Study protocol to investigate biomolecular muscle profile as predictors of long-term urinary incontinence in women with gestational diabetes mellitus.

BACKGROUND: Pelvic floor muscles (PFM) and rectus abdominis muscles (RAM) of pregnant diabetic rats exhibit atrophy, co-localization of fast and slow fibers and an increased collagen type I/III ratio. However, the role of similar PFM or RAM hyperglycemic-related myopathy in women with gestational diabetes mellitus (GDM) remains poorly investigated. This study aims to assess the frequency of pelvic floor muscle disorders and pregnancy-specific urinary incontinence (PS-UI) 12 months after the Cesarean (C) section in women with GDM. Specifically, differences in PFM/RAM hyperglycemic myopathy will be evaluated. METHODS: The Diamater is an ongoing cohort study of four groups of 59 pregnant women each from the Perinatal Diabetes Research Centre (PDRC), Botucatu Medical School (FMB)-UNESP (São Paulo State University), Brazil. Diagnosis of GDM and PS-UI will be made at 24-26 weeks, with a follow-up at 34-38 weeks of gestation. Inclusion in the study will occur at the time of C-section, and patients will be followed at 24-48 h, 6 weeks and 6 and 12 months postpartum. Study groups will be classified as (1) GDM plus PS-UI; (2) GDM without PS-UI; (3) Non-GDM plus PS-UI; and (4) Non-GDM without PS-UI. We will analyze relationships between GDM, PS-UI and hyperglycemic myopathy at 12 months after C-section. The mediator variables to be evaluated include digital palpation, vaginal squeeze pressure, 3D pelvic floor ultrasound, and 3D RAM ultrasound. RAM samples obtained during C-section will be analyzed for ex-vivo contractility, morphological, molecular and OMICS profiles to further characterize the hyperglycemic myopathy. Additional variables to be evaluated include maternal age, socioeconomic status, educational level, ethnicity, body mass index, weight gain during pregnancy, quality of glycemic control and insulin therapy. DISCUSSION: To our knowledge, this will be the first study to provide data on the prevalence of PS-UI and RAM and PFM physical and biomolecular muscle profiles after C-section in mothers with GDM. The longitudinal design allows for the assessment of cause-effect relationships between GDM, PS-UI, and PFMs and RAMs myopathy. The findings may reveal previously undetermined consequences of GDM.

Intake, digestibility, ingestive behavior, and nitrogen balance of goats fed with diets containing residue from tamarind fruit.

This study is aimed at determining the maximum inclusion level of tamarind (Tamarindus indica) residues in the diet of goats on intake, digestibility, ingestive behavior, and nitrogen (N) balance. Twenty-four crossbred (Boer × undefined breed) castrated goat kids (5 months old and with an initial weight of 23.9 ± 0.3 kg) were assigned in a completely randomized design (4 treatments and 6 replicates). Diets consisted of Tifton 85 (Cynodon sp.) hay as the roughage (400 g/kg) source and concentrate (600 g/kg); the levels of tamarind residue inclusion were 0.0, 7.0, 14.0, and 21.0% on a dry matter (DM) basis. The experimental period lasted 23 days (15 of adaptation and 8 of sampling). Inclusion of tamarind residue in the goat kid diets did not affect (P > 0.05) the intake and digestibility of DM, crude protein, neutral detergent fiber (NDF), and total digestible nutrient; intake of N, urinary N, and retained N (g/day); time spent ruminating; numbers of times/day feeding, ruminating, or idling; eating efficiency of DM and NDF; number of boluses/day; and amount (g) of DM/bolus. However, there were a linear reduction in ether extract digestibility (P = 0.011) and a linear decreasing trend in non-fibrous carbohydrate digestibility (P = 0.083). The addition of tamarind residue had a positive linear effect (P = 0.041) on the time spent feeding and promoted a decreasing linear trend for the time spent idling (P = 0.063). It is recommended to include the residue from tamarind fruit at a level of 21% in diets for goat kids, as it does not affect nutrient intake and digestibility and the N balance.

Oral fish oil positively influences nutritional-inflammatory risk in patients with haematological malignancies during chemotherapy with an impact on long-term survival: a randomised clinical trial.

BACKGROUND: Studies suggest that the ingestion of fish oil (FO), a source of the omega-3 polyunsaturated fatty acids docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), can reduce the deleterious side-effects of chemotherapy. The aim of this randomised clinical trial was to evaluate the effect of supplementation with oral FO for 9 weeks on nutritional parameters and inflammatory nutritional risk in patients with haematological malignancies during the beginning of chemotherapy. METHODS: Twenty-two patients with leukaemia or lymphoma were randomised to the unsupplemented group (UG) (n = 13) or supplemented group (SG) (n = 9). SG received 2 g/day of fish oil for 9 weeks. Nutritional status, serum acute-phase proteins and plasma fatty acids were evaluated before (T0) and after (T1) the intervention period. Data were analysed using two models; model 1, comprising data from all patients included in the study, and model 2, comprising data from UG patients with no increase in the proportions of EPA and DHA in plasma and data from SG patients showing an at least 100% increase in plasma EPA and DHA. RESULTS: SG showed an increased plasma proportion of EPA and DHA in both models. In model 2, C-reactive protein (CRP) and CRP/albumin ratio showed larger reductions in the SG. Overall long-term survival in both models (465 days after the start of the chemotherapy) was higher in the group ingesting fish oil (P < 0.05). CONCLUSIONS: These findings indicate an improved nutritional-inflammatory risk and potential effects on long-term survival in patients with haematological malignancies supplemented with FO during the beginning of chemotherapy.

Analysis of p53 gene polymorphism (codon 72) in symptomatic patients with atherosclerosis.

Atherosclerosis is a multifactorial pathological disease that alters the morphology and function of arterial walls. The atheroma growth leads to vessel hardening and lumen narrowing, limiting the blood flow. The atheroma plaque can eventually break, expose highly thrombogenic material and lead to platelet activation and subsequent formation of a thrombus that may block blood flow in loco, or even leading to obstruction of other vessels with a smaller diameter. This process is one of the main determinants of the clinical manifestations of atherosclerosis, such as coronary artery disease, ischemic stroke, and peripheral arterial disease. Although the inflammatory theory about atherosclerosis is the most renowned one, observations point to common biological characteristics between cancer and atherosclerosis suggesting a possible association between p53 and atherosclerotic diseases. We collected peripheral blood samples from 200 individuals with clinical manifestations of atherosclerotic disease and 100 individuals without manisfestation of the disease to form the control group. DNA was subjected to molecular analysis (PCR) to identify the polymorphism of the p53 gene. We have not found any relationship between the polymorphism of the p53 gene and atherosclerosis in the population studied (P = 0.36). There was no relationship between atherosclerosis, polymorphism of p53 and the variables accounted: smoking habit (P = 0.72, 0.51 and 0.62 for smokers, non-smokers and former smokers respectively), alcohol consumption (P = 0.17 for individuals with drinking habits and 0.38 for those who do not consume alcohol beverage), systemic arterial hypertension (P = 0.60), diabetes mellitus (P = 0.34), and dyslipidemia (P = 0.89). Our population has a high rate of miscegenation and heterozygotes, and according to studies the arginine variant is more related to plaque formation because it induces apoptosis more frequently when compared to the proline variant. According to our results, there is no association between the polymorphism of the p53 gene, atherosclerosis and its risk factors in the population studied.

GSTM1 polymorphism in patients with clinical manifestations of atherosclerosis.

Atherosclerosis is characterized by lesions, called atheroma or atheromatous plaques, in the inner layer of blood vessels, which block the vascular lumen and weaken the underlying tunica media. Several modifiable and non-modifiable risk factors for the development of atherosclerosis exist. The modifiable risk factors include hypertension, smoking, obesity, high LDL and low HDL cholesterol levels, sedentary lifestyle, and stress; the non-modifiable factors include diabetes mellitus, family history of hypertension and heart disease, thrombophilia, sex, age, and genetic factors. The association of polymorphisms in GST with coronary artery disease has been studied since the polymorphisms can affect enzyme activity and contribute to the onset of atherosclerosis. We analyzed polymorphisms in GSTM1 in individuals diagnosed with atherosclerosis as well as in healthy individuals (control group). The frequency of the GSTM1 present genotype in the atherosclerosis group was 1.2 times higher than that observed in the control group. We found no sex- or alcohol-consumption-dependent differences between the occurrences of the present and null genotypes. However, the GSTM1 present genotype occurred in 52.6% individuals with atherosclerosis who reported smoking 20 or more cigarettes per day and in 60% individuals who smoked 10 to 20 cigarettes per day (P = 0.0035). In addition, the GSTM1 present genotype was more frequent in individuals who reported being former smokers - 45.5% in individuals with atherosclerosis who smoked for more than 20 years and 50% each for individuals in the control group who smoked for less than 10 years or for 10 to 20 years, respectively (P = 0.0240).

Atherosclerosis: analysis of the eNOS (T786C) gene polymorphism.

The coronary arteriosclerotic disease is the most common cardiovascular disease. Atherosclerosis affects large- and medium-sized arteries leading to severe thrombosis or artery stenosis that could evolve to myocardial infarction, ischemic stroke, ischemic injury of kidneys and intestines, and several other life-threatening clinical manifestations. Nitric oxide has been shown to be a promising therapeutic agent against cardiovascular diseases. The eNOS gene assumes several important functions, including regulation of vascular tone and regional blood flow, the suppression of vascular smooth muscle cell proliferation, and modulation of leukocyte-endothelium interactions. The T786C polymorphism is an important point mutation, where thymine is changed to cytosine. T786C significantly reduces the activity of the eNOS promoter gene. Two hundred and ninety-seven peripheral blood samples were collected from patients with the previous diagnosis of atherosclerotic disease based on clinical examination and confirmed by imaging methods. Results were compared using the chi-square test and the G-test. In the present study, the TC genotype was more frequent in both case and control groups with no statistically significant difference. Comparing the relation TC/TT and CC genotypes in the case and control groups, there was no statistically significant difference. No significant difference was found when genotypes were analyzed regarding gender and smoking. Our results suggest a strong tendency of the T allele, in single or double dose, to be associated with atherosclerosis that was not confirmed by the scientific data.

Polymorphism of the gene eNOS G894T (Glu298Asp) in symptomatic patients with aterosclerosis.

Atherosclerotic and its cardiovascular complications are responsible for 17.5 million deaths a year, according to the World Health Organization. There is consensus that atherosclerosis involves multiple pathogenic processes initiated by endothelial dysfunction, with inflammation and vascular proliferation determining alterations in the matrix, with consequent formation of the atheromatous plaque and its clinical implications. Risk factors such as hypertension, diabetes mellitus, dyslipidemia, and smoking are widely known. Currently, genotyping, which is not directly related to these factors, is not accepted to estimate the risk of cardiovascular diseases, but strong evidence indicates several polymorphic genes as factors of risk and progression leading to complications of the disease. Among the genes involved, eNOS (endothelial nitric oxide synthase gene), which is responsible for the production of endothelial nitric oxide (an important arterial vasodilator), when presented in polymorphic variation can determine production, malfunction, and predisposition to atherosclerosis. In the present study, we analyzed the G894T polymorphism of the eNOS gene in groups of individuals diagnosed with atherosclerosis and in a control group. We collected 200 blood samples from patients previously diagnosed with atherosclerosis and 100 samples formed the control group. The genotyping analysis for polymorphism of the eNOS gene was determined by PCR. We considered variables such as gender, smoking, smoking history, and alcohol consumption; statistical differences were found in the distribution of case and control groups (P = 0.0378) and in non-smoking patients (P = 0.0263). In the other associations, no statistically significant difference was found. In the population studied, the frequency of the heterozygous genotype (GT) was much higher than in the other populations (GG and TT) in both groups (case and control). The GG genotype showed greater susceptibility to atherosclerosis. Association of the GG genotype in non-smokers also showed greater susceptibility. Gender, alcohol consumption, smoking, and smoking history did not influence atherosclerosis.

Molecular analysis of the GSTT1 gene polymorphism in patients with clinical manifestation of atherosclerosis.

Atherosclerosis is a chronic inflammatory disease formed by the accumulation of lipids in the innermost layer and large-caliber artery (tunica intima). This accumulation, along with platelet factors, stimulates the proliferation of muscle cells in this region. Over than 400 genes may be related to the pathology since they regulate endothelial function, coagulation, inflammation, metabolism of amino acids, lipids, and carbohydrates. Glutathione S-transferases (GST) are enzymes that catalyze the polymorphic detoxification of metabolites produced by oxidative stress within the cells, which is induced by reactive oxygen species. GSTs are one of the defense mechanisms against oxidative stress damage. Due to genetic, cultural, and environmental factors, the rate of atherosclerosis is higher; however, an early diagnosis is crucial for the prevention and treatment of several complications related to the disease. The present study aimed to analyze the frequency of GSTT1 genotypes regarding the presence or absence of the polymorphism in patients with clinical manifestation of atherosclerosis. We collected 200 samples of peripheral blood of patients with the previous diagnosis of atherosclerosis based on clinical examination and imaging, and 100 samples of peripheral blood to compose the control group of patients without clinical manifestation of atherosclerosis. The polymorphism was assessed by PCR and analyzed on the agarose gel stained with 2.0% ethidium bromide. The frequency of the GSTT1 gene polymorphism was compared using the chi-square test (P < 0.05) and the G-test. In the case group, we detected 85.5% of patients with the GSTT1 genotype present and 14.5% of patients with the null genotype. A significant difference was observed between groups (case vs control) for the presence of the GSTT1 polymorphism. According to the analysis of the variable alcohol consumption, we found that in the case group the presence of the GSTT1 gene was higher in individuals who reported not drinking alcohol. In this study, the presence of the GSTT1 gene polymorphism in male patients with atherosclerosis was 1.5 times higher when compared to female patients. Regarding the variable time of smoking, we found that this genotype was more frequent in smokers for both case and control groups.

Association of CYP1A1 (cytochrome P450) MspI polymorphism in women with endometriosis.

Endometriosis is a disease that affects 10 to 15% of the women of reproductive age. It is characterized by the presence of endometrial-like tissues outside of the uterus. Some definitions claim that the functional ectopic tissue is sensitive to the action of hormones. Severity of endometriosis is defined according to a system proposed by the American Society for Reproductive Medicine, which is based on laparoscopic findings. A large number of genetic polymorphisms has been reported for CYP1A1, the gene that is responsible for enzymes involved in stage I detoxification of xenobiotics; this gene is located at 15q22-24, and encodes an isoenzyme that catalyzes the oxidation of polycyclic aromatic hydrocarbons present in phenolic compounds and epoxides. The aim of this study was to analyze the frequency of the MspI polymorphism and its relation to endometriosis. We obtained peripheral blood samples from 52 women with endometriosis (confirmed by laparoscopy) as well as 42 women without endometriosis (control group). In the case group, the women were between 25 and 35 years of age; the age range was between 25 and 57 years old in the control group. Molecular analysis was performed by polymerase chain reaction. We found a significant association (P = 0.039) between the polymorphic allele m1 and endometriosis (32.70%). In conclusion, this study showed that the m1 polymorphism is associated with endometriosis, and that W1/m1 and m1/m1 polymorphisms are more frequently observed in patients with infertility and severe endometriosis.

A 7-month multi-disciplinary healthy lifestyle intervention effectively improved cardiometabolic risk profile of firefighters.

OBJECTIVE: We investigated the effect of a 7-month healthy lifestyle intervention on cardiometabolic risk factors (CMRF) among male career military firefighters (FFs). METHODS: 49 FFs participated in a 7-month workplace multi-disciplinary healthy lifestyle intervention designed to reduce CMRF through exercise, diet, and improved sleep. Medical assessments, accelerometry, and surveys at the beginning and end determined program effectiveness. RESULTS: At the end of the intervention period, there was a significant improvement in measures of body composition and blood glucose. The prevalence of hypertension also decreased significantly (p < 0.01). The 57% of participants who fully adhered to the program had significantly greater improvements across multiple CMRF. Participants increased their physical activity and improved their diet following the intervention. CONCLUSION: This healthy lifestyle intervention was effective in changing behavior and lowering cardiometabolic risk among FFs.

GSTM1 and GSTT1 polymorphisms in endometriosis in women from Goiás, Brazil.

Endometriosis is a gynecologic pathology with a high prevalence and unknown etiology. Therefore, an increasing number of studies has been undertaken to search for associations between endometriosis and alterations or polymorphisms in candidate genes, including glutathione S-transferase mu 1 (GSTM1) and glutathione S-transferase theta 1 (GSTT1). We analyzed the frequency of present/absent polymorphisms of GSTM1 and GSTT1 in 50 women diagnosed with endometriosis and in a control group of 46 women without complaints related to this pathology. The association of these polymorphisms with p53 gene codon 72 was also evaluated within each group, and a higher frequency of absence of GSTM1 (61%) and GSTT1 (45%) genes in the group of women studied, women with endometriosis and control group was found. The contributions of GSTM1 and GSTT1 polymorphisms to the proliferation of endometriosis were not statistically significant, but the analysis of pathology and the association of GSTM1 and GSTT1 gene polymorphisms with p53 codon 72 revealed statistical significance.

GSTM1 polymorphism in patients with primary open-angle glaucoma.

Primary open-angle glaucoma (POAG) is characterized by loss of retinal ganglion cells, optic nerve damage and irreversible loss of visual field. Glaucoma is the second leading cause of blindness worldwide. It was estimated that in 2010 there were about 60.5 million glaucoma cases worldwide; among these patients, 4.5 million will become bilaterally blind. Glutathione S-transferases (GST) are a group of drug-metabolizing enzymes of phase-II that act in the detoxification of xenobiotics and inactivate end-products formed as secondary metabolites during oxidative stress. Through PCR amplification, we analyzed the GSTM1 gene in DNA samples from 25 patients with POAG and 25 controls; 14 of the patients presented the GSTM1 gene null polymorphism while only eight of the control group had this gene.Although the POAG patients had a higher frequency of GSTM1, the difference was not significant (P = 0.0874); this lack of significance could be due to the small sample size.

Isolation and characterization of farm pig adipose tissue-derived mesenchymal stromal/stem cells.

Adipose tissue-derived mesenchymal stromal/stem cells (ASCs) are considered important tools in regenerative medicine and are being tested in several clinical studies. Porcine models are frequently used to obtain adipose tissue, due to the abundance of material and because they have immunological and physiological similarities with humans. However, it is essential to understand the effects and safe application of ASCs from pigs (pASCs) as an alternative therapy for diseases. Although minipigs are easy-to-handle animals that require less food and space, acquiring and maintaining them in a bioterium can be costly. Thus, we present a protocol for the isolation and proliferation of ASCs isolated from adipose tissue of farm pigs. Adipose tissue samples were extracted from the abdominal region of the animals. Because the pigs were not raised in a controlled environment, such as a bioterium, it was necessary to carry out rigorous procedures for disinfection. After this procedure, cells were isolated by mechanical dissociation and enzymatic digestion. A proliferation curve was performed and used to calculate the doubling time of the population. The characterization of pASCs was performed by immunophenotyping and cell differentiation in osteogenic and adipogenic lineages. The described method was efficient for the isolation and cultivation of pASCs, maintaining cellular attributes, such as surface antigens and multipotential differentiation during in vitro proliferation. This protocol presents the isolation and cultivation of ASCs from farm pig as an alternative for the isolation and cultivation of ASCs from minipigs, which require strictly controlled maintenance conditions and a more expensive process.

Optimization of silver-containing bioglass nanoparticles envisaging biomedical applications.

Bioglass nanoparticles (BGs) are of outmost importance in the biomedical field, because their unique characteristics, namely osteoconductivity and osteoinductivity, and also in certain conditions, angiogenic and bactericidal properties. In this work, novel bioglass nanoparticles containing silver (AgBGs) were synthesized by a sol-gel method, adopting different thermal treatments to obtain new nanoparticles with bioactive and antibacterial features. This is the first systematic study of the effect of the thermal treatment on the properties of AgBGs. The effect of the studied thermal treatments on the properties of synthesized nanoparticles was analyzed by several characterization techniques: FT-IR, XRD, S-TEM, SEM-EDS and Zeta potential. FT-IR allowed the identification of the characteristic peaks of the nanoparticles and XRD revealed the presence of the characteristic peaks of an apatite-like phase. By S-TEM analysis it was found that the produced nanoparticles are dense and have a diameter <200 nm. The SEM micrographs showed their surface morphology and Zeta potential measurements were performed to study their suspension stability. Additionally, in vitro bioactivity tests confirmed their bioactive potential and the microbiological tests evidenced their bactericidal effect. These promising AgBGs could be incorporated either in 2D or 3D structures for several biomedical applications, namely in the orthopedic and dental fields.

Flaxseed improves embryo production in Boer goats.

Flaxseed is a source of polyunsaturated fatty acids and could be used as a dietary ingredient to enhance reproductive performance of ruminants. The objectives of this study were to determine the effect of feeding diets with different levels of flaxseed on the nutrient intake, and quantity and quality of embryos in Boer goats. A total of 24 multiparous Boer goats were fed with a diet containing either 0, 4, 8 or 12% of flaxseed (n = 6 per group) and subjected to superovulation to determine the quantity and quality of embryos collected on 7 d after natural service. The nutrient intake was linearly associated with levels of flaxseed in the diet and, whereas while the fat (measured as ether extract) intake was positively associated, the non-fiber carbohydrate intake had a negative association with increasing levels of flaxseed in the diet. The quantity, quality and stage of embryonic development on 7 d after natural service were significantly different between levels of flaxseed in the diet. The number of viable embryos was greater in goats fed with a diet containing 4, 8, and 12% flaxseed (94, 84, and 87%, respectively) than those fed with a diet containing 0% flaxseed (65%). On the other hand, the number of degenerated embryos was greater for goats fed with a diet containing 0% flaxseed (35%) than those fed with a diet containing 4, 8, and 12% flaxseed (6, 16, and 13%, respectively). The proportion of grade 1 embryo collected was greater for goats fed with a diet containing 4 and 8% flaxseed (74 and 83%, respectively) than those fed with a diet containing 0 and 12% flaxseed (40 and 46%, respectively). In summary, our study demonstrated that feeding a diet with moderate levels of flaxseed could produce a greater number of better-quality embryos in Boer goats.

Fluoranthene Biodegradation by Serratia sp. AC-11 Immobilized into Chitosan Beads.

The intensive production of polycyclic aromatic hydrocarbons by anthropogenic activities is a serious environmental problem. Therefore, new bioremediation methods are required to avoid widespread contamination. In this work, Serratia sp. AC-11 strain isolated from a tropical peat was selected for immobilization into chitosan beads, which were employed in the biodegradation of fluoranthene. The sizes of the produced beads were relatively uniform with an average diameter of 3 mm. The material was characterized by SEM and FT-IR, confirming the cells immobilization and the protective barrier formed by the chitosan surrounding the biomass. The immobilized bacteria were able to degrade 56% of fluoranthene (the initial concentration was 100 mg L-1) in just 1 day at twice the degradation rate achieved by free-living cells. Furthermore, the immobilized bacteria showed excellent removal during five reuse cycles, from 76% to 59% of biodegradation. These results showed the potential of this approach for remediation of contaminated sites.

Morphological changes in rat rectus abdominis muscle induced by diabetes and pregnancy.

The urethral muscle of diabetic pregnant rats is affected by long-term mild diabetes and short-term severe diabetes, which plays a crucial role in the pathogenesis of pelvic floor disorders. We hypothesized that muscles outside the pelvis are subject to similar changes. The current study aimed at analyzing the effects of long-term mild and short-term severe diabetes on the structure and ultrastructure of fiber muscles and collagen in rats' rectus abdominis (RA) muscle. Therefore, the RA muscle of virgin, pregnant, long-term mild diabetic, short-term severe diabetic, long-term mild diabetic pregnant and short-term severe diabetic pregnant 3-month-old Wistar rats were collected. The structure was analyzed by picrosirius red staining, immunohistochemistry for fast and slow muscle fibers and transmission electron microscopy. We investigated two levels of STZ- induced diabetes: long-term mild diabetes (blood glucose level: 120-200 mg/dL) and short-term severe diabetes (blood glucose level >300 mg/dL). Long-term mild diabetic pregnant and short-term severe diabetic pregnant rats had decreased fast fibers and increased slow fibers, disrupted areas of sarcomere, intermyofibrillar mitochondria and myelin figures in the RA muscle. Both groups enabled us to analyze the specific influence of pregnancy, separately from diabetes. The current study demonstrated that diabetes and pregnancy induced intramuscular transformation and reorganization of RA muscle with a switch of fiber type adjusting their architecture according to intensity and duration of hyperglycemic insult within pregnancy.

Antifungal activity of Brazilian medicinal plants involved in popular treatment of mycoses.

A survey of medicinal plants used to treat common mycoses was done in the Curituba district, Sergipe State, Brazil. One hundred inhabitants were interviewed by health agents and traditional healers. Four different plants were the most cited (more than 50% of the citations): Ziziphus joazeiro, Caesalpinia pyramidalis, Bumelia sartorum and Hymenea courbaril. The aqueous extracts obtained following traditional methods and using different parts of these plants, were submitted to drop agar diffusion tests for primary antimicrobial screening. Only the water infusion extract of Ziziphus joazeiro and Caesalpinea pyramidalis presented a significant antifungal activity against Trichophyton rubrum, Candida guilliermondii, Candida albicans, Cryptococcus neoformans and Fonsecaea pedrosoi, when compared to the antifungal agent amphotericin B. The minimal inhibitory concentration (MIC) of the bioactive extracts was evaluated by the microdilution method. Best activity with a MIC of 6.5 microg/ml for both extracts was observed against Trichophyton rubrum and Candida guilliermondii. Ziziphus joazeiro and Caesalpinea pyramidalis extracts presented also low acute toxicity in murine models. The present study validates the folk use of these plant extracts and indicates that they can be effective potential candidates for the development of new strategies to treat fungal infections.

Isolation and characterization of farm pig adipose tissue-derived mesenchymal stromal/stem cells

Adipose tissue-derived mesenchymal stromal/stem cells (ASCs) are considered important tools in regenerative medicine and are being tested in several clinical studies. Porcine models are frequently used to obtain adipose tissue, due to the abundance of material and because they have immunological and physiological similarities with humans. However, it is essential to understand the effects and safe application of ASCs from pigs (pASCs) as an alternative therapy for diseases. Although minipigs are easy-to-handle animals that require less food and space, acquiring and maintaining them in a bioterium can be costly. Thus, we present a protocol for the isolation and proliferation of ASCs isolated from adipose tissue of farm pigs. Adipose tissue samples were extracted from the abdominal region of the animals. Because the pigs were not raised in a controlled environment, such as a bioterium, it was necessary to carry out rigorous procedures for disinfection. After this procedure, cells were isolated by mechanical dissociation and enzymatic digestion. A proliferation curve was performed and used to calculate the doubling time of the population. The characterization of pASCs was performed by immunophenotyping and cell differentiation in osteogenic and adipogenic lineages. The described method was efficient for the isolation and cultivation of pASCs, maintaining cellular attributes, such as surface antigens and multipotential differentiation during in vitro proliferation. This protocol presents the isolation and cultivation of ASCs from farm pig as an alternative for the isolation and cultivation of ASCs from minipigs, which require strictly controlled maintenance conditions and a more expensive process.

Effect of replacing ground corn and soybean meal with licuri cake on the performance, digestibility, nitrogen metabolism and ingestive behavior in lactating dairy cows.

Licuri (Syagrus coronate) cake is a biodiesel by-product used in ruminant feed as a beneficial energy source for supplementation in managed pastures. The objective was to evaluate the performance, digestibility, nitrogen balance, blood metabolites, ingestive behavior and diet profitability of eight crossbred Holstein (3/4)×Gyr (5/8) multiparous cows (480±25 kg BW and 100 days milking) grazing and supplemented with licuri cake partially replacing ground corn and soybean meal in concentrate (0, 200, 400 and 600 g/kg in dry matter (DM)), distributed in an experimental duplicated 4×4 Latin square design. Licuri cake partially replacing ground corn and soybean meal increased (P<0.01) the intake and digestibility of ether extract and decreased the non-fiber carbohydrates; however, there were no influences on the intakes of DM, CP, NDF and total digestible nutrients (TDN). The digestibilities of DM, CP and NDF were not influenced by licuri cake addition. There was a decrease trend on TDN digestibility (P=0.08). Licuri cake replacing ground corn and soybean meal in concentrate did not affect the intake; fecal, urinary and mammary excretions; N balance; and triglycerides concentrations. However, the blood urea nitrogen (P=0.04) concentration decreased with the licuri cakes inclusion in cow supplementation. There was an increasing trend for serum creatinine (P=0.07). Licuri cake inclusion did not affect body condition score, production, yield, protein, lactose, total solids and solid non-fat contents of milk and Minas frescal cheese. There was a linear decrease in average daily weight gain (g/day). The milk fat concentration and cheese fat production (P<0.1) presented a linear increase with partial replacement of ground corn and soybean meal with licuri cakes. The addition of licuri cake did not alter the time spent feeding, ruminating or idling. There was an increasing trend in NDF feeding efficiency (P=0.09). The replacing of ground corn and soybean meal with licuri cake up to 600 g/kg decreased the concentrate cost by US$0.45/cow per day. Licuri cake replacing corn and soybeans (400 g/kg) in concentrate promoted a profit of US$0.07/animal per day. Licuri cake is indicated to concentrate the supplementation of dairy cows with average productions of 10 kg/day at levels up to 400 g/kg in the concentrate supplement because it provides an additional profit of US$0.07/animal per day and increased milk and Minas frescal cheese fat without negative effects on productive parameters.