Determination of P-glycoprotein, MDR-related protein 1, breast cancer resistance protein, and lung-resistance protein expression in leukemic stem cells of acute myeloid leukemia

Background: The most primitive leukemic precursor in acute myeloid leukemia (AML) is thought to be the leukemic stem cell (LSC), which retains the properties of self-renewal and high proliferative capacity and quiescence of the hematopoietic stem cell. LSC seems to be immunophenotypically distinct and more resistant to chemotherapy than the more committed blasts. Considering that the multidrug resistance (MDR) constitutive expression may be a barrier to therapy in AML, we have investigated whether various MDR transporters were differentially expressed at the protein level by different leukemic subsets. Methods: The relative expression of the drug-efflux pumps P-gp, MRP, LRP, and BCRP was evaluated by mean fluorescence index (MFI) and the Kolmogorov-Smirnov analysis (D values) in five leukemic subpopulations: CD34(+)CD38(-)CD123(+) (LSCs), CD34(+)CD38(+)CD123(-), CD34(+)CD38(+)CD123(+), CD34(+)CD38(+)CD123(-), and CD34(-) mature cells in 26 bone marrow samples of CD34(+) AML cases. Results: The comparison between the two more immature subsets (LSC versus CD34(+)CD38(-)CD123(-) cells) revealed a higher P-gp, MRP, and LRP expression in LSCs. The comparative analysis between LSCs and subsets of intermediate maturation (CD34(+)CD38(+)) demonstrated the higher BCRP expression in the LSCs. In addition, P-gp expression was also significantly higher in the LSC compared to CD34(+)CD38(+)CD123(-) subpopulation. Finally, the comparative analysis between LSC and the most mature subset (CD34(-)) revealed higher MRP and LRP and lower P-gp expression in the LSCs. Conclusions: Considering the cellular heterogeneity of AML, the higher MDR transporters expression at the most immature, self-renewable, and quiescent LSC population reinforces that MDR is one of the mechanisms responsible for treatment failure. (C) 2008 Clinical Cytometry Society.

Cytotoxic effects of butanolic extract from Pfaffia paniculata (Brazilian Ginseng) on cultured human breast cancer cell line MCF-7

Roots of Pfaffia paniculata have been well documented for multifarious therapeutic values and have also been used for cancer therapy in folk medicine. This study has been performed in a human breast tumor cell line, the MCF-7 cells. These are the most commonly used model of estrogen-positive breast cancer, and it has been originally established in 1973 at the Michigan Cancer Foundation from a pleural effusion taken from a woman with metastatic breast cancer. Butanolic extract of the roots of P. paniculata showed cytotoxic effect MCF-7 cell line. as determined with crystal violet assay, cellular death with acridine orange/ethidium bromide staining, and cell proliferation with immunocytochemistry of bromodeoxyuridine (BrdU). Subcellular alterations were evaluated by electron microscopy. Cells treated With butanolic extract showed degeneration of cytoplasmic components and profound morphological and nuclear alterations. The results show that this butanolic extract indeed presents cytotoxic substances, and its fractions merit further investigations. (C) 2008 Elsevier GmbH. All rights reserved.

Sensory acceptability of raw and extruded bovine rumen protein in processed meat products

The use of bovine rumen protein (raw and extruded) as a replacement for extruded soy protein concentrate in three meat products (pork sausage, chicken hamburger, and kibbe) was investigated. Similarity between rumen and soy protein meat products was assessed using triangle tests and sensory acceptability evaluated by consumer panelists using a nine-point hedonic scale. The addition of raw rumen protein was detected in all meat product types tested, while extruded rumen protein was only detected in kibbe. The addition of raw rumen protein decreased the acceptability of pork sausage aroma and flavor, but improved kibbe appearance, texture and overall acceptability. The addition of extruded rumen protein reduced the acceptability of chicken hamburger texture, but improved pork sausage flavor. Replacement of soy protein by bovine rumen protein is feasible based upon sensory results, but depended upon its form and the type of meat product to which it was added. (C) 2011 Elsevier Ltd. All rights reserved.

Dietary practices and nutritional status of 0-24-month-old children from Brazilian Amazonia

Objective: To assess the nutritional status and dietary practices of 0-24-month-old children living in Brazilian Amazonia. Design: Cross-sectional study. Information oil children`s dietary intakes was obtained from diet history data. Weight and length Were measured for anthropometric evaluation. Fe status Was assessed Using fasting venous blood samples; Hb, serum ferritin and soluble tranferrin receptor concentrations were measured. Setting: The towns of Assis Brasil and Acrelandia in the state of Acre, north-west Brazil. Subjects: A total of sixty-nine randomly selected 0-24-month-old children. Results: Of these children, 40.3 % were anaemic, 63.1% were Fe-deficient, 28.1% had Fe-deficiency anaemia and 11.6% were stunted. Breast-feeding was initiated by 97.1% of mother followed by early feeding with complementary foods. The dietary pattern reflected a high intake of carbohydrate-rich foods and cow`s milk, with irregular intakes Of fruit, Vegetables and meat. All infants and 92.3% of toddlers were at risk Of inadequate Fe intakes. Fe from animal foods contributed Oil average 0.5% and 14.3% to total dietary Fe intake among infants and toddlers, respectively. Conclusions: Poor nutritional status and inadequate feeding practices in this study population reinforce the importance of exclusive breast-feeding during the first 6 months of life. Greater emphasis is required to improve the bioavailability of dietary Fe during complementary feeding practices.

The influence of maternal factors on the concentration of vitamin A in mature breast milk

Background & aims: This study evaluated the relationship between vitamin A concentration in maternal milk and the characteristics of the donors of a Brazilian human milk bank. Material and methods: A total of 136 donors were selected in 2003-2004 for micronutrient determinations in breast milk and blood, anthropometric measurements and investigation of obstetric, socioeconomic-demographic factors, and life style. Maternal serum/milk samples were obtained for vitamin A, iron, copper, and zinc determinations. Vitamin A concentrations in breast milk and blood were assessed by high-performance liquid chromatography. Copper, zinc and iron concentrations in breast milk, and copper and zinc concentrations in blood were detected by atomic emission spectrophotometry. Serum ceruloplasmin and serum iron were determined, respectively, by nephelometry and colorimetry. A linear regression model assessed the associations between milk concentrations of vitamin A and maternal factors. Results: Vitamin A in milk presented positive associations with iron in milk (p < 0.001), serum retinol (p = 0.03), maternal work (p = 0.02), maternal age (p = 0.02). and oral contraceptive use (p = 0.01), and a negative association with % body fat (p = 0.01) (R(2) = 0.47). Conclusion: These results suggest that some nutritional, obstetric, and socioeconomic-demographic factors may have an effect on mature breast milk concentrations of vitamin A in apparently healthy Brazilian mothers. (C) 2009 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

Outcome Analysis of Immediate and Delayed Conservative Breast Surgery Reconstruction With Mastopexy and Reduction Mammaplasty Techniques

Background: Bilateral mammaplasty or mastopexy is frequently used for oncoplastic objectives. However, little information has been available regarding outcome following immediate and delayed reconstruction. Method: Patients were divided into Group I (immediate reconstruction) and Group II (delayed reconstruction). Retrospective review was performed to compare complications, length of hospital stay, revision surgeries, and satisfaction. The associations between the complications with potential risk factors (timing, age, body mass index, smoking, and comorbid medical conditions) were analyzed. Results: There were a total of 144 patients with a mean follow-up of 47 months. Of the 106 patients in Group I, complications occurred in 24 (22.6%), skin necrosis was observed in 7.5%, fat necrosis in 5.6%, and 6.6% patients developed local recurrence. Mean period of hospitalization was 1.89 days. Of the 38 patients of the Group II, complications occurred in 12 (31.5%), skin necrosis was observed in 7 (18.4%), fat necrosis in 4 (10.5%), and 5.2% patients developed local recurrence. Mean period of hospitalization was 1.35 days. Increased length of hospital stay greater than 1 day (P < 0.001) and the number of revision surgeries (P = 0.043) were associated with the timing of the reconstruction. In univariate analysis, no difference between groups was found with respect to complication incidence (P = 0.275); however, after adjusting for other risk factors, the probability of complications tend to be higher for Group II (OR = 2.65; 95% confidence interval - 1.01-7.00; P = 0.049). Conclusions: On the basis of the results of our study, the probability of complications tends to be higher for delayed reconstructions, and it is demonstrated that obesity and smoking are risk factors for complications. Ultimately, these data may facilitate the provision of individualized risk information for shared medical decision-making.

Immediate reconstruction following breast-conserving surgery: Management of the positive surgical margins and influence on secondary reconstruction

The efficacy of breast-conserving Surgery for the local control of early breast cancer has been repeatedly evidenced. Although immediate reconstruction following breast-conserving Surgery has been described, little information is available regarding surgical management in reoperative settings due to positive margins. We studied the influence of intraoperatively assessed and postoperatively controlled Surgical margin status on the type of breast-conserving Surgery and report our results regarding complications in a reoperative breast reconstruction scenario. All patients were seen by a multidisciplinary team who recommended breast-conserving surgery. According to the breast volume, ptosis and tumor size/location, the patients were also evaluated by a plastic Surgeon, who recommended reconstruction with the appropriate technique. Intraoperative assessment Of Surgical margins was determined by histological examination of frozen sections. The mean follow-up time was 48 months. Two hundred and eighteen patients (88.5%) underwent breast-conserving Surgery and immediate reconstruction. Twelve (5.5%) patients had a positive tumor margin after review of the permanent section. All patients underwent re-exploration. In 1.3%, a second reconstructive technique was indicated and in 2.2% a skin-sparing mastectomy with total reconstruction was performed. Our findings support the important role of the intraoperative assessment of surgical margins and its interference in the selection of reconstruction techniques and negative margins: however, it will not guarantee complete excision of the tumor. Success depends on coordinated planning with the oncologic Surgeon and careful intraoperative management, (C) 2008 Elsevier Ltd. All rights reserved.

Immediate conservative breast surgery reconstruction with perforator flaps: New challenges in the era of partial mastectomy reconstruction?

Introduction: Although the use of local flaps in conservative breast surgery (CBS) reconstruction is a reliable technique, little information has been available regarding outcome following the use of perforator flaps. The purpose of this study is to analyze the feasibility, surgical planning and outcome following CBS reconstruction with intercostal artery perforator (ICAP) flap. Patients/methods: Thirteen patients underwent CBS reconstructions with an ICAP flap. These flaps were raised from adjacent tissue located on the lateral and thoracic region and based on perforators originating from the costal and muscular segment of the intercostal vessels. The technique was indicated in patients with small/moderate volume breasts. Results: Mean time of follow-up was 32 months. Flap complications were evaluated and information on patient satisfaction were collected. 61.5 percent had tumors located in the lower-outer quadrants and 69.2 percent had tumors measuring 2 cm or less (T1). Complications occurred in 3 patients (23%), including wound dehiscence in 2 patients and fat necrosis in one. All cases were treated by conservative approach with a good result. No flap loss or wound infection were reported. 90 percent were either satisfied or very satisfied with their result. Conclusion: The ICAP flap is a reliable technique for immediate CBS reconstruction. The technique is advantageous because it does not require the use of muscle transfer, with minimum donor site morbidity. Success depends on patient selection, coordinated planning with the oncological surgeon and careful intra-operative management. (C) 2011 Elsevier Ltd. All rights reserved.

Patterns of food acquisition in Brazilian households and associated factors: a population-based survey

Objective: To identify food acquisition patterns in Brazil and relate them to the sociodemographic characteristics of the household. Design: A cross-sectional national Household Budget Survey (HBS). Principal component factor analysis was used to derive food patterns (factors) on the basis of the acquisition of food classified into thirty-two food groups. Setting: The source of data originates from the 2002-2003 HBS carried out by the Brazilian Institute of Geography and Statistics between June 2002 and July 2003 using a representative sample of all Brazilian households. Subject: A total of 48 470 households allocated into 443 strata of households that were geographically and socio-economically homogeneous as a study unit. Results: We identified two patterns of food acquisition. The first, named `dual`, was characterized by dairy, fruit, fruit juice, vegetables, processed meat, soft drinks, sweets, bread and margarine, and by inverse correlations with Brazilian staple foods. In contrast, the second pattern, named `traditional`, was characterized by rice, beans, manioc, flour, milk and sugar. The `dual` pattern was associated with higher household educational level, income and the average age of adults on the strata, whereas the `traditional` presented higher loadings in less-educated households and in the rural setting. Conclusions: Dietary patterns described here suggest that policies and programmes to promote healthy eating need to consider that healthy and non-healthy foods may be integral in the same pattern.

Iron Concentrations in Breast Milk and Selected Maternal Factors of Human Milk Bank Donors

The aim of this study was to evaluate the relationship between iron concentration in mature breast milk and characteristics of 136 donors of a Brazilian milk bank. Iron, vitamin A, zinc, and copper concentrations were assessed in human milk and maternal blood. Data were collected on maternal anthropometrics, obstetric, socioeconomic, demographic, and lifestyle factors. Iron, zinc, and copper in milk and zinc and copper in blood were detected by spectrophotometry. Vitamin A in milk and blood was determined by high-performance liquid chromatography. Hemoglobin was measured by electronic counting and serum iron and ferritin by colorimetry and chemoluminescence, respectively. Transferrin and ceruloplasmin were determined by nephelometry. According to multivariate linear regression analysis, iron in milk was positively associated with vitamin A in milk and with smoking but negatively associated with timing of breast milk donation (P < .001). These results indicate that iron concentration in milk of Brazilian donors may be influenced by nutritional factors and smoking. J Hum Lact. 26(2):175-179

PHYSICAL AND CHEMICAL CHARACTERISTICS OF FROZEN GROUND BEEF AND AGED BEEF MEAT FROM BOS INDICUS STEERS SUPPLEMENTED WITH alpha-TOCOPHEROL ACETATE

The objective of this study was to evaluate the effects of diet supplementation with vitamin E on the physical and chemical characteristics of ground, frozen and stored or aged Quadriceps femoris (QF) and Longissimus dorsi (LD) muscles from Nellore steers fed high concentrate diets. Muscles were obtained from 24 animals that were 30 months old with a mean live weight of 279 kg. Half of the animals received daily doses of 1,000 mg of alpha-tocopherol acetate (VIT E) per head per day that was added to 100 g of corn meal. The other half received 100 g of corn meal without the antioxidant. Twenty-four hours after slaughtering, QF samples from each animal were ground, frozen and stored for up to 6 months. In addition, 4 samples from the LD of each animal were vacuum packed individually and kept for 21 days. All samples were analyzed to determine the pH, color and water-holding-capacity. The VIT E supplementation improved only the water loss characteristics of frozen ground QF and did not have any positive effect on the physical-chemical characteristics of the aged LD.

Systemic Inflammatory Reaction After Silicone Breast Implant

Systemic inflammation after augmentation mammaplasty with modern silicone implants is not currently recognized. In a prospective controlled study, C-reactive protein and other variables were monitored, aiming to test this hypothesis in a young cohort of patients. Females (18-30 years old, BMI = 18.5-30 kg/m(2), N = 52) were consecutively recruited for breast implant (n = 24, Group I) and for abdominal liposuction (n = 28, Group II/Controls). Patients were interviewed at baseline and followed until 6 months after operation. Variables included demographic and clinical information, surgical outcome, inflammatory markers and autoantibodies. Operations were well tolerated, without surgical or infectious complications. Mean prosthesis size was 258 +/- A 21 ml (range = 220-280) and mean aspirate of liposuction was 1972 +/- A 499 ml (range = 1200-3000). Preoperative, 2-month, and 6-month C-reactive protein concentrations for breast implant patients were 1.3 +/- A 1.2, 4.8 +/- A 3.0, and 4.3 +/- A 6.4 mg/l and for liposuction 3.5 +/- A 2.7, 3.5 +/- A 2.1, and 2.2 +/- A 2.2 mg/l, respectively. Change at 2 months was significant (p = 0.001). Autoantibody investigation failed to reveal remarkable aberrations, except for anticardiolipin elevation, which was nearly symmetrical in the two groups. C-reactive protein levels increased after operation and correlated with proinflammatory and procoagulatory indices. A mild increase in anticardiolipin IgM occurred but differences between populations were lacking. Despite excellent cosmetic outcomes and lack of complications, acute phase reaction could signal ongoing immunogenicity of silicone and long-term monitoring is recommended.

Transfer of antibodies across the placenta and in breast milk from mothers on intravenous immunoglobulin

We studied the levels of immunoglobulins in colostrum, milk and sera from two common variable immunodeficiency (CVID) mothers (M1 and M2), and in sera from their newborn infants. During pregnancy they continued intravenous immunoglobulin therapy (IVIG). Antibody levels from maternal and cord blood collected at delivery and colostrum and milk, collected on the 3rd and 7th post-partum days, respectively, were analyzed. Although cord/maternal blood ratios of total immunoglobulins and subclasses, as well as specific antibodies differed between M1 and M2, both showed good placental transfer of anti-protein and anti-polysaccharide antibodies, despite lower cord/maternal blood ratios in M2. Anti-Streptococcus pneumoniae antibody avidity indexes were similar between paired maternal and cord serum. Both mothers` colostrum and milk samples showed only traces of IgA, and IgM and IgG levels in colostrum were within normal range in M1, whereas M2 presented elevated IgG and low IgM levels, when compared with healthy mothers. The study of colostrum and milk activity showed that they strongly inhibited enteropathogenic Escherichia coli adhesion in vitro. CVID patients must be informed about the relevance of regular IVIG administration during pregnancy, not only for their own health but also for their immune immature offspring. Breast-feeding should be encouraged as colostra from these CVID patients strongly inhibited E. coli adhesion to human epithelial cells thus providing immunological protection plus nutritional and psychological benefits for the infant.

The geodiamolide H, derived from Brazilian sponge Geodia corticostylifera, regulates actin cytoskeleton, migration and invasion of breast cancer cells cultured in three-dimensional environment

We are investigating effects of the depsipeptide geodiamolide H, isolated from the Brazilian sponge Geodia corticostylifera, on cancer cell lines grown in 3D environment. As shown previously geodiamolide H disrupts actin cytoskeleton in both sea urchin eggs and breast cancer cell monolayers. We used a normal mammary epithelial cell line MCF 10A that in 3D assay results formation of polarized spheroids. We also used cell lines derived from breast tumors with different degrees of differentiation: MCF7 positive for estrogen receptor and the Hs578T, negative for hormone receptors. Cells were placed on top of Matrigel. Spheroids obtained from these cultures were treated with geodiamolide H. Control and treated samples were analyzed by light and confocal microscopy. Geodiamolide H dramatically affected the poorly differentiated and aggressive Hs578T cell line. The peptide reverted HsS78T malignant phenotype to polarized spheroid-like structures. MCF7 cells treated by geodiamolide H exhibited polarization compared to controls. Geodiamolide H induced striking phenotypic modifications in Hs578T cell line and disruption of actin cytoskeleton. We investigated effects of geodiamolide H on migration and invasion of Hs578T cells. Time-lapse microscopy showed that the peptide inhibited migration of these cells in a dose-dependent manner. Furthermore invasion assays revealed that geodiamolide H induced a 30% decrease on invasive behavior of Hs578T cells. Our results suggest that geodiamolide H inhibits migration and invasion of Hs578T cells probably through modifications in actin cytoskeleton. The fact that normal cell lines were not affected by treatment with geodiamolide H stimulates new studies towards therapeutic use for this peptide.

Refraction corrected transmission ultrasound computed tomography for application in breast imaging

Purpose: We present an iterative framework for CT reconstruction from transmission ultrasound data which accurately and efficiently models the strong refraction effects that occur in our target application: Imaging the female breast. Methods: Our refractive ray tracing framework has its foundation in the fast marching method (FNMM) and it allows an accurate as well as efficient modeling of curved rays. We also describe a novel regularization scheme that yields further significant reconstruction quality improvements. A final contribution is the development of a realistic anthropomorphic digital breast phantom based on the NIH Visible Female data set. Results: Our system is able to resolve very fine details even in the presence of significant noise, and it reconstructs both sound speed and attenuation data. Excellent correspondence with a traditional, but significantly more computationally expensive wave equation solver is achieved. Conclusions: Apart from the accurate modeling of curved rays, decisive factors have also been our regularization scheme and the high-quality interpolation filter we have used. An added benefit of our framework is that it accelerates well on GPUs where we have shown that clinical 3D reconstruction speeds on the order of minutes are possible.