperative myasthenia gravis (5.6%). Two patients with multiple thymomas (2.5%) were treated with total thymectomy. One case in the Limited Thymectomy Group, which had been diagnosed as Masaoka stage II and WHO type B3 at initial surgery, recurred. None died of tumor progression in this study. Disease free survival rates at 10 years did not differ between the Limited Thymectomy and Total Thymectomy Groups (85.7% and 82.0%, respectively). There were no statistical differences in the incidence of postoperative myasthenia gravis and disease free survival between the two groups. Conclusion Resection of thymomas with surrounding tissue instead of total thymectomy can be indicated for stage I or II thymomas in light of disease free and overall survival, post-operative onset of MG, and the incidence of multiple lesions.

Anti‐endoglin monoclonal antibodies are effective for suppressing metastasis and the primary tumors by targeting tumor vasculature

Anti-metastatic activity of an antitumor agent is exceedingly important because metastasis is the primary cause of death for most solid cancer patients. In this report, we show that 3 anti-endoglin (ENG) monoclonal antibodies (mAbs) SN6a, SN6j and SN6k which define individually distinct epitopes of ENG of tumor vasculature are capable of suppressing tumor metastases in the multiple metastasis models. The metastasis models were generated by i.v., s.c. (into flank) or mammary gland fat pad injection of 4T1 murine mammary carcinoma cells and splenic injection of two types of colon26 murine colorectal carcinoma cells. Individual mAbs were injected i.v. via the tail vein of mice. SN6a and SN6j effectively suppressed the formation of metastatic colonies of 4T1 in the lung in all of the three 4T1 metastatic models. In addition, these mAbs were effective for suppressing the primary tumors of 4T1 in the skin and mammary fat pad. These mAbs effectively suppressed microvessel density and angiogenesis in tumors as measured by the Matrigel plug assay in mice. No significant side effects of the administered mAbs were detected. Furthermore, SN6a and SN6j extended survival of the tumor-bearing mice. SN6j, SN6k and their immunoconjugates with deglycosylated ricin A-chain were all effective for suppressing hepatic metastasis of colon26. The findings in the present study are clinically relevant in view of the ongoing clinical trial of a humanized (chimerized) form of SN6j.