Dietary Intake of Flaxseed Oil since Early Stages of Life Promotes Femur Quality in Male Rats.

BACKGROUND: Flaxseed oil (FO) is an alpha linolenic acid source important for growth and body development. However, there is little literature on the role of FO in critical stages of bone development and formation. OBJECTIVE: This study evaluated the influence of a diet containing FO on rat femurs. METHODS: After birth, mothers and pups were divided into control and flaxseed groups (n = 6 pups each) fed diets containing 7% soybean oil (C) or 7% FO. At 21 days, pups were weaned and separated from the mothers, and control or experimental diets were continued. At 67 days, the following were analyzed: osteocalcin and osteoprotegerin (OPG) levels, bone mineral density (BMD) and content, and bone area; the dimension, BMD, head radiodensity, and biomechanical proprieties of the right femur; and histomorphometric parameters of the left femur. RESULTS: Compared to the C group, the FO group presented (p < 0.05) a lower body mass (-3.7%) and medullary area (-10.1%) and higher osteocalcin (+36.7%), OPG (+52.5%), femur width (+3.8%), absolute mass (+2.3%), femur BMD (+3.6%), head radiodensity (+6.1%), maximum force (+7.4%), breaking strength (+17.3), and cortical thickness (+7.0). CONCLUSION: The FO diet contributed to femur quality in healthy male Wistar rats.

Beverages Rich in Resveratrol and Physical Activity Attenuate Metabolic Changes Induced by High-Fat Diet.

Consumption of saturated fat causes deleterious effects on health, which could be minimized through physical activity and foods with functional characteristics consumption. The aim of the study was to evaluate the beverage rich in resveratrol consumption and physical exercise in gut microbiota, body composition, lipid peroxidation, interleukin-6 (IL6) concentration and systolic blood pressure (SBP) of rats to the high-fat diet. Wistar rats were fed with control diet, high-fat diet (HFD), HFD and 15 mL solution of resveratrol, HFD and 15 mL of grape juice, HFD and 10 mL of red wine. All animals performed the physical training protocol five days a week. Grape juice and red wine composition were analyzed, SBP, body mass, consumption, adiposity and body composition, gut microbiota, lipid peroxidation and inflammation were evaluated. The grape juice (114.8 ± 22.5 mmHG) and red wine (129 ± 15.8 mmHg) groups showed lower SBP when compared to HFD (216.8 ± 20.6 mmHg) (p < 0.0001). The grape juice group (GJG) (39.1 ± 7) had a higher number of microbiota bands DNA when compared to the other groups (p = 0.002). The GJG (33.7 ± 6.7 pg/mL) presented lower concentration IL6 when compared to high-fat group (47.3 ± 16 pg/mL) (p = 0.003). GJG (4.7 ± 1.2 nmol/L) presented a lower concentration of TBARS when compared to control group (6.1 ± 1.4 nmol/L) and resveratrol group (6.6 ± 0.9 nmol/L), and the red wine group (7.4 ± 1.2 nmol/L) had a higher concentration of TBARS when compared to control group and GJG (p = 0.0001). The consumption of these beverages, especially grape juice, together with physical exercise, was able to promote beneficial changes even in the presence of a HFD.

Antioxidant action of alpha lipoic acid on the testis and epididymis of diabetic rats: morphological, sperm and immunohistochemical evaluation.

INTRODUCTION: Chronic hyperglycemia is caused by diabetes mellitus-committed genital morphophysiology, and oxidative stress is one of the main factors involved in this process. Alpha lipoic acid (ALA) can prevent metabolic and morphological changes in diabetic individuals. OBJECTIVES: In present study, we evaluated the effects of regular ALA consumption on the spermatogenesis and histoarchitecture in the male genital system of diabetic rats. MATERIALS AND METHODS: Thirty-two Wistar rats were divided into groups: Control (CG); Diabetic Control (DCG), receiving commercial diet: ALA Group (ALAG) and Diabetic ALA Group (DALAG), fed diets with added ALA (300 mg/Kg bw). The diabetic groups received a single injection of streptozotocin (60 mg/kg). After sixty days of the diet, the animals were euthanized, and semen, testis and epididymis samples were collected. A histomorphometric analysis was performed to determine the epithelial height, tubular and luminal diameter, tubular and luminal area of seminiferous tubules and each epididymal region. Sertoli cells were evidenced using the antivimentin antibody and were quantified. The results were statistically analyzed by the ANOVA test. RESULTS: At the end of the experiment, the DALAG glycemia was signifi cantly lower than DCG. The histomorphometric parameters of the seminiferous and epididymal tubules did not show improvement in the DALAG. However, there was an improvement in the DALAG in terms of the concentration, motility and percentage of spermatic pathologies, as well as in the number of Sertoli cells (p<0.001). CONCLUSIONS: The results demonstrated that supplementation with the ALA antioxidant retards testicular lesions and preserve the process of spermatogenesis in diabetes.

Flaxseed (linum usitatissimum) flour contributes to bone health in adult male rats.

OBJECTIVE: This study evaluated effects of flaxseed flour diet on bone health in adult Wistar male rats at 180 d. METHODS: After birth, pups were randomly assigned: Control (C, n = 12) and flaxseed flour (FF, n = 12) diet during lactation period. At 21 d, pups were weaned and they continued to receive the control and experimental diet, respectively. At 180 d, body length, total bone structure, and femur and lumbar vertebrae were evaluated by dual-energy X-ray absorptiometry. Femoral and lumbar vertebra dimensions, biomechanical properties of femur, and serum osteocalcin were measured. RESULTS: FF produced greater (P <0.05) total (+3.4%) and spine (+5.9%) bone mineral density, total (+10%) and spine (+12%) bone mineral content, total bone area (+8.7%), and osteocalcin (+82%). As to femur, FF produced greater (P <0.05) width of diaphysis (+8.1%), bone mineral density (+5.2%), maximum force (+10.6%), breaking strength (+12.9%), and stiffness (+23%). CONCLUSION: This experimental study found that a diet containing flaxseed flour contributed to bone mineral density and femur resistance at 180 d.

Effects of flour or flaxseed oil upon testis mass in rats subjected to early weaning.

Study evaluates testis mass in rats subjected to early weaning and subsequently nourished with diet containing flour or flaxseed oil. Pups were weaned for separation from mothers at 14 days (early weaning, EW) and 21 days (control, C). After 21 days, the control group (C60) was nourished with control diet. EW was divided as: control (EWC60), flaxseed flour (EWFF60) and flaxseed oil (EWFO60) group diets for the next 60 days. At 21st and 60th day, body mass, serum cholesterol and triglycerides and testis mass were evaluated. At 21 day, EW group showed lower (p < 0.05) body mass, serum cholesterol and testis mass. At 60 days, EWC60 and EWFO60 groups showed lower (p < 0.05) body mass (vs. C60 and EWFF60). EWFF60 group showed lower (p < 0.05) serum cholesterol (vs. EWC60 and EWFO60) and higher (p < 0.05) testis mass (vs. C60, EWC60 and EWFO60). Flaxseed flour (vs. oil) was associated with higher testis mass following early weaning.

Incorporation of Flaxseed Flour as a Dietary Source for ALA Increases Bone Density and Strength in Post-Partum Female Rats.

In women, bone mass undergoes changes during pregnancy and the postpartum period, which has a risk for subsequent development of osteoporosis. Thus, the present study aims to evaluate the effects of flaxseed flour in femur quality during post-weaning of dam rats. After weaning, the rats were divided into control (C, n = 7) and experimental (F, n = 7) groups treated with a diet containing 25 g of flaxseed flour in the lactating period and 15 g in the maintenance period. After 51 days post-partum, serum hormone, fatty acids composition, bone compartments, computed tomography, and biomechanical analyses were determined. Food intake, length, body mass, hormone analysis, and total bone compartments showed similar results. For biomechanical and computed tomography analysis and fatty acids composition, the F group showed higher maximum force (+12%, p < 0.05), breaking strength (+25%, p < 0.05), rigidity (+17%, p < 0.0001), and femoral head radiodensity (+15%, p < 0.05) and presented lower total polyunsaturated fatty acids (-17%, p < 0.0001) and arachidonic acid (-44%, p < 0.0001) and higher ALA (+695%, p < 0.0001) and EPA (+160%, p < 0.05). Fatty acids composition of flaxseed flour, as well as its protein profile and calcium content, were able to improve the bone quality, which may be associated with lower serum levels of arachidonic acid and higher EPA, showing an anti-inflammatory profile and increased deposition of organic matrix during the post-weaning period, and may result in prevention of future osteoporosis.

Flaxseed reduces epithelial proliferation but does not affect basal cells in induced benign prostatic hyperplasia in rats.

PURPOSE: This study aimed to quantitatively and qualitatively evaluate the effects of a flaxseed-based diet on the histoarchitecture of the prostate of normal Wistar rats and of rats with induced BPH. METHODS: The study included four experimental groups of ten animals each: casein control group (CCG), who were fed a casein-based diet; flaxseed control group (FCG), who were fed a flaxseed-based diet; hyperplasia-induced casein group (HICG), who were fed a casein-based diet; and hyperplasia-induced flaxseed group (HIFG), who were fed a flaxseed-based diet. Hyperplasia was induced by the subcutaneous implantation of silicone pellets containing testosterone propionate. After 20 weeks, the rats were euthanized and their prostate fixed in buffered formalin. Tissue sections were stained with HE, picrosirius red and immunostained for nuclear antigen p63. Histomorphometric analysis evaluated the epithelial thickness, epithelial area, individual luminal area, and total area of prostatic alveoli. RESULTS: The mean epithelial thickness obtained for HIFG and HICG was 16.52 ± 1.65 and 20.58 ± 2.86 µm, respectively. The mean epithelial thickness in HICG was greater than that in the other groups tested. HIFG had a smaller epithelial thickness and lower percentage of papillary projections in the prostatic alveoli. No significant difference was observed between CCG and FCG. The total area and mean alveolar area showed no significant differences between the groups. The number of cells immunostained for p63 was not significantly different between the groups evaluated. CONCLUSION: These results suggest that flaxseed has a protective effect on the prostate epithelium in BPH-induced animals.

Maternal Flaxseed Oil During Lactation Enhances Bone Development in Male Rat Pups.

Flaxseed oil is an alpha linolenic acid source important in the growth and body development stage; furthermore, this acid acts on adipose tissue and bone health. The aim of this study was to evaluate body composition, fatty acid composition, hormone profile, retroperitoneal adipocyte area and femur structure of pups at weaning, whose mothers were fed a diet containing flaxseed oil during lactation. After birth, pups were randomly assigned: control (C, n = 12) and flaxseed oil (FO, n = 12), rats whose mothers were treated with diet containing soybean or flaxseed oil. At 21 days, the pups were weaned and body mass, length, body composition, biochemical parameter, leptin, osteoprotegerin, osteocalcin, fatty acids composition, intra-abdominal fat mass and femur structure were analyzed. FO showed (p < 0.05): higher body mass (+12 %) and length (+9 %); body fat mass (g, +45 %); bone mineral density (+8 %), bone mineral content (+55 %) and bone area (+35 %), osteocalcin (+173 %) and osteoprotegerin (+183 %). Arachidonic acid was lower (p < 0.0001), alpha-linolenic and eicosapentaenoic were higher (p < 0.0001). Intra-abdominal fat mass was higher (+25 %), however, the retroperitoneal adipocytes area was lower (-44 %). Femur mass (+10 %), distance between epiphyses (+4 %) and bone mineral density (+13 %) were higher. The study demonstrates that adequate flaxseed oil content during a lactation diet plays an important role in the development of pups.

Flaxseed flour, compared to flaxseed oil, contributes to femoral structure in male rats subjected to early weaning.

The aim of this study was to analyze whether flaxseed flour or flaxseed oil treatment contributes to femoral structure in male rats subjected to early weaning. Pups were weaned for separation from mothers at 14 days (early weaning, EW) or 21 days (control, C). After 21 days, the control (C60) was fed with the control diet. The EW group was divided based on control (EWC60), flaxseed flour (EWFF60) and flaxseed oil (EWFO60) diets until 60 days. Femoral dimension, bone mineral density (BMD), bone mineral content (BMC), area and biomechanical properties were determined. EWFO60 showed lower (P < 0.05) femur mass. EWC60 and EWFO60 showed lower (P < 0.05) distance between epiphyses, diaphysis width and BMD. BMC was lower (P < 0.05) in EWC60 (vs. C60 and EWFF60). EWC60 and EWFO60 showed lower (P < 0.05) maximum force (vs. C60). Breaking strength was lower (P < 0.05) in EWFO60 (vs. C60). EWFF60 showed higher (P < 0.05) rigidity. Flaxseed flour abbreviated the femoral fragility secondary to early weaning.

Effects of diet containing flaxseed flour (Linum usitatissimum) on body adiposity and bone health in young male rats.

Flaxseed flour has been described as an excellent alpha-linolenic acid source. This study was designed to evaluate the effects of flaxseed flour on body adiposity and bone health in rats fed a flaxseed flour diet during lactation until 90 days. At birth, male Wistar rats were randomly assigned to control (C) and experimental (FF) groups, whose dams were treated with a control or flaxseed flour diet, respectively, during lactation. At 21 days, pups were weaned and fed a control and experimental diet until 90 days. Food intake, body mass and length were evaluated during a 21-90 day period. At 90 days, composition by dual-energy X-ray absorptiometry, serum hormonal profile, intra-abdominal fat mass, and lumbar vertebra and femur analyses was determined. Differences were deemed significant at p < 0.05. The FF group displayed the following (P < 0.05): a higher total lean mass (+7%), a lower total (-16%) and intra-abdominal (-24%) fat mass, a smaller adipocyte area (-30%), a higher femoral mass (+5%), bone mineral density (+5%) and radiodensity (+20%), and a higher maximum force (+10%) and breaking strength (+11%). The flaxseed flour diet displayed functional properties related to body growth maintenance associated with a lower risk of developing metabolic alterations, obesity and bone fragility.

Fat tissue morphology of long-term sex steroid deficiency and estrogen treatment in female rats.

After long-term estradiol deficiency, female rats displayed body mass gain accompanied by an increase in the size of adipocytes, an increase in hyperglycemia, and a decrease in insulinemia. The effects were reversed by daily estradiol treatment. Adiposity was suggested by the increased vascular endothelial growth factor expression in castrated rats, whereas the proliferative effect of estradiol was suggested by the increased fibronectin expression in treated rats.

Evaluation of body development, fat mass and lipid profile in rats fed with high-PUFA and -MUFA diets, after neonatal malnutrition.

Neonatal malnutrition is associated with several features of the metabolic syndrome, later in life. Although the recovery of malnutrition was studied with different high-fat diets, few studies compare the effects of enriched vegetable oil diets, containing PUFA and MUFA, after weaning. Our aim was to evaluate the recovery with soya oil- or rapeseed oil-enriched diet, after malnutrition in rats whose mothers were food restricted (FR) during lactation. Dams were 50 % FR and compared to standard diet-fed dams (control, C). At 21 d, FR offspring had a lower body mass and length. After weaning C and FR offspring were fed a diet containing 7 % soya oil (7 %sC and 7 %sFR), or supplemented with 19 % soya oil (19 %sC or 19 %sFR) or 19 % rapeseed oil (19 %cC or 19 %cFR). The normal animals fed enriched vegetable oil diets had more visceral fat mass, but lower serum TAG and higher HDL-cholesterol. The 19 %FR groups showed significantly less food intake and body development compared to the 7 %sFR, and the same pattern was observed when this group was compared to the C groups. Absolute and relative mass of vital organs and body were lower in the FR groups. Visceral fat depot was lower in 19 %FR than 7 %FR and C groups. Serum glucose, albumin, TAG, cholesterol, leptin and triiodothyronine did not show significant changes. However, 19 %FR groups showed higher HDL-cholesterol and the 19 %sFR group showed lower serum thyroxine. The data suggest that a higher vegetable oil diet in the recovery of neonatal malnutrition ameliorates some features of the metabolic syndrome later in life.

Immuno-biochemical evaluations of phenol and thimerosal as antigen preservatives in Montenegro skin test.

Montenegro skin test (MST) represents the main complementary diagnostic test for tegumentary leishmaniases (TL) in endemic regions. Most antigen formulations used for the MST contain thimerosal as preservative. The Food and Drug Administration (FDA), however, recommended reducing or eliminating thimerosal from vaccines and other biological reagents and the Agência Nacional de Vigilância Sanitária (ANVISA) in Brazil, prohibited the use of mercurial compounds in immunobiologicals. In the search for an alternative stabilizer, phenol and thimerosal were tested as antigen preservatives in MST. Formulations were tested when fresh and after a 12-month storage at 4 degrees C in TL confirmed mice and human patients, and were evaluated for protein constitution by SDS-PAGE, Western blot and anti-gp63 ELISA. In mice, a decrease in the diagnostic effectiveness in merthiolate formulation was observed after a 12-month storage. SDS-PAGE, Western blot and anti-gp63 ELISA analyses showed a degradation of antigen proteins in both formulations after 12-month storage and that phenol-preserved antigen was quantitatively and qualitatively better than the merthiolate-preserved one. In patients, the average of induration diameter was larger in fresh antigens (p<0.05). However, storage time did not jeopardize their diagnostic capacity. No non-specific reactions produced by phenol or merthiolate were observed neither in humans nor in mice. Phenol could be a good alternative to replace the merthiolate in MST, and despite the proteolytic activity, antigens remain viable for at least 12 months.

Immunochemotherapy for cutaneous leishmaniasis: a controlled trial using killed Leishmania (Leishmania) amazonensis vaccine plus antimonial.

BACKGROUND: Leishmaniasis is endemic in 88 countries in the world, and 350 million individuals are at risk of acquiring the disease. Treatment for American cutaneous leishmaniasis (ACL) is long, expensive, and associated with important side-effects. METHODS: In this double-blind, placebo-controlled study, we treated 102 patients with ACL using either a combination of a single-strain Leishmania amazonensis killed promastigote vaccine plus a half dose of meglumine antimoniate, or placebo plus the same half dose regimen of meglumine antimoniate, in 10-day series followed by 10-day intervals. RESULTS: Of the 47 patients in the experimental arm, 47 (100%) were cured after four series of treatment, compared to four of 49 (8.2%) in the control group (P < 0.0001). Six patients were lost to follow-up. CONCLUSIONS: The combination of a single-strain Leishmania (Leishmania) amazonensis killed promastigote vaccine with a half dose regimen of antimonial is highly effective for the treatment of ACL.

Vaccination of C57BL/10 mice against cutaneous Leishmaniasis using killed promastigotes of different strains and species of Leishmania.

Antigenic extracts from five Leishmania stocks were used to vaccinate C57BL/10 mice. The Leishvacin(R) and PH8 monovalent vaccine yielded the highest IFN-gamma levels in the supernatants of spleen cell culture from vaccinated animals. Each single strain immunized group showed evidence of protective immunity six months after the challenge with promastigotes of Leishmania (Leishmania) amazonensis. No differences were detected between the vaccinated groups. It can be concluded that vaccines composed of single Leishmania stocks can provide protection to C57BL/10 mice against L. (L.) amazonensis infection.