Scielo RSS <![CDATA[Brazilian Archives of Biology and Technology]]> http://www.scielo.br/rss.php?pid=1516-891320160001&lang=pt vol. 59 num. lang. pt <![CDATA[SciELO Logo]]> http://www.scielo.br/img/en/fbpelogp.gif http://www.scielo.br <![CDATA[Use of Solid Waste from Thermoelectric Plants for the Cultivation of Microalgae]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100200&lng=pt&nrm=iso&tlng=pt ABSTRACT The aim of this study was to analyze the influence of solid waste on the cultivation of the microalgae Spirulina sp. LEB 18 and Chlorella fusca LEB 111 with 0, 40, 80 and 120 ppm of mineral coal ash. The addition of the ash did not inhibit the cultivation of microalgae at the tested concentrations, showing that it could be used for the cultivation of these microalgae due to the minerals present in the ash, which might substitute the nutrients needed for their growth. <![CDATA[Effects of Sodium Nitrate and Mixotrophic Culture on Biomass and Lipid Production in Hypersaline Microalgae Dunaliella Viridis Teod]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100201&lng=pt&nrm=iso&tlng=pt To access the potential application of Dunaliella viridis Teod. for biofuel production, the effects of culture media composition on biomass and lipid content of this microalgae were investigated. Measured at the 20 th day, sodium nitrate at 5.0 mM augmented biomass production by 26.5 percent compared to control (1 mM sodium nitrate). Total lipids expressed as µg mL-1 of culture also increased with increase in nitrate concentration up to 5.0 mM sodium nitrate, whereas when expressed on the per cell basis, total lipids stayed relatively constant at most of the tested nitrate concentrations except at 0.5 mM which was 31.4 percent higher compared to 1.0 mM nitrate. At 5.0 mM sodium nitrate, by using 20 g L-1 of glucose in mixotrophic culture of D. viridis, cell number augmented by 36.4 percent compared to the cultures with no added glucose. Llipid content per cell and per mL of culture was increased by 71.4 and 135.1 percent, respectively. Among plant hormones, 10-9 M indole-3- acetic acid (IAA) plus 10 -8 M trans-zeatin riboside led to 22.8 percent higher biomass relative to control (without hormone and at 1.0 mM sodium nitrate). It is concluded that altering the growth conditions of D. viridis can lead to higher cell densities and higher lipids content which can be exploited for biofuel production. <![CDATA[Spore Germination and Protonemal Development of <strong><em>Dolichomitriopsis diversiformis</em></strong>]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100202&lng=pt&nrm=iso&tlng=pt ABSTRACT Dolichomitriopsis diversiformis is an endemic and threatened moss in eastern Asia. In vitro culture and light microscopic observation were employed to study its developmental process from spore germination to the formation of young gametophyte, as well as effects of light and temperature on its spore germination and protonemal development. Microscopic observations revealed that its spores were positively photoblastic with exosporous germination, and sporelings were classified as the bryum-type. Light and dark conditions were compared to understand their effects on spore germination. In 24 h continuous illumination, all spores germinated in both 20 ℃ and 25 ℃. In contrast, in darkness at 20 ℃ from one to 30 days, spores did not germinate. However, when dark-cultured spores transferred to continuous light, they started to geminate in 48 hours. In addition, effects of 20 ℃ and 25 ℃ on sporeling were compared. In a continuous light condition, sporelings grew longer and developed more protonema branches in the 20 ℃ treatment than in the 25 ℃ treatment. It was interesting that a phenomenon of slow and hysteretic spore germination and protonematal development was observed in natural light and room temperature (5-10℃). These observations suggest that spore germination and sporeling development is regulated by light and temperature. <![CDATA[Chemical Composition and Antioxidant Activity of Essential Oil and Organic Extracts of <strong><em>Premna integrifolia</em></strong> Linn]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100203&lng=pt&nrm=iso&tlng=pt ABSTRACT This study was designed to examine the chemical composition and in vitro antioxidant activity of the hydrodistillated essential oil and various extracts obtained from Premna integrifolia Linn. GC-MS analysis of the essential oil was resulted in determination 29 different compounds, representing 95.73% of total oil. Antioxidant activities of the essential oil and organic extracts of chloroform, ethyl acetate and methanol were determined by three different test systems namely DPPH (2,2-diphenyl-1-picrylhydrazyl), superoxide and nitric oxide radical scavenging assays. The essential oil and methanol extract showed potent antioxidant activity among all the tested samples. Furthermore, the amount of total phenolic compounds was determined and its content in methanol extract was the highest as compared to other samples. The results indicate that the essential oil and extracts of Premna integrifolia could serve as an important bio-resource of antioxidants for using in the pharmaceutical industries. <![CDATA[Identification and characterization of <strong><em>Fusarium mangiferae</em></strong> as pathogen of mango malformation in India]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100204&lng=pt&nrm=iso&tlng=pt ABSTRACT Fusarium mangiferae (=F. subglutinans) isolates collect from malformed samples from major mango-growing area of North India. Molecular identification and characterization of eleven most virulent isolates of F. mangiferae, based on pathogenicity tests used for the present study. Species-specific, genus specific ITS-PCR and PCR-RFLP performed for the accurate and easy detection of F. mangiferae. The rDNA-ITS 28S region sequences used for phylogenetic analysis of Fusarium isolates from India and other countries for homology search between them. The phylogenetic tree divided the isolates into three clades (i.e., American, Asian and African) and showed the high level of sequence based similarity (69-99%) among all Fusarium sequences from Asia. Thus, claimed Fusarium mangiferae as dominant pathogen of mango malformation. Furthermore, we conclude that exploiting the nested PCR coupled with PCR-RFLP will help in rapid and accurate detection of F. mangiferae pathogen of mango malformation. <![CDATA[Phytotoxicity of Perlatolic Acid and Derivatives]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100205&lng=pt&nrm=iso&tlng=pt ABSTRACT Perlatolic acid (1), methyl perlatolate (2), and the products of perlatolic acid alcoholysis-namely, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, and n-hexyl 2-hydroxy-4-methoxy-6-n-pentylbenzoate (3-11)-were evaluated for their herbicidal potential on Lactuca sativa L. (lettuce) and Allium cepa L. (onion) seeds. The compounds exhibited low phytotoxicity on L. sativa germination. Perlatolic acid (1) proved the most active compound (20%). Activity was lowest for n-hexyl 2-hydroxy-4-methoxy-6-n-pentylbenzoate (11) (3.5%). The esters iso-propyl and sec-butyl 2-hydroxy-4-methoxy-6-n-pentylbenzoate (6 and 8, respectively) inhibited root (35% e 43%, respectively) and hypocotyl growth (59% e 56%, respectively). The esters n-butyl, n-pentyl, and n-hexyl 2-hydroxy-4-methoxy-6-n-pentylbenzoate (7, 10, and 11, respectively) proved phytotoxic to A. cepa, delaying and reducing seed germination (27%), while n-pentyl 2-hydroxy-4-methoxy-6-n-pentylbenzoate (10) was the most inhibitory for root (42%) and coleoptile growth (24%). The behaviors of iso-propyl, sec-butyl, and n-pentyl 2-hydroxy-4-methoxy-6-n-pentylbenzoate (6, 8, and 10, respectively) suggest the potential utility of these esters as natural herbicides. The esters iso-propyl and sec-butyl 2-hydroxy-4-methoxy-6-n-pentylbenzoate (6 and 8, respectively) may serve as model molecules in the investigation of potential herbicides for dicotyledon control, while n-pentyl 2-hydroxy-4-methoxy-6-n-pentylbenzoate (10) may serve the same function for monocotyledon species. <![CDATA[Expression of Ghrelin and GHSR-1a in Long Term Diabetic Rat's Kidney]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100300&lng=pt&nrm=iso&tlng=pt The aim of this work was to study the relative ghrelin and growth hormone secretagogue receptor (GHS-R)1a gene expression in the kidney of long-term diabetic rats. Forty male Wistar albino rats were divided into four groups: C- control group, DI- one month diabetic rats group, DII- two months diabetic rats group, and DIII- three months diabetic rats group. Diabetes was induced by streptozotocin STZ (40mg/kg i.p). The rats were decapitated under ketamine anesthesia and their kidney tissues were removed. Tissue GHS-R mRNA levels, ghrelin expression, and histopathological damage scores were compared. Dilatation in the distal tubules, epithelial desquamation into the lumen of the tubules and transparent tubules showing glycogen vacuolation were observed in all the diabetic groups. Ghrelin immunoreactivity was significantly higher in group DI compared to group C, whereas in groups DII and DIII, ghrelin immunoreactivity was similar with group C. GHSR-1a mRNA level in group DIII was significantly lower than in group C. As a result, ghrelin immunoreactivity increased at the beginning of diabetes; however, with increase in the duration of diabetes ghrelin immunoreactivity approached to the control values. The expression of GHSR-1a mRNA decreased with increase in diabetes duration. It seemed that down-regulation of GHSR-1a contributed to the renal damage induced by long-term diabetes. <![CDATA[Viability of Human Gingival Fibroblast (FGH) Treated with Ethanolic "Aroeira" Extract (Myracrodruon urundeuva Allemão)]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100301&lng=pt&nrm=iso&tlng=pt ABSTRACT The aim of this study was to evaluate the effect of ethanolic "aroeira" (Myracrodruon urundeuva) extract on the viability of human gingival fibroblast. For this, fibroblasts (2x103 cells/well) were plated in a 96-well plate and incubated for 24 h; the medium (Eagle's medium modified by Dulbecco - DMEM) supplemented with 10% fetal bovine serum was replaced by DMEM with different ethanolic extract concentration (0, 0.1, 1, 10, 100, and 1000μg / mL). The fibroblast viability was analyzed after 48,72, and 96 h by the neutral red capture test and violet crystal. The "aroeira" extract, at high concentrations (100 and 1000 µg/mL) caused decrease in both cellular viability tests (p&lt;0.05). However, dilutions between 0.1 and 10 µg/mL did not affect the viability of the cells. It was concluded that "aroeira" extract was able to change the gingival fibroblast viability, and this effect was concentration dependent. <![CDATA[Isolation, Culturing, Characterization and Aging of Adipose Tissue-derived Mesenchymal Stem Cells: A Brief Overview]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100302&lng=pt&nrm=iso&tlng=pt ABSTRACT The aim of this review was to describe the current state-of-the-art regarding isolation, characterization and aging of adipose tissue-derived mesenchymal stem cells (ADSCs). Mesenchymal stem cells (MSCs) have recently received widespread attention because of their potential use in tissue-engineering applications. Various studies have indicated that MSCs with a fibroblast-like morphology migrate to the sites of injury and help to regenerate damaged tissue. Over the past few years, it has been recognized that fat is not only an energy supply, but also a rich source of multipotent stem cells that can be easily harvested, isolated and selected as compared with other tissues. ADSCs are particularly interesting because of their rapid proliferation and multidirectional differentiation potential. <![CDATA[Liposome and Their Applications in Cancer Therapy]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100303&lng=pt&nrm=iso&tlng=pt ABSTRACT Liposomes, the vesicles of phospholipid bilayer, can encapsulate both hydrophilic and lipophilic drugs and protect them from degradation. Liposomes have been extensively studied and continue to create intense interest in research since their discovery in the mid-1960s. Since then, liposomes have been considered to be the most successful nanocarriers for drug deliver and have made their way to the market. Currently, a number of liposomal formulations are on the marker for cancer treatment and many more are in pipe line. This review discusses about the liposome components, methods of preparation, drug encapsulation mechanism and the potential therapeutic applications of liposomes in cancer therapy. <![CDATA[Helicobacter pylori Infection is a Significant Factor Risk for Hyperhomocysteinemia in the Patients with Coronary Artery Disease]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100304&lng=pt&nrm=iso&tlng=pt ABSTRACT This work aimed to determine whether seropositivity to Helicobacter pylori infection was an independent risk factor for hyperhomocysteinemia patients with cardiovascular disease. The H. pylori IgG, IgA and homocystein levels in 96 patients with cardiovascular disease and 64 participants free of cardiovascular disease as control subjects were determined by ELISA assay. The results showed that seropositivity to H. pylori IgG and IgA levels of coronary artery disease (CAD)patients was significantly higher than the controls and CAD patients with H. pylori IgG and IgA negative antibodies. A significant correlation was found between the seropositivity to H. pylori IgG and homocysteine levels of CAD patients in comparison with the controls and CAD patients with seronegativity to H. pylori IgG and IgA (r=0.233, P= 0.019 ). The involvement of H. pylori infection in atherosclerosis process was based on the chronic inflammation, which might facilitate the CAD-related pathologies. The effect of the presence of H. pylori infection on homocysteine levels elevation in the CAD patients (as a risk factor independent of other traditional factors) was remarkable. <![CDATA[Impact of Equine Chorionic Gonadotropin Associated with Temporary Weaning, Estradiol Benzoate, or Estradiol Cypionate on Timed Artificial Insemination in Primiparous Bos Indicus Cows]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100305&lng=pt&nrm=iso&tlng=pt The study aimed to determine the impact of equine chorionic gonadotropin (eCG) associated with different timed artificial insemination (TAI) protocols on the pregnancy rate (PR) in Bos indicus cows previously treated with progesterone. Five hundred and fifty-seven primiparous cows were subjected to the following treatments: on day 0 (d0), GeCGTW (group equine Chorionic Gonadotropin+Temporary Weaning;n=178) received 0,558 g intravaginal progesterone (P4)+1.0 mg of estradiol benzoate (EB) (IM); on d8 (P4 removal+0,075 mg D-cloprostenol + 400 IU eCG + TW for 48 h); on d10, TAI + calves return to dam; GeCGEB (group equine Chorionic Gonadotropin+Estradiol benzoate; n=176) the same as GeCGTW without TW + application of 1.0 mg of EB on d9; GeCGEC (group equine Chorionic Gonadotropin+Estradiol Cypionate; n=203), the same as GeCGTW without TW+1.5 mg EC (IM). On d35, post TAI, pregnancy diagnosis (PD) was performed. Non-pregnant animals remained under clean-up bulls for 90 days. After this period, the animals were subjected to PD using ultrasound. The PR of TAI was 51.1%, 47.1%, and 47.8% for GeCGTW, GeCGEB24, and GeCGEC (P&gt;0.05) respectively. The PR under clean-up bulls was 88.3%, 47.3%, and 31.1% (P&lt;0.05). The final PR (TAI+clean-up bulls) of the groups was 94.4%, 72.1%, and 64.0%, respectively (P&lt;0.05). It was concluded that no differences in PR among the protocols related to TAI were detected; PR in the GeCGTW protocol under clean-up bulls was higher compared to others (P&lt;0.05); the overall PR of cows subjected to TAI+clean-up bulls was significantly higher in GeCGTW than in the other groups. <![CDATA[Effect of LPS on the Viability and Proliferation of Human Oral and Esophageal Cancer Cell Lines]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100306&lng=pt&nrm=iso&tlng=pt The esophagus and mouth tumors are very frequent malignancies worldwide. Lipopolysaccharides (LPS) are capable of regulating gene expression of pro-inflammatory cytokines by binding to toll-like receptor 4 (TLR4). Recent studies show that LPS can increase the migration ability of human esophageal cancer cell line HKESC-2 by increasing its adhesion properties. However, the effect of LPS has not been tested on viability of human esophageal and oral cancer cells. This study aimed to determine the action of LPS on the cell proliferation and viability in OE19 (adenocarcinoma) and OE21 (squamous carcinoma) cell lines, representative of human esophageal cancer, and HN30 cell line, representative of human oral carcinoma. LPS was used as treatment to OE19 and OE21 cells, and PgLPS (Porphyromonasgingivalis lipopolysaccharide) to HN30 cells. Viability was assessed by MTT assay and proliferation by cell counting. TLR4 expression was evaluated by real-time PCR. LPS at higher concentrations decreased significantly cell viability in both cell lines, adenocarcinoma (OE19) and squamous esophageal carcinoma (OE21) at different times of treatment. In addition, both cell lines, OE19 and OE21, expressed TLR4 receptor. Taken together, our data demonstrated that LPS at high concentrations might contribute to tumor death, in agreement with previously data. <![CDATA[Cytotoxic Effect on Cancerous Cell Lines by Biologically Synthesized Silver Nanoparticles]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100307&lng=pt&nrm=iso&tlng=pt The biosynthesis of nanoparticles has been proposed as an environmental friendly and cost effective alternative to chemical and physical methods. Silver nanoparticles are biologically synthesized and characterized were used in the study. The invitro cytotoxic effect of biologically synthesized silver nanoparticles against MCF-7 cancer cell lines were assessed. The cytotoxic effects of the silver nanoparticles could significantly inhibited MCF-7 cancer cell lines proliferation in a time and concentration-dependent manner by MTT assay. Acridine orange, ethidium bromide (AO/EB) dual staining, caspase-3 and DNA fragmentation assays were carried out using various concentrations of silver nanoparticles ranging from 1 to 100 μg/mL. At 100 μg/mL concentration, the silver nanoparticles exhibited significant cytotoxic effects and the apoptotic features were confirmed through caspase-3 activation and DNA fragmentation assays. Western blot analysis has revealed that nanoparticle was able to induce cytochrome c release from the mitochondria, which was initiated by the inhibition of Bcl-2 and activation of Bax. Thus, the results of the present study indicate that biologically synthesized silver nanoparticles might be used to treat breast cancer. The present studies suggest that these nanoparticles could be a new potential adjuvant chemotherapeutic and chemo preventive agent against cytotoxic cells. However, it necessitates clinical studies to ascertain their potential as anticancer agents. <![CDATA[Comparison of therapeutic effects of L-Thyroxin, apelin and a combination of both on antioxidant enzymes in the heart of PTU-induced hypothyroid rats]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100308&lng=pt&nrm=iso&tlng=pt Atherosclerosis is one of the common disorders among hypothyroidism, which, increased the risk of cardiovascular diseases. Reactive oxygen species are associated with atherosclerosis development. Antioxidant defense systems are the scavenger for free radicals. Apelin is an endogenous ligand for the APJ receptor (apelin receptor) that exists in most tissues, acts as an adiponectin. It has been identified that apelin administration, improve the antioxidant capacity (TAC). Therefore, this study was conducted to assess, therapeutic effects of apelin, T4 (L-Thyroxin) or both on antioxidant capacity in 6-propyl-2-thiouracil (PTU)-induced hypothyroid rats. Forty male Wistar rats were randomly assigned into five groups: C: control group; P group (hypothyroid): PTU (0.05 %) administration for six weeks; P+A, P+T and P+A+T groups: after 4 weeks of PTU administration, animals treated with Apelin (200 μg/kg/day, ip) T4 (0.02 µg/g/day, gavage) and apelin+T4; for two weeks respectively accompanied by PTU administration. Aplein administration in P+A group and P+A+T group had beneficial effect to lowering of malondialdehyde (MDA) content as compared to hypothyroid group (8.52±0.64 and 8.53±1 vs. 13.67±1.64 nmol/g tissue, P&lt;0.05) and also had increasing effect on Superoxide dismutase (SOD) and glutathion peroxidase (GPx) activity and the total antioxidant capacity (TAC) content compared to the hypothyroid group. This study showed that apelin was able to improve the oxidant-antioxidant balance in the heart tissue of the hypothyroid rats by elevating of antioxidant enzyme activity. <![CDATA[Nuclear translocation of STAT3 by in vitro metreleptin administration causes lipolysis in human primary adipocytes]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100309&lng=pt&nrm=iso&tlng=pt We utilized subcutaneous (SC)- and omental (OM)-derived human primary adipocytes (hPA) from obese male, and investigated whether synthetic analog of leptin, metreleptin, may regulate lipolysis via translocation of STAT3 to the nucleus. We observed that 50 ng/mL of metreleptin increases STAT3 phosphorylation in both SC- and OM-derived hPA. Importantly, we found for the first time that metreleptin is capable of trans-locating STAT3 to the nucleus and STAT3 blockade inhibits metreleptin-induced lipolysis. Our initial data provide novel insights into the role of STAT3 as probable mediator of the action of metreleptin in regulating metabolism. <![CDATA[Regulating Effect Of Carnosine And /Or L- Arginine On The Expression Of Inflammatory Molecules Induced Nephropathy In The Hypoxic Rat Model]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100310&lng=pt&nrm=iso&tlng=pt This study aimed to explore the effective role of carnosine and /or L- arginine in down regulation of the inflammatory molecule expression caused renal damage in response to sodium nitrite (NaNO2) induced hypoxia in rats . NaNO2 was administered subcutaneously (s.c.) to rats as a single dose (60 mg/kg body weight ). L-arginine (200mg/Kg body weight) and carnosine (250 mg/ Kg body weight ) were administered (i.p.) as a single dose , 24 h before NaNO2 injection. The results revealed that pre- administration of arginine and /or carnosine to NaNO2 hypoxic rats, significantly modulated the increases in serum markers of renal function (creatinine and urea) as well as the decrease in hemoglobin (Hb) level versus hypoxic rats. The two agents each alone or in a combination, markedly down regulated the serum pro-inflammatory molecules, including tumor necrosis factor-α (TNF- α) , C-reactive protein (CRP), vascular endothelial growth factor (VEGF) and heat shock protein -70 (HSP-70) as well as interleukin-6 (IL-6) in renal tissue compared to NaNO2 hypoxic rats . Also, the two agents successfully down modulated the alteration in the serum hypoxia inducible factor 1α (HIF 1α) . The present biochemical results were also supported by histopathological examination. In conclusion, the current data revealed that although the efficacy of arginine or carnosine each alone, their combination was more effective in ameliorating the renal damage induced by inflammatory molecules in response to NaNO2 hypoxia . This may support the use of this combination as an effective drug to treat hypoxic renal damage <![CDATA[Investigation of the <em>GJB6</em> Deletion Mutations Del (GJB6-D13s1830) and Del (GJB6-D13s1854) in Iranian Patients with Autosomal-Recessive Non-Syndromic Hearing Loss (ARNSHL)]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100311&lng=pt&nrm=iso&tlng=pt Hearing loss (HL) is the most common inherited sensory disorder affecting about 1 in 1000 births. The first locus for nonsyndromic autosomal recessive HL is on chromosome 13q11-22. The two genes, GJB2 and GJB6, are closely located on chromosome and are known to be co-expressed in the embryonic cochlea. Deletion mutations involving GJB6 were associated with autosomal-recessive nonsyndromic hearing loss (ARNSHL) and in combination with a GJB2 mutation with digenic ARNSHL. The objective of this study was to screen for the del (GJB6-D13S1830) and del (GJB6-D13s1854) mutations in GJB6 gene in patients with ARNSHL from Iran, using multiplex PCR and direct sequencing methods. Agarose gel electrophoresis and DNA sequencing of amplified fragment of the PCR reaction showed none of the patients was found to carry deletion in GJB6 gene which indicates that these deletions are restricted to certain populations and indicating a founder effect regarding these deletions. <![CDATA[Antioxidant Activity of Oral Administration of Rosmarinus Officinalis Leaves Extract on Rat's Hippocampus which Exposed to 6-Hydroxydopamine]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100312&lng=pt&nrm=iso&tlng=pt Carnosic acid, a diterpene of Rosemarinus officinalis leaves extract (RE), has potent antioxidant activity in vitro. The dopaminergic connection of substantia nigra pars compacta to the hippocampus might be affected by oxidative stress which caused cognitive impairment observed in the early phase of Parkinson's disease (PD). Adult male Wistar rats were lesioned bilaterally by intra-nigral injection of 6-OHDA, and divided into six groups: four groups that orally given RE containing 40% of carnosic acid, at doses of 25, 50 and 100 mg/kg (treated rats) and distilled water (H2O), once daily for a period of 14 days before and after the injury. There were also two another groups as control rats which injected by normal saline and untreated lesion group. The injured animals were evaluated for their spatial memory performance by Morris Water Maze test. Lesioned rats showed significant increase in escape latency, as compared with control group. Two weeks after injury, tissue samples were collected from the hippocampus. Levels of catalase (CAT), glutathione peroxidase (GPX) and superoxide dismutase (SOD), malondialdehyde (MDA) and reactive oxygen species (ROS) were determined. There were significant increase of SOD, GPX and CAT enzymes activities in RE50 treated group as compared to lesioned rats. We found a significant decrease of ROS in RE50 treated group as compared to Lesioned rats. These findings provide evidence that 50 mg/kg of RE decreased oxidative damage of the hippocampus induced by 6-OHDA and serve as potential candidate for the treatment of PD. <![CDATA[Effects of Vitamin K1 Supplementation on the Risk Factors to the Stroke and on Memory in Spontaneously Hypertensive Rats Stroke Prone (SHR-sp)]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100313&lng=pt&nrm=iso&tlng=pt A number of risk factors have been associated to the stroke and many strategies have been proposed in order to control them as well. Vitamin K has been largely found in brain, which suggests a possible function at that tissue. This study aimed to evaluate the potential of this vitamin on the prevention of risk factors to stroke and on cognitive function on SHRSP rats. Twelve SHRSP males, 15 weeks old, were divided into two groups (n= 6 each), receiving the vehicle-coconut oil (control group) or 40 μg of phylloquinone (treated group) during 28 days. Biological parameters, systolic blood pressure and lipid profile were evaluated. Both groups were submitted to the neurological tasks. The data was treated by Student's t test and ANOVA one-way test being P&lt;0.05 considered significant. The phylloquinone supplementation showed a statistically significant reduction in the treated group of all parameters of lipid profile and systolic blood pressure when compared to the control group. Neurological evaluation indicated a statistically significant improvement in the performance of long term memory tests in the treated group, without similar findings in the evaluation of short memory. In sum, phylloquinone supplementation was shown to modulated lipid profile and protect neuronal suffering in this model. <![CDATA[Lycopene Protects the Diabetic Rat Kidney Against Oxidative Stress-mediated Oxidative Damage Induced by Furan]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100314&lng=pt&nrm=iso&tlng=pt Furan is a food and environmental contaminant and a potent carcinogen in animals. Lycopene is one dietary carotenoid found in fruits such as tomato, watermelon and grapefruit. The present study was designed to explore the protective effect of lycopene against furan-induced oxidative damage in streptozotocin (STZ)-induced diabetic rat kidney. At the end of the experimental period (28 days), we found that lycopene markedly decreased the malondialdehide (MDA) levels in the kidney, urea, uric acid and creatinine levels in the serum of furan-treated rats. The increase of histopathology in the kidney of furan-treated rats were effectively suppressed by lycopene. Furthermore, lycopene markedly restored superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) activities in the kidney of furan-treated rats. In conclusion, these results suggested that lycopene could protect the rat kidney against furan-induced injury by improving renal function, attenuating histopathologic changes, reducing MDA production and renewing the activities of antioxidant enzymes. <![CDATA[Bromelain: Methods of Extraction, Purification and Therapeutic Applications]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100315&lng=pt&nrm=iso&tlng=pt Bromelain is a concoction of sulfhydryl proteolytic enzymes. Depending upon the site of extraction it can be regarded as either stem bromelain (SBM) (EC 3.4.22.32) or fruit bromelain (FBM) (EC 3.4.22.33). Bromelain remain enzymatic active over a broad spectrumand endure a range of pH (5.5 to 8.0) and temperature (35.5 to 71 ºC). It is one of the extensively investigated proteolytic enzyme owing to its astonishing applications in various industries. This necessitated employing a strategy that result in highest purified bromelain in less steps and lowest cost. Use of modernistic approach such as membrane filtration, reverse micellar systems, aqueous two phase extraction and chromatographic techniques have shown promise in this regard. Besides its industrial applications, bromelain has been widely utilized as a potential phytomedical compound. Some of its reported actions include inhibition of platelet aggregation, anti-edematous, anti-thrombotic, anti-inflammatory, modulation of cytokines and immunity, skin debridement and fibrinolytic activity. It also assist digestion, enhance absorption of other drugs and is a potential postoperatively agent that promote wound healing and reduce postsurgical discomfort and swelling. <![CDATA[Investigation of the Acute Effects of Dry Extract of Glycine Max on Postprandial Glycemia in Rats]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100316&lng=pt&nrm=iso&tlng=pt The acute effects of Glycine max (GM) on post prandial glycemia (PPG) in male Wistar rats were investigated. All substances were orally administered by gavage in overnight fasted animals. The elevation of PPG promoted by starch (1g/kg) was prevented by GM (2.5 mg/kg, 5.0 mg/kg, 7.5 mg/kg, 10.0 mg/kg, and 100.0 mg/kg). In conclusion GM showed potential antidiabetic effect. <![CDATA[Personalized Gene Expression Analyses of SMAD7 and KLF10 In Breast Cancer]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100317&lng=pt&nrm=iso&tlng=pt Many cancer researchers use gene expression analysis for differentiation between tumor and normal cells for diagnostic, prognostic and therapeutic purposes. Most of studies compare either tumor cell lines by normal cell lines or tumor tissue of affected individuals by normal healthy control tissue. But expression of each special gene is unique in different individuals and also in different tissue of same individual. For this reason, here we compare the gene expression levels of SMAD7 and KLF10 in tumor cells and its adjacent normal tissue of breast cancer patients and compared them. For this purpose, a total of 40 tumor and matched tumor-free margin samples were obtained during surgery. The SMAD7 and KLF10 mRNA expression levels in tumor and marginal samples were examined by real-time quantitative PCR. Results are not concordant with previous studies and comparison of only SMAD7 or KLF10 is not useful for differentiating between tumor and margin cells, but ratio analysis of these two genes, SMAD7/ KLF10, can be indicative than study of one gene alone. We concluded that gene expression analysis of tumor cells with adjacent normal tissue are essential for precise identification and interpretation of cancer alterations and have important implications for the diagnostic and therapeutic management of cancer patients. <![CDATA[Antidiabetic Potential of Potentilla fulgens Roots in Validated Animal Models of Diabetes]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100318&lng=pt&nrm=iso&tlng=pt The present study was undertaken to investigate the antidiabetic potential of tap roots of Potentilla fulgens in streptozotocin induced diabetic rat models. The crude powder, ethanolic, ethanolic: aqueous and aqueous extracts of tap roots were administered to normoglycemic- and streptozotocin (STZ)-induced diabetic rats in a single dose study. The ethanolic extract showed significant improvement in oral glucose tolerance and antihyperglycemic effect on sucrose loaded normal rats and STZ-induced diabetic rats. Of the isolated aqueous, n-butanol, chloroform and n-hexane soluble fractions of the active ethanolic extract of the roots, the aqueous fraction (100 mg/kg body weight) showed significant blood glucose lowering effect on STZ-induced diabetic rats. In a multiple dose study, aqueous fraction of ethanolic extract of P. fulgens roots significantly improved the body weight, percent glycated hemoglobin (%HbA1c), fasting blood glucose, oral glucose tolerance (OGTT), serum insulin, lipid profile, liver and kidney parameters in STZ-induced diabetic rats. The aqueous fraction also showed marked improvement in OGTT and serum insulin level in neonatal STZ-induced diabetic rats for 30 consecutive days. The aqueous fraction of the roots also inhibited the activity of alpha (α)-glucosidase enzyme in a dose dependent manner. In conclusion, the finding suggested that an aqueous fraction of tap roots of P. fulgens possessed potential antidiabetic activity. <![CDATA[Potential medicinal use of forest species of the Deciduous Seasonal Forest from Atlantic Forest Biome, South Brazil]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100319&lng=pt&nrm=iso&tlng=pt The current paper focuses on potential medicinal use of forest species from Deciduous Seasonal Forest in central region of Rio Grande do Sul State, South Brazil. Floristic and phytosociological surveys were carried out in 2011 and 2012. Results were compared with the available information in scientific literature about popular knowledge and biological/pharmacological evidences. Each species was classified in use categories to diseases, symptoms our disorders based on the International Statistical Classification of Diseases and Related Health Problems (ICD). In 2011, we found a total of 31 species, 77,4% of them with medicinal use. In 2012, we found 33 species,72,7% of them with medicinal use. The species most frequently mentioned were Casearia sylvestris, Eugenia uniflora, Cabralea canjerana, Luehea divaricata, Parapiptadenia rigida and Vitex megapotamica. The main species with biological/pharmacological properties were C. sylvestris, E. uniflora, L. divaricata, V. megapotamica and Zanthoxylum rhoifolium. We concluded that there is a considerable potential for medicinal purposes to forest species from Deciduous Seasonal Forest in the study region. We recommend further studies mainly about the species Prunus myrtifolia, Zanthoxylum rhoifolium, Cabralea canjerana and Casearia sylvestris which presented references to antitumoral properties. <![CDATA[The Role of Apollon Gene Silencing on Viablity and Radiosensitivity of Cervical Cancer Hela Cells]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100320&lng=pt&nrm=iso&tlng=pt Cervical cancer is the second common cause of cancer deaths in women worldwide. Radioresistancy of cancer is a principal cause of treatment impairing. Inhibitor of apoptosis proteins (IAPs) widely block apoptosis against apoptotic stimuli, including current chemo- and radiation therapies. Apollon, a membrane of IAP, can support cells against apoptosis and is over expressed in some treatment-resistant cancer cells. The aim of this study was to evaluate the effects of apollon knockdown on induction of apoptosis and also its potential for enhancement of radiosensitvity on hela cells. plasmid encoding shRNA which has been confirmed its effect against apollon, transfected into hela cells. Consequent effects on the level of P53 , Bax and BAK analyzed by real time PCR. Apoptotic phenotype of transfected cells was monitored by Tunnel assay. Viability of hela cells after radiotherapy was analyzed by MTT assay. shRNA1 effectively increased transcription of p53, Bax and BAK and induced apoptosis phenotype of treated hela cells. Radiosensitivity of transfected cell was increased after knock-down of apollon obviously. Apollon knockdown induces apoptosis in hela cell . Also it can be as new molecular target for radio-sensitizing strategies in these cells. So, apollon can be a potentially considerable therapeutic object for cervical cancer. <![CDATA[Heregulin-1β Promotes the Synergistic Effect Between Allogenic Skin-Derived Precursor Differentiated Schwann Cells (SKP-SC) and Acellular Nerve Allograft (ANA) in Peripheral Nerve Regeneration Through Inhibiting Mir-21]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100321&lng=pt&nrm=iso&tlng=pt Previous studies in our lab found that heregulin-1β with SKP-SCs (neurons and Schwann cells differentiated from SKPs) / ANA (acellular nerve allograft) transplantation represented a powerful therapeutic approach, and facilitates the efficacy of ANA in peripheral nerve injury. In this study, our purpose is to explore the mechanism between them. Firstly we transplanted ANA + SKP-SC + heregulin-1β into rats with right sciatic nerve injury and then detected the miR-21 and SOX2 (SRY-like HMG box 2) levels. Then we transfected miR-21 inhibitor in SCs (Schwann cells) which induced in hypoxic condition before harvesting. Then we detected expression of miR-21 and SOX2 using real time-PCR and western blot assay. Results in vivo showed that the expression of miR-21 in rats was inhibited after transplantation of ANA + SKP-SC + heregulin-1β with induced SOX2 accordingly. Then we found miR-21 was increased time dependently in hypoxic SCs with decreased SOX2 accordingly. After miR-21 inhibitor transfection, miR-21 level was reduced and SOX2 was up-regulated. Meanwhile it was also showed that the miR-21 inhibitor induced the hypoxic SCs growth, decreased the apoptosis with cell cycle changing. In conclusion miR-21 and its target gene SOX2 played important role in peripheral nerve injury. Heregulin-1β may increase the synergistic effect between SKP-SC and ANA through inhibiting miR-21 in vivo. <![CDATA[Antidiabetic Effect of Oleuropein from Olea europaea Leaf against Alloxan Induced Type 1 Diabetic in Rats]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100322&lng=pt&nrm=iso&tlng=pt This study have been designed to study the effect of extracted pure oleuropein from Oleaeuropaealeaf against alloxan induced type 1 diabetic rats.Diabetic male ratwas induced by injectingsingle subcutaneous injection of 100 mg/kg b.wof alloxan.Respectively, pure oleuropein compound (5, 10, 15 and 20 mg/kg.b.w)was orally administered once per a day during a period of 40 days ofexperiment.Then, the serum blood was collected for the determination of glucose level,haematological analysis, enzymatic and non-enzymatic antioxidant. Further, pancreatic tissue was evaluated for histological examination. Oleuropein showed a significant rolein attenuating the blood glucose levels and elevation of in-vivo antioxidantafter treating diabetic rats with 5 and 10 mg/kg. b.w.The haematologicaltest did not show any significant differences. The histological sections of diabetic rats treated with 5mg/kg/ b.w ofoleuropein showed regularity in size appearance of pancreatic islet with normal distribution of islet cells.Oleuropein as a natural active compound have antioxidant activity to attenuate the effect of alloxan against diabetic disease.Therefore, it can be recommended to use oleuropein as an additive food to cure type 1 diabetic. <![CDATA[Transgenic Expression and Identification of Recombinant Human Proinsulin in Peanut]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100323&lng=pt&nrm=iso&tlng=pt The increased incidence of diabetes, coupled with the introduction of alternative insulin delivery methods that rely on higher doses, is expected to result in a substantial escalation in the future demand for affordable insulin. Plant-based systems offer a safe and economical method for producing pharmaceutical proteins. We used peanut (Arachis hypogaea L.) as bio-reactors to express biosafe, stable proinsulin. We designed two proinsulin analogues (FAIA and LAIA) with substitutions in their amino acid sequences. The fast-acting insulin analogue (FAIA) contains a Gly inserted between Cys19 and Gly20, as well as a Pro28Asp substitution, in the B chain. The long-acting insulin analogue (LAIA) contains a Gly inserted between Cys19 and Gly20 and two Arg residues inserted into the terminus of the B chain, as well as an Asn21Gly substitution in the A chain. Four plasmids were constructed: pROKII-Flag-FAIA, pROKII-Flag-LAIA, pCAMBIA2301-Oleosin-FAIA and pCAMBIA2301-Oleosin-LAIA. These plasmids were transferred into peanut to produce recombinant proinsulin. Western blot and GUS staining analysis indicated that some transgenic peanut successfully expressed exogenous proinsulin. Peanut seeds can act as insulin storage sites, which is the foundation for further production of recombinant proinsulin from peanut seeds. <![CDATA[Osteoarthritis in horses - Part 2: a review of the intra-articular use of corticosteroids as a method of treatment]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100324&lng=pt&nrm=iso&tlng=pt ABSTRACT: The problem considered in this review is related to the frequent use of corticosteroids (COs) (i.e. steroids) in the practice of equine medicine and surgery as a therapeutic strategy in the treatment of degenerative joint disease, commonly known as osteoarthritis (OA). This disease is one of the most common among the equine musculoskeletal diseases, and is clinically characterized by pain, lameness, joint effusion at the initial stage, which usually reduces with the progression of the disease, and reduced physical performance. Although steroids are considered excellent drugs in the control of clinical signs resulting from osteoarthritis, they also influence cellular activity through activation of various signaling mechanisms. However, they can cause adverse effects when administered intra-articularly, since they are immunosuppressive drugs of many cell types. They are also incriminated as suppressors of the chondrocyte matrix synthetic activities, which may contribute to 'arthropathy by corticosteroids', which can also be associated with cumulative injury resulting from improper use (dose and frequency of application) of already damaged joints. The objective of this article is to review information about the advantages and disadvantages of intra-articular COs for treatment of the disease in horses. In addition, some important information of other species is also presented. <![CDATA[Osteoarthritis in horses - Part 1: relationship between clinical and radiographic examination for the diagnosis]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100325&lng=pt&nrm=iso&tlng=pt ABSTRACT Locomotor disorders are very common in the equine clinic, which may be partly due to the different types of activities horses develop. Osteoarthritis (OA), commonly known as degenerative joint disease, presents an considerable role in the series of disorders of the musculoskeletal system and may be associated with other problems such as navicular syndrome, periostitis or osteochondrosis. This affection causes progressive deterioration of articular cartilage, accompanied by bone and soft-tissue periarticular changes. In fact, it results from a complex interaction between biochemical and biomechanical factors. The objective of this article is to review information about clinical and radiographic findings of OA, the biochemical and biomechanical changes manifested in the disease and the importance of the synovial fluid. Additionally, some information on other species is also presented. This review refers to Part 1 of a study whose sequence is entitled "Osteoarthritis in horses - Part 2: a review of the intra-articular use of corticosteroids as a method of treatment." <![CDATA[Analysis of Maceaene and Macamide Contents of Petroleum Ether Extract of Black, Yellow, and Purple <strong><em>Lepidium Meyenii</em></strong> (Maca) and Their Antioxidant Effect on Diabetes Mellitus Rat Model]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100326&lng=pt&nrm=iso&tlng=pt ABSTRACT Maceaene and macamide contents as well as antioxidant effect of petroleum ether extract of black maca (BM), yellow maca (YM), and purple maca (PM) on diabetes mellitus (DM) rats were investigated. The results showed that seven, six, and five analogues of macamides were identified from the petroleum ether extracts of BM, YM, and PM, respectively. BM extract exhibited the highest contents of total macamides. Comparatively, the PM extract has the lowest macamide quantity. The maceaene contents in all the extracts showed no significant difference (p&gt;0.05). Macamide contents in maca with the same color were not statistically different. Pharmacological results showed that 60-day oral administration of the petroleum ether extract of maca (100 mg/kg.d) can significantly decrease lipid oxidation as indicated by the decreased thiobarbituric acid reactive substances (TBARS) and carbonylated proteins (CP) concentrations on DM rat model (P&lt;0.05). Among them, oral administration of PM extract showed the lowest TBRAS and CP concentrations. All maca extracts can enhance antioxidant enzyme (SOD, superoxide dismutase; CAT, catalase) activity of liver and red blood cells (RBC) of DM rat. However, only oral administration of PM extract can increase SOD and CAT activity of both RBC and liver. The glutathion (GSH) contents in plasma were significantly increased in DM rats treated with PM extract (p&lt;0.05). But, oral administration of BM and YM extracts did not enhance GSH levels. Take together, the data suggested that PM extract exhibited the most potent antioxidant activity on DM rat model. And, maceaene and macamide in maca extract was not correlated with its antioxidant ability. <![CDATA[Clotting and fibrinogenolysis inhibition by essential oils from species of the Asteraceae family]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100327&lng=pt&nrm=iso&tlng=pt ABSTRACT Essential oils are pharmacologically active and unexplored compounds. The inhibitory properties of essential oils from Baccharis dracunculifolia, Conyza bonariensis, Tithonia diversifolia and Ambrosia polystachya were evaluated in the coagulation and fibrinogenolysis induced by snake venoms. The essential oil from Conyza bonariensis extended the clotting time of Lachesis muta from 52.2 to 115.2 seconds and that of Bothrops moojeni from 108.3 to 2340.0 seconds, when pre-incubated with the venoms. The longest clotting times for Bothrops atrox venom were observed after incubation with the essential oils from Conyza bonariensis and Tithonia diversifolia: the times increased from 100.8 to 264.0 and 227.7 seconds, respectively. The prior incubation of the essential oils with plasma and subsequent addition of Lachesis muta venom resulted in a pro-clotting effect. The oils from Ambrosia polystachya and Baccharis dracunculifolia caused 100% of inhibition on the fibrinogenolysis induced by Bothrops moojeni and Lachesis muta venoms (the oils were previously incubated with the venom). The results indicate that the essential oils show promise as adjuvants for the treatment of snakebites. <![CDATA[Green Tropical Phytoextracts - Promising Anticancer Alternative]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100328&lng=pt&nrm=iso&tlng=pt ABSTRACT The anticancer potential of aqueous extracts of tropical plants Ficus deltoidea and Labisia pumila were tested on human prostate carcinoma (DU145) cells. The effects of these extracts on the cells were observed through several tests such as cytotoxicity MTT assay, trypan blue exclusion assay (for cell viability), fluorescence and morphological observations. Both of the plant extracts had a growth repression effect on the cells with different concentrations. The best concentration (1x10-3 μg/mL) was chosen by determining their cytotoxic effects on human prostate cancer as well as fibroblast (normal) cells. The F. deltoidea extract showed more potency and killed the cells faster than L. pumila. Morphologically both extracts affected the cells in a similar way, i.e. the cells became irregular shaped, detached themselves and kept floating in the medium. <![CDATA[Insilico Analysis of Phytoconstituents from <strong><em>Allium sativum</em></strong> as Potential Inhibitors of Inha in <strong><em>Mycobacterium tuberculosis</em></strong>]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100329&lng=pt&nrm=iso&tlng=pt ABSTRACT Tuberculosis is leading cause of death among the global bacterial infections. The main causative for tuberculosis is Mycobacterium tuberculosis, which will survive in its host human being for decades in latent or chronic levels. In addition, the late multidrug resistance at a disturbing rate accompanies the appearance of tuberculosis. The quick spread of resistance to initial stage treatment medications has redirected the focus of the medical community in the creation of an array of new drug against Mycobacterium tuberculosis. The InhA protein is a component of Fatty acid synthetase (FAS) II and exhibits an NADH reliant enoyl-ACP reductase activity. InhA is a vital enzyme of M.tuberculosis in control of cell wall synthesis, which can turn out to be a great focus for the synthesis of anti-tubercular treatment. Inspired from the offering biological actions of phytoconstituents from Allium sativum, the current research concentrates on looking at novel lead compounds from the plant. Molecular docking studies were carried out employing specific phytoconstituents from A.sativum with the protein InhA target. Ajoene shows much more encouragingresults with a Mol Dock rating of 80.6047Kcal/mol, as opposed to the typical initial line drug isoniazid (Moldock score: -58.7028 Kcal/mol). Molecular docking prediction indicate that Ajoene could be formulated into a possible treatment drug for Mycobacterium tuberculosis. <![CDATA[Implications arising from the use of <strong><em>Cymbopogen proximus</em></strong> ; proximal on placenta of pregnant Albino rats]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100330&lng=pt&nrm=iso&tlng=pt ABSTRACT Halfa-bar (Cymbopogon proximus), is an aromatic grass widely growing in Upper Egypt. This herb is recommended for medical purposes as an effective diuretic, renal or abdominal antispasmodic agent. Objectives of this study: Evaluate the potential effects of Halfa-bar on the pregnant albino rats during the gestation period. Material and methods: The virgin female rats mated with male then the pregnant rats treated orally with Human Equivalent Dose (HED) of the proximol which equivalent 0.05 mg/ kg rat from 5th -18th Gestational Day (GD). At day 20 of pregnancy, all rats were anesthetized and killed to obtained maternal -fetal data (placenta). Results: The current study indicated that, there is statistically significant (P ≤ 0.05) reduction in the treated placental weight. Also, the light microscopic examination of the placental specimens using haematoxylin and eosin (H&amp;E) staining revealed the presence of various vacuoles in the cytoplasm and nuclei of the giant cells. There is an increase in the number of apoptotic cells and irregular dilatation of maternal sinusoids in the labyrinth zone. Else, microscopic investigation showed a depletion of the glycogen content in the basal and labyrinth layers and a positive caspase-3 in the spongitrophoblast cells. In the treated group, reduction in level of catalase activity (CAT) and significant elevation (P ≤ 0.05) in the level of malondialdehyde (MDA) were recorded. Conclusion: The pathological effects in placenta may be due to the accumulation of proximal and transplacental passage. <![CDATA[The immunogenicity and anti-tumor effects of a lung cancer DNA vaccine harboring a MUC-1 and GM-CSF fusion gene]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100331&lng=pt&nrm=iso&tlng=pt ABSTRACT DNA vaccines have been shown to be an effective approach to induce antigen-specific cellular and humoral immunity. However, the inability of DNA vaccines to elicit strong immune responses in clinical trials limits the application of DNA vaccines. Here, we developed a new DNA vaccine based on MUC1, which has been suggested as a potential target for lung cancer therapy, and we enhanced the potency of the DNA vaccine by including granulocyte-macrophage colony-stimulating factor (GM-CSF) as an adjuvant. A series of DNA plasmids encoding MUC1, human GM-CSF and their conjugates were constructed and injected into female mice intramuscularly (i.m.). This action was followed by an electric pulse. The humoral and cellular immune responses after immunization were examined by ELISA and ELISPOT, respectively. To evaluate the therapeutic efficacy of the plasmids, a mouse model with a MUC1-expressing tumor was designed. Mice vaccinated with the MUC1-GM-CSF plasmid generated the strongest MUC1-specific humoral and cellular immune responses. Furthermore, these vaccinations inhibited the growth of MUC1-expressing tumors and prolonged mouse survival. These observations emphasize the potential of GM-CSF as an adjuvant for DNA vaccines and of vaccines based on MUC1 and GM-CSF as a promising treatment for lung cancer. <![CDATA[Isolation, Identification and Molecular Characterization of Highly Pathogenic Newcastle Disease Virus From Field Outbreaks]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100332&lng=pt&nrm=iso&tlng=pt ABSTRACT Newcastle disease (ND) is a major infectious disease of the poultry caused by a virulent strain of Avian Paramyxovirus - 1, that is a single strand non-segmented negative sense RNA virus. ND virus is major threat to the poultry industry in many countries of the world. The study was aimed to isolate and identify Newcastle disease virus (NDV) by using a haemagglutination inhibition (HI) test and reverse transcription-polymerase chain reaction (RT-PCR) assay. A total 100 samples of infected and dead birds were collected from different poultry farms. The weight of the birds was ranged 1000-1200g. The birds were divided into 3 groups. Haemagglutination assay (HA) was performed to detect the presence of NDV in suspension of infected homogenized tissues and it was found that HA is not the best method to detect the virus when it is in trace amounts. RT-PCR using NDV specific primers analyzed different clinical and postmortem samples. Reverse transcriptase polymerase chain reaction and specific primers was used for determining the presence of viruses. It was found that the virus was present in most of the infected samples except the serum of infected birds. During multiple sequence alignment (MSA) it was found that, our isolates have high homology (98%) with other reported NDV isolates. Phylogenetic analysis revealed that our isolate was closely related with viscerotropic velogenic types of NDV, which are highly pathogenic Newcastle disease virus. <![CDATA[Effect of Heavy Metals on Liver, Kidney, Gills and Muscles of <strong><em>Cyprinus carpio</em></strong> and <strong><em>Wallago attu</em></strong> inhabited in the Indus]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100334&lng=pt&nrm=iso&tlng=pt ABSTRACT: The present study was aimed to evaluate the effect of heavy metals on an important tissue of two fish species Cyprinus carpio and Wallago attu, sampled from the Indus river, Mianwali District, Pakistan. The concentration of selected heavy metals Fe, Cr, Cu, and in gills, muscles, kidney and liver was compared with an International standard of food fish. The overall metal concentrations among different weight categories in C. carpio were in the order of Fe &gt; Cu &gt; Cr &gt;. In W. attu the overall accumulation of these metals were, in order of Fe &gt; Cu &gt; Cr &gt; Pb The order of accumulation of metals in gills and muscle of C. carpio was Fe &gt; Cr &gt; Pb &gt; Cu; kidney and muscles of W. attu was Fe &gt; Cr &gt; Cu &gt; Pb; liver Fe &gt; Cu &gt; Cr &gt; Pb. An increasing trend of concentration of iron, copper, chromium and lead occurred with an increase in weight of C. carpio and W. attu. There was a significant difference in the accumulation of heavy metals in different organs of both species (p&lt;0.01). All studied heavy metals except Cr were within permissible limits described by various international agencies like WHO, FAO and FEPA in edible tissues of C. carpio and W. attu. <![CDATA[Rat dental pulp stem cells: isolation and phenotypic characterization method aiming bone tissue bioengineering]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100335&lng=pt&nrm=iso&tlng=pt ABSTRACT: Dental pulp stem cells (DPSC) have been showing a considerable potential for regenerative medicine. Pulps were collected from lower incisors (n=2) through direct access of the tooth pulp chamber. The isolated cells were cultured in alfa-MEM 10% FBS, in standard culture conditions. At the third passage, DPSC were characterized by flow cytometry (MHCI, CD54, CD73, CD90, CD45, CD11 and CD34); RT-PCR for Nanog gene; and their differentiation capacity in osteogenic, adipogenic and chondrogenic cell lines. Isolated cells exhibited adhesion capacity to plastic; fusiform morphology, and 80% confluence reached in approximately 3 days. These cells have also revealed positive expression for CD54, CD73 and CD90 markers; and negative expression for CD11, CD34 and CD45. Nanog expression was detected by RT-PCR, expected for a mesenchymal stem cell profile. DPSC chondrogenic differentiation was confirmed by positive staining in Alcian Blue; lipidic droplets stained with oil red confirmed their capacity to differentiate in adipogenic fate; while mineralized beads, stained with alizarin red, confirmed their differentiation in osteogenic phenotype. These results indicate the viability of the isolation and expansion of rat DPSC following this method, and osteogenic differentiation potential opens new perspectives for in vivo studies and the use of these cells in cellular therapies and tissue bioengineering, aiming bone repair. <![CDATA[Influence of mulberry leaf extract on serum adiponectin, visfatin and lipid profile levels in type 2 diabetic rats]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100336&lng=pt&nrm=iso&tlng=pt ABSTRACT The effect of ethanolic mulberry leaf extract (MLE) and mulberry leaf powder (MLP) on glycemic control, serum adiponectin, visfatin and lipid profile in type2 diabetic rats have been investigated. 30 male wistar rats randomly divided into 5 groups. One group was randomly assigned as control (I) and diabetes was induced in others by administration of streptozotocin (STZ) (55 mg/kg body weight) 15 minutes after the administration of nicotinamide (110 mg/kg body weight) intraperitoneally. Finally, fasting blood glucose (FBG), lipid profile, adiponectin and visfatin were assessed after 6 weeks. Lipid profiles including serum FBG, total cholesterol, triglycerides, low-density lipoprotein cholesterol (LDL-c), very low-density lipoprotein cholesterol (VLDL) and visfatin significantly decreased and high-density lipoprotein cholesterol (HDL-c) and adiponectin increased in the two groups of treated diabetic rats in comparison to the diabetic control (p&lt;0.05). For all the investigated factors, there was no significant difference between two treatment methods. However, MLP was more effective than MLE in improving visfatin. Results showed that MLE and MLP possess hypoglycemic and hypolipidemic activities and play an important role in regulating the secretion of adipokines such as adiponectin and visfatin. <![CDATA[Identification of a fat-soluble fraction of <strong><em>Sparganium stoloniferum</em></strong> that inhibits cervical cancer HeLa cells]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100337&lng=pt&nrm=iso&tlng=pt ABSTRACT Context The rizoma of Sparganium stoloniferum has been used as a traditional Chinese medicinal herb for thousands of years. Sparganium stoloniferum is a stasis-breaking drug to treat a wide range of diseases including cancer,however, its activity of extract on cervical cancer HeLa cells and the mechanisms remains unknown. Objective This study aimed to screen Sparganium stoloniferum extract for its inhibitory effects on cervical cancer HeLa cells. Materials and methods Sparganium stoloniferum was extracted with 95% ethanol under reflux, and the extracts were preliminary separated by silica gel column chromatography. MTT assay and flow cytometry were used to determine the inhibitory effects of three fractions on HeLa cells. In addition, GC-MS was performed to analyze the chemical composition of the active fraction. Results Sample II showed a dose-dependent inhibition of HeLa cell growth, with an inhibition rate of more than 30%, whereas the inhibition rates of Samples I and III were less than 30%. Interestingly, Samples I and III had no effect on apoptosis, in contrast to Sample II, which significantly promoted HeLa cell apoptosis, in a dose-dependent manner. GC-MS was performed to analyze the chemical components of Sample II: steroids were found to be the major components with a relative content of 73.905%, while six known compounds were obtained for the first time. Discussion and conclusion This study provided a novel insight for further research of active fractions of Sparganium stoloniferum, in accordance with the basic principle and theory of traditional Chinese medicine (TCM), and will promote the shift of effective substance study, from monomeric chemical compounds to active fractions. <![CDATA[Characterization Of A Novel Hydrolytic Enzyme Producing Thermophilic Bacterium Isolated From The Hot Spring Of Azad Kashmir-Pakistan]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100338&lng=pt&nrm=iso&tlng=pt ABSTRACT A thermophilic bacterium (TP-2) was isolated from the Tatta Pani hot spring in Azad Kashmir and was characterized using phenotypic and genotypic characters. The strain developed cream colored, round, smooth, flat and slimy colonies while the cells were Gram positive rods that ranged in size from about 2.1-3.6 μm to 0.2-0.3 μm in width. Sequence analysis of its 16S rRNA gene showed that isolate TP-2 had 89% homology with Geobacillus debilis. It grew within pH range of 5.5 to 8.5 with optimum growth at pH 7.0. The isolate showed optimum growth at 65ºC and gave positive results for gelatin hydrolysis (GEL), ortho nitrophenyl-β-D-galactopyranosidase (ONPG), and nitrate production and produced acid from sucrose, glucose and maltose. It utilized glucose, fructose, maltose, lactose, sucrose, xylan, starch, filter paper and carboxymethylcellulose as sole carbon source. Isolate TP-2 produced significant amount of industrially important enzymes i.e. extracellular α-amylase, CMCase, FPase, Xylanase, Protease and Lipase and intracellular CMCase and FPase. <![CDATA[The Effect of PM <strong><sub>10</sub></strong> on Ischemia- Reperfusion Induced Arrhythmias in Rats]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100339&lng=pt&nrm=iso&tlng=pt ABSTRACT Epidemiological studies show that particulate matter (PM) is the principal instigator of some adverse clinical symptoms involving cardiovascular diseases. PM exposure can increase experimental infarct size and potentiate myocardial ischemia and arrhythmias in experimental MI models such as ischemia-reperfusion (I/R) injury.The present study was aimed to evaluate the effects of particulate matter (PM10) on ischemia- reperfusion induced arrhythmias with emphasis on the protective role of VA as an antioxidant on them. Male Wistar rats were divided into 8 groups (n=10): Control, VAc, Sham, VA, PM1 (0.5 mg/kg), PM2 (2.5 mg/kg), PM3 group (5 mg/kg), PM3 + VA group. Within 48 hours, PM10 was instilled into trachea in two stages. Then the hearts were isolated, transferred to a Langendorff apparatus, and subjected to global ischemia (30 minutes) followed by reperfusion (60 minutes). The ischemia- reperfusion induced ventricular arrhythmias were assessed according to the Lambeth conventions.In the present study,the number, incidence and duration of arrhythmiasduring30 minutes ischemia were demonstrated to be more than those in the reperfusion stage. PM exposure increased significantly the number, incidence and duration of arrhythmias in the ischemia and reperfusion duration. Vanillic acid reduced significantly the number, incidence and duration of arrhythmias during the ischemia and reperfusion period.In summary, the results of this study demonstrated that the protective and dysrhythmic effects of VA in the PM exposure rats in I/R model are probably related to its antioxidant properties. <![CDATA[Histological Observation and Expression Patterns of antimicrobial peptides during Fungal Infection in <strong><em>Musca domestica</em></strong> (Diptera: Muscidae) Larvae]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100340&lng=pt&nrm=iso&tlng=pt ABSTRACT Housefly, Musca domestica, has a complicated immune system. However, its underlying operating mechanism remains elusive. Candida albicans is a major pathogen affecting humans by causing deep infection fungous disease, but it is non-symbiotic in houseflies. To investigate the C. albicans infection process in housefly, the changes in morphological and histological and expression patterns of antimicrobial peptide were monitored to indicate the insect's response to fungal infection. The results showed that scattered brown spots were comprising melanized encapsulation and encapsulated fungal cells were initially observed at the inner side of larvae's body wall 3 h post-infection (PI). Between 6 and 36 h PI, the whole body of larvae was densely covered with the brown spots, which then gradually disappeared. The majority had disappeared at 48 h PI. Some fungi colonized in the gaps between the body wall and the muscle layer, as well as among muscle fibers of the muscle layer at 12 h PI and hyphal was observed at 18 h PI. These fungi colonized distribution changed from a continuous line to scattered spots at 24 h PI and virtually disappeared at 48 h. The results of quantitative PCR analysis revealed that in coordination with the variation during the infection, the expression levels of four antimicrobial peptides were up-regulated. In conclusion, C. albicans infection in M. domestica larvae involved the following stages: injection, infection, immune response and elimination of the pathogen. The rapid response of antimicrobial peptides, melanized encapsulation and agglutination played a vital role against the pathogenic invasion. <![CDATA[Antioxidant Status, Lipid Peroxidation and Testis-histoarchitecture Induced by Lead Nitrate and Mercury Chloride in Male Rats]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100341&lng=pt&nrm=iso&tlng=pt ABSTRACT This study was done to evaluate the effects of lead nitrate and mercury chloride in testis tissues of Wistar rats. Lead nitrate and mercury chloride are widely used heavy metals in industry. Oral lead and mercury administrations to adult male rats at doses 45 mg/kg bw and 0.02 mg/kg bw, respectively for 4 weeks caused a significant increasing in MDA levels and antioxidant enzyme activities (SOD, CAT, GPx and GST). The MDA levels and acivities of antioxidant enzymes was lower in rats that were administrated by lead nitrate than mercury chloride treated group. Light microscopic analyses revealed that lead nitrate and mercury chloride induced numerous histopathological changes in testis tissues of rats. Histopathological observations of the testis tissues showed that mercury chloride caused more harmful effects than lead nitrate, too. The results indicate that lead nitrate and mercury chloride have reproductive toxicity, in male rats at the tested doses. The effect which we observed applying the lead nitrate and mercury chloride together, was more greater than when we used them alone. <![CDATA[Protective Effects of Edaravone against Methamphetamine-Induced cardiotoxicity]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100342&lng=pt&nrm=iso&tlng=pt ABSTRACT Methamphetamine (METH) is widely abused in worldwide. METH use could damage the dopaminergic system and induce cardiotoxicity via oxidative stress and mitochondrial dysfunction. Edaravone, a sedative-hypnotic agent, is known for it's antioxidant properties. In this study we used edaravone for attenuating of METH-induced cardiotoxicity in rats. The groups (six rats in each group) were as follows: control, METH (5 mg/kg IP) and edaravone (5, 10 and 20 mg/kg, IP) was administered 30 min before METH. After 24 hours, animals were killed, heart tissue was separated and mitochondrial fraction was isolated and oxidative stress markers were measured. Edaravone significantly (p&lt;0.05) protected the heart against lipid peroxidation by inhibition of reactive oxygen species (ROS) formation. Edaravone also significantly (p&lt;0.05) increased the levels of heart glutathione (GSH). METH administration significantly (p&lt;0.05) disrupted mitochondrial function that edaravone pre-treatment significantly (p&lt;0.05) inhibited METH-induced mitochondrial dysfunction. Protein carbonyl level also increased after METH exposure, but was significantly (p&lt;0.05) decreased with edaravone pre-treatment. These results suggested that edaravone is able to inhibition of METH-induced oxidative stress and mitochondrial dysfunction and subsequently METH-induced cardiotoxicity. Therefore, the effectiveness of this antioxidant should be evaluated for the treatment of METH toxicity and cardio degenerative disease. <![CDATA[Ocean Dwelling Actinobacteria as Source of Antitumor Compounds]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100343&lng=pt&nrm=iso&tlng=pt ABSTRACT Extending over decades, research has been of great focus and enormous progress on exploring the ocean for natural products from marine actinobacteria. Attraction towards alternative medicine has led to improvements in natural product discovery. With great potential to survive in extreme environments, marine actinobacteria, efficiently produce an array of metabolites with diverse bioactivity by evolving the secondary metabolic pathways. Exploiting the secondary metabolite producing potential of actinobacteria, many compounds with antitumor, antibacterial, antifungal, antimalarial, antiprotozoal, antiparasitic, antiviral, anti-parasitic, anti-inflammatory activities has been discovered. Efforts in bioprospecting alternative sources of natural products have thus led to several explorations and improvements in technologies which has decreased the bottle neck difficulties in the drug discovery process. Emphasizing on the recent advancements in bioactive compound production in actinobacteria, this paper comprises a review of the available literature, compiles the antitumor compounds from marine actinobacteria with brief discussions and the perspectives to develop better antitumor compounds which would stimulate further research. <![CDATA[The Dynamic of some Biochemical Indicators at Species of Medicinal Plants from Areas of Suceava County, Romania, Emerged under the Mining Pollution Influence]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100344&lng=pt&nrm=iso&tlng=pt ABSTRACT It is known the fact that the mining causes huge damages on the environment having negative biologic effects extremely severe on some groups of organisms of the food chain. The pollutants causes major damages at the level of leaves' cuticles, the vegetation reacting through irreversible modifications at the tissue's level and at substances and energy metabolism, like a riposte to the apparition of the stress induced by the ROS, stress that creates real disturbances on physiological and biochemical processes. Considering the fact that, on the one hand, the activity of oxidoreductases is strongly influenced by the presence of the pollutants factors, with direct effect on the oxidative's stress production, and on the other hand that the plants react at ROS accumulation, through self-protection and adaptability mechanisms, the aim of this study consisted in monitoring the activity of some oxidoreductases in the leaves of plantain and mint species harvested in 2015 from different mining areas of Suceava County. In parallel was determined the dehydrogenase activity of the soil, the samples being collected from the same areas subdued to anthropic pollution, using like reference swatch, samples from Putna. The obtained results signalize the accumulation of some quantities highly arisen by free radicals of oxygen at investigated species, the enzymatic activity being significantly higher in the mining plant comparatively to the control areal. The microbial dehydrogenases from the soil samples were remarked through different activities based on the types of heavy metals that pollute each harvesting area and on the deepness profile. <![CDATA[Optimization of Lipase-Catalyzed Transesterification of Cotton Seed Oil for Biodiesel Production Using Response Surface Methodology]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100400&lng=pt&nrm=iso&tlng=pt The aim of this work was to study the biodiesel production from cotton seed oil by lipase produced by Pichia guilliermondii lipase, which was immobilized onto hydrophobic magnetic particles (HMPs). The optimum reaction conditions were determined for lipase dosage, methanol-to-oil molar ratio, temperature and water content. Using response surface methodology, a quadratic polynomial equation was obtained for fatty acid methyl esters (FAMEs) content by multiple regression analysis. Verification experiments confirmed the validity of the predicted model. The optimal conditions for the enzymatic transesterification were temperature of 38.76℃, 31.3% immobilized lipase, 10.4% water content, and a methanol-to-oil molar ratio of 4.715:1. The gas chromatography- mass spectrometry showed that biodiesel was mainly composed of the methyl esters of hexadecanoic, 9,12-octadecadienoic and 9-octadecadienoic acid. <![CDATA[Cultivation and Biological Characterization of Chicken Primordial Germ Cells]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100401&lng=pt&nrm=iso&tlng=pt The purpose of this work was to investigate the isolation, culture process of chicken gonadal primordial germ cells (PGCs) and study their biological characterization. PGCs were harvested from 5.5-day-old chicken embryonic genital ridges and explanted onto chicken embryonic fibroblasts (CEFs). The results showed that the primary cultivation of chicken PGCs on their own gonadal stroma cells were better than CEFs at first two days for reproduction. The conditioned media supported the growth and colony formation of PGCs for a prolonged time in vitro and maintained a normal diploid karyotype, which were positively stained by alkaline phosphatase (AKP), periodic acid Schiff (PAS) and reacted with anti-SSEA-1, SSEA-3, Oct4, Blimp1 and Sox2. Real-time PCR showed that they expressed the stage specific genes CVH, Blimp1 and Dazl, the stem cell specific genes Sox2, Pouv and Nanog. They also formed the embryoid bodies (EBs). These results suggested that the chicken PGCs cultured in vitro not only had strong self-renewal ability, but also had the potential capability of multi-lineage differentiation. <![CDATA[Multipotent male germline stem cells (mGSCs) from neonate porcine testis]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100402&lng=pt&nrm=iso&tlng=pt ABSTRACT Spermatogonial stem cells (SSCs) are the foundation of spermatogenesis, during which unlimited spermatozoa is produced daily derived from SSCs in the testis throughout life of the male. Germline stem (GS) cells can be isolated from spermatogonia, which shared the characteristics of SSCs and embryonic stem cells (ESCs), and can be passaged stably in vitro. The study of GS cells contributes to understanding spermatogenesis process. However, little is known about the GS cells in domestic animals. Here, we report the successful establishment of a serum- and feeder-free system for multipotent male GS cells (mGSCs) from postnatal porcine testis. These cells expressed pluripotent markers, such as Oct4, Nanog, C-myc, and germline-specific markers including Vasa, CD90, CD49f, Gfrα1, Plzf and Dazl. Then we assayed the developmental potential of these cells in vitro. The porcine multipotent male germline stem cells (pmGSCs) can form embryoid bodies (EBs) by suspension culture. Immunofluorescence analysis showed that the EBs differentiated into neuron-specific enolase (NSE, ectoderm), α-actin (mesoderm), and Pdx1 (endoderm) positive cells. These cells induced by 10-6 M retinoic acid (RA) could be differentiated into spermatid-like cells which were positive for Acrosin. The pmGSCs has been cultured over 14 passages. Thus, we have established a long-term culture system for pmGSCs. This culture system provides a platform for the study of porcine mGSCs. <![CDATA[Micropropagation of Endangered and Decorative Species Dianthus pinifolius Sibth. et Sm.]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100403&lng=pt&nrm=iso&tlng=pt The aim of this study was to establish an efficient protocol for the in vitro propagation of the endangered, endemic and decorative species Dianthus pinifolius Sibth. et Sm. The effects of different concentrations of 6-benzylaminopurine (BAP) and naphtalenacetic acid (NAA), and different explant types (single-node cuttings, terminal buds and shoot cuttings) on shoot multiplication were examined on Murashige and Skoog (MS) and half-strength MS media. The best results were obtained for shoot cuttings on the MS medium supplemented with 0.5 mg/L BAP and 0.1 mg/L NAA, achieving a maximum rate of regeneration (100%) and a total of 15.4 newly-developed shoots per explant. The highest rooting rate (96.7%) was obtained on MS medium containing 1 mg/L NAA, while the acclimatization of the microplants obtained to ex vitro conditions was successful (88.9%). <![CDATA[Identification of Suitable Reference Genes for Gene Expression Normalization in <em>Jatropha curcas L</em> During Development and Under Stress Conditions Using Real Time Quantitative PCR]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100404&lng=pt&nrm=iso&tlng=pt Jatropha curcas L represent a potential source of raw material for the production of biodiesel. The aim this study was to find potential candidate reference genes in J. curcas tissues. Three softwares were utilized to verify which would be the most stable reference genes in qPCR assay: GeNorm, NormFinder and BestKeeper. The most stable reference genes in developing J. curcas seeds suggested by GeNorm were GAPDH, UCP, actin. However, the best combinations of stable genes in each tissue were identified separately under stress conditions: EF1-α, PP2A2 and GAPDH in total stress, however, in SA stress, four genes were required for normalization: PP2A2, EF1-α, GAPDH and PUB. In PEG stress, four genes also were required: PP2A2, EF1-α, GAPDH and PUB, while in NaCl stress, five genes were necessary: PP2A2, GAPDH, EF1-α, PUB and Tβ2. These results are in accordance with two other programs used in this study (NormFinder, BestKeeper). In addition, the transcript levels of Jc-SRG-2 seem to be more correlated with stress responses than changes in transcript levels of Jc-SRG-1, mainly of leaves in exposure to 3-12 h on PEG and NaCl stress. Taken together, GAPDH and PP2A2 were regarded as being the best reference to provide guidelines for the selection of potential references genes under these study conditions. <![CDATA[Comparisons Between Different Methods in Measuring Enzyme Similarity for Metabolic Network Alignment]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100405&lng=pt&nrm=iso&tlng=pt Metabolic network alignments enable comparison of the similarities and differences between pathways in two metabolic networks and help to uncover the conserved sub-blocks therein. Such analysis is important in the understanding of metabolic networks and species evolution. The fundamental parts of metabolic network alignment algorithms all involve comparisons of the similarity between two enzymes as a similarity measure of network nodes. As a result, the study of methods for measuring enzyme similarity becomes highly relevant. Currently, two approaches are mainly used to measure enzyme similarity. One of the methods is based on similarity measures of gene or protein sequences; the other is based on enzyme classification. In this study, multiple metabolic network alignments were performed using both the methods. The results showed that, in general, the sequence similarity method yielded higher accuracy, especially with respect to reflecting evolutionary distances. <![CDATA[Comparative Microbicidal Efficacy of Fractionated Extracts from In Vitro and In Vivo Raised Cells of Tinosporacordifolia Against MDR Pathogens]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100406&lng=pt&nrm=iso&tlng=pt The present study was conducted to explore the hidden potential of natural products synthesized in the medicinal plant Tinosporacordifolia. This plantis prioritized by National Medicinal Plant Board, New Delhi. Leaf and inter nodal segments were inoculated on MS Medium fortified with IBA (1.0 mg/L) produced callus after four weeks. The calli were brown due to phenolic substance secreted by the explant. This problem was overcome by using adjuvant PVP (0.1%). Further, secondary metabolites were isolated from callus and field leaf through soxhlet extractor and fractionated by using column chromatography. The antibacterial activity of these fractioned extracts from Tinosporacordifolia callus and leaf were seen against multi drug resistance bacteria viz., Escherichia coli (ATCC 25922), Pseudomonas aeruginosa, (ATCC 27853) &amp; Staphylococcus aureus (ATCC 29213) and against plant pathogenic fungus Fuseriumoxisporum(MTCC 8608) and Sclerotiniasclerotiorum (MTCC 8785). All fractionated extracts showed antimicrobial activity but callus extracts were proved to be best in compare to leaf extracts. Furthermore, we are trying to analyze different bio active compounds through GCMS. <![CDATA[Production and Characterization of Bacteriocin Produced by Lactobacillus Viridescence(NICM 2167)]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100407&lng=pt&nrm=iso&tlng=pt The present study focused on the production optimization of bacteriocin by Lactobacillus viridescence NICM 2167 followed by its purification and characterization. The bacteriocins are antimicrobial peptides produced by many Gram positive and Gram negative bacteria.The bacteriocin produced by LAB (lactic acid bacteria) received attention in recent years due to their potential application as natural preservatives in food. Bacteriocinproduced by Lactobacillus viridescence showed broad range of antimicrobial activity against food borne pathogens. Production parameters were optimized showing highest production of bacteriocinin MRS broth with pH= 7.0 incubated at 37°C for 48 h. Bacteriocin was purified in two steps involving ammonium sulphate precipitation followed by gel filtration using Sephadex G-100. Purified bacteriocin with single band on SDS-PAGE showed molecular weight of 8.3 kDa. This purified bacteriocin was stable over wide range of pH (4-10) as well as temperatures (4°C-121°C) suggesting it as a potent candidate for preservation of various foods. <![CDATA[Down-Regulation of NFkB, Bax,TGF-β, Smad-2mRNA expression in the Livers of Carbon Tetrachloride Treated Rats using Different Natural Antioxidants]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100408&lng=pt&nrm=iso&tlng=pt The objective of this study is to examine whether silymarin alone or in combination with chlorogenic acid and/ or melatonin plays a modulatory role against apoptotic damage in rats liver induced by of CCl4. The present work revealed that CCl4 induced elevation of in Bax, Smad, TGF-β and NFkBhepatic mRNA expression, administration of silymarin alone down regulates these expressions. Treatment with chlorogenic acid and/ or melatonin along with silymarin produced best results in this concern. Bcl-2 expression was down regulated by CCl4 whereas concurrent treatment of chlorogenic acid and/ or melatonin along with silymarin increased this expression. On conclusion, the use of chlorogenic acid and/ or melatonin potentiates the anti-apoptotic action of silymarin. <![CDATA[Effect of Gamma Radiation on Growth and Metabolic Activities of <em>Arthrospira platensis</em>]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100409&lng=pt&nrm=iso&tlng=pt This work aimed to study the influence of gamma radiation on the growth and production of some active substances of Arthrospira platensis. Biomass production was significantly inhibited (p ≤ 0.05) by 21 and 34%, with respect to the control at 2.0 and 2.5 kGy, respectively. Chlorophyll-a content showed 11% reduction at 2.5 kGy compared to the control. As a result of growth and Chl-a inhibition, chlorophyll productivity recorded a continuous significant decrease below the control in the cells exposed to 1, 1.5, 2 and 2.5 kGy by 8, 12, 15 and 25%, respectively after 15 days of incubation. In addition, phycobillins productivity showed significant decrease by 10 and 36% below the control at 2 and 2.5 kGy of gamma radiation, respectively. Protein production decreased significantly by 24% at 1.5 kGy; low doses of gamma irradiation (0.5, 1.0 and 1.5 kGy) induced carbohydrate production by 106, 246 and 146%, respectively. Lipid content increased significantly over the control at 0.5 kGy of gamma irradiation by 22%, which was decreased at higher doses. Interestingly, carotenoid productivity showed significant increase at all used gamma doses up to 155% over the control. <![CDATA[Enhanced Accumulation of Scopoletin in Cell Suspension Culture of Spilanthes acmella Murr. Using Precursor Feeding]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100410&lng=pt&nrm=iso&tlng=pt In this study, the various concentrations of casein hydrolysate (25, 50, 75, 100 mg/L) and L-phenylalanine (50, 100, 150, 200 µM/l) were incorporated in MS containing 15 µM BA plus 5 µM 2,4-D for enhancement of secondary metabolites in cell culture of Spilanthes acmella. The presence of casein hydrolysate in the nutrient medium improved the growth of cell biomass and the production of scopoletin. The addition of casein hydrolysate up to 75 mg/L stimulated the accumulation of scopoletin, but increasing excess 75 mg/L the level of casein hydrolysate reduced the production of scopoletin. The addition of L-phenylalanine in the nutrient medium was found to be more effective for production of secondary metabolite in S. acmella. The addition of 50 µM/L of L-phenylalanine in the medium increased scopoletin content to 27.12 ± 0.58 µg/g dry weight, compared to the scopoletin content of control at 7.89 ± 0.61 µg/g dry weight. The highest accumulation of scopoletin was observed in the 100 µM/L L-phenylalanine in cell suspension, which was 4.51 times more than the control. As a result, using moderate concentration of L-phenylalanine was ideal for the production of scopoletin. In general, casein hydrolysate was more effective than L-phenylalanine for production of scopoletin and growth of cell biomass in the cell culture of S. acmella. <![CDATA[Variation of Hydroxyapatite Content in Soft Gelatin Affects Mesenchymal Stem Cell Differentiation]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100411&lng=pt&nrm=iso&tlng=pt Gelatin is a common material used in tissue engineering and hydroxyapatite (HA) has a composition and structure similar to natural bone mineral. HA is also used to increase the adhesion ability of scaffolds. The physical and mechanical properties of gelatin, together with the chemical properties of HA, can affect cell differentiation. The main purpose of this study is to investigate the gene expression of human mesenchymal stem cells (HMSCs) upon culturing on gelatin composite with HA. Low amounts of HA were introduced into the gelatin in order to modulate properties of gelatin. Three types of hydrogel were fabricated by glutaraldehyde crosslinking before lyophilization to produce the porous 3D structure: (1) pure gelatin, (2) 0.5 mg/ml HA in gelatin, and (3) 1 mg/ml HA in gelatin. The fabricated hydrogels were used as scaffolds to cultivate HMSCs for two periods - 24 hours and 3 weeks. The results showed that all types of fabricated hydrogels could be used to cultivate HMSCs. Changes of gene expressions indicated that the HMSCs cultured on the 1 mg/ml HA in gelatin showed neuronal lineage-specific differentiation. <![CDATA[Recurrent Somatic Embryogenesis and Plantlet Regeneration in Psidium guajava L.]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100412&lng=pt&nrm=iso&tlng=pt A simple and efficient protocol for recurrent somatic embryogenesis and plant regeneration is one of the prerequisites for genetic improvement of guava. An efficient reproducible regeneration somatic embryogenesis protocol was developed in four genotypes of Psidium guajava L. using immature zygotic embryo as starter explant. Somatic embryogenesis induction was obtained on MS basal medium supplemented with 2.0 mgL-1 2, 4-D, 400 mgL-1 L-glutamine, 6% sucrose and 500 mgL-1 Malt extract. Following SE induction different developmental stages of somatic embryos (Globular, heart-shaped, torpedo, cotyledonary) was directly obtained and further recurrent embryogenesis also obtained upon prolonged incubation in induction media. Addition of polyethylene glycol (50 mgL-1) significantly improved the embryos maturation in MS supplemented with and 3% sucrose. The regeneration in MS medium supplemented with BAP (0.5 mgL-1), NAA (0.2 mgL-1), casein hydrolysate (100 mgL-1) and 3% sucrose. High plant regeneration frequency and intensity of somatic embryos (58.5%) obtained. Plant maturation on MS medium supplemented with BAP 2.0 mgL-1 with 2% sucrose. The rooted plants was successfully acclimatize in the greenhouse with a survival rate of 85%. This somatic embryogenesis protocol developed would be helpful in establishment of genetically modified guava aimed at seedlessness, increased shelf life and wilt disease. <![CDATA[Biological properties of Endophytic Fungi]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100413&lng=pt&nrm=iso&tlng=pt The endophytic fungus is have rich source of secondary metabolites which act as biological active agent in the higher plants. The fungal derivatives play vital part in human life and their compounds are the source of drug for cancer, microbial and viral diseases. The natural compounds from the endophytes acts as growth inhibitor of plant pathogenic organism. Endophytes are rich sources of natural products which are used in agriculture (plant growth and insecticidal), pharmaceutical industries and also used for phytoremediation. <![CDATA[Morphological and Biochemical Answer of the Wheat Seeds at Treatment with 2,4-Dinitrophenol and Potassium Iodate]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100414&lng=pt&nrm=iso&tlng=pt Oxidative stress can be regarded as an imbalance between the amount of reactive oxygen species (ROS) and the ability of a biological system to eliminate the toxic species and repair the resulting damages. Since the germinating seeds and the resulted seedlings are rich in enzymes, whereas the treatment with chemicals affects much the seed germination, producing also ROS, we evaluate here the influence of 2,4-dinitrophenol (DNP) and potassium iodate (KIO3) on wheat germination (Triticum aestivum L.) and seedlings growth. Germination rate, the masses and heights of the 7 day old seedlings, as well as the activity of some enzymes involved in the oxidative stress such as peroxidase, catalase and superoxide dismutase were measured seven days after the chemical treatment. The treatment of the wheat seeds with 10-5 - 10-3 M solutions of DNP resulted in a relative concentration-dependent inhibition of the germination, with a concomitant stimulation of the weight and height of viable seedlings. The Gasparom variety treated with 10-5 M KIO3 showed a slight increase in the germination rate in comparison with the control batch. The two tested substances determined a significantly modified response of the oxidative stress enzymes, especially in the seeds treated with 10-4 and 10-3 M solutions. <![CDATA[The correlation between the central carbon metabolic flux distribution and the number of shared enzyme regulators in Saccharomyces cerevisiae]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100415&lng=pt&nrm=iso&tlng=pt The central carbon metabolic system is the upstream energy source for microbial fermentation. In addition, it is a master switch for increasing the production of metabolites and an important part of the microbial metabolic network. Investigation into the relationship between genes, environmental factors, and metabolic networks is a main focus of systems biology, which significantly impacts research in biochemistry, metabolic engineering, and synthetic biology. To this end, the central carbon metabolic flux under a variety of growth conditions or using strains with various genetic modifications was previously measured in Saccharomyces cerevisiae using 13C tracer technology. However, the measured values were not integrated and investigated further. In this study, we collected and analyzed the metabolic flux rates of the central carbon metabolic system in S. cerevisiae measured in recent studies. We carried out preliminary analyses of flux values of each pathway, performed regression analyses on relationship between different fluxes, and extracted principal component factors of the flux variables. Based on the results, the general characteristics of pathway flux distribution were clustered and explored, and the effects of environmental and genetic factors on the flux distribution were analyzed. Furthermore, this study explored the relationship between similarity in the enzyme's transcriptional regulation and the correlations in the enzyme's reaction flux. Our results provide a foundation for further studies on the control of the central carbon metabolic flux and facilitate the search for targets in metabolic engineering research. <![CDATA[Application of Response Surface Methodology for Optimizing Process Parameters in the Production of Amylase by Aspergillus flavusNSH9 under Solid State Fermentation]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100416&lng=pt&nrm=iso&tlng=pt Amylase is recognized as one of the important commercial enzymes. This group of enzymes has the ability in hydrolyzing starch into smaller oligosacharides. The present work aimed to determine the optimum fermentation conditions for maximum production of crude amylase enzyme by Aspergillus flavus NSH9 employing response surface methodology (RSM).Central composite design (CCD) was applied to determine the optimal fermentation condition with respect to the four main process parameters such as temperature, initial moisture content, pH and the incubation period. Solid state fermentation (SSF) was performed using 5.0 g of sago hampas inoculated with 1x107sporesmL-1following the experimental design obtained using CCD and further optimized by RSM. The initial moisture, pH and temperature showed significant effect on the amylase production (p&lt;0.05). The maximum amylase activity produced was achieved and recorded as 1.055 ± 0.03U mL-1after four days of fermentation period with 100% (v/v) moisture holding capacity, pH 6.5 and temperature at 28°C. The optimum fermentation conditions for amylase production was determined with A. flavusNSH9 on sago hampas. <![CDATA[Biodiversity of Antimicrobial-Producing Actinomycetes Strains Isolated from Dry Dipterocarp Forest Soil in Northeast Thailand]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100417&lng=pt&nrm=iso&tlng=pt The increasing use of antibiotics has led to the development of drug-resistant microorganisms. The emergence of resistant strains has compromised the treatment and control of infectious diseases. As a result, the search and development of a novel class of antibiotic drugs are required. Actinomycetes have been recognized as a richest source of secondary metabolites including antibiotics. The aim of this study was to investigate the diversity of antibiotic-producing soil Actinomycetes from dry dipterocarp forest in Northeast of Thailand. The soil from this area has been known for its poor in nutrients and highly acidic with pH values around 4.5. In such a harsh condition, soil-inhabiting Actinomycetes elevate their protective mechanisms by inducing the production of antimicrobials and other defense compounds. Therefore, dry dipterocarp forest could be served as a potential source for the screening of the novel antimicrobial drugs. Twelve antibiotic-producing strains isolated from soil in Suranaree University of Technology produce antimicrobial agents which are active against wide range of test pathogens including methicillin-resistant Staphylococcus aureus (MRSA). Based on 16S rRNA gene analysis, these strains are close affiliated with the genus Streptomyces (11 isolates) and Nonomuraea (1 isolate). Most of soil isolates show narrow antimicrobial spectrum activity against Gram-positive bacteria. Two isolates, PJ36 and PJ95, exhibit broad antimicrobial spectrum against Gram-positive bacteria, Gram-negative bacteria and yeasts. Phylogenetic tree analysis of 16S rDNA reveals that isolates PJ33, PJ36, PJ43, PJ51, PJ75, PJ76, PJ77, PJ85, PJ88 and PJ95 strains are not cluster with others strain of Streptomyces. They represent a distinct phyletic line which might be suggested the novel strains. This study was the first attempted to isolate antibiotic-producing Actinomycetes from dry dipterocarp forest soil in Northeast Thailand. Most of soil isolates (8 strains) obtained from this study were active against methicillin-resistant Staphylococcus aureus (MRSA). These isolates could be used for the development of new drugs to combat antibiotic resistances. <![CDATA[Influence of Benzyladenine and Thidiazuron on Shoot Regeneration from Leaf and Shoot tip Explants of Sedum sarmentosum Bunge]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100418&lng=pt&nrm=iso&tlng=pt Effects of N6-benzyladenine (BA) or thidiazuron (TDZ) on adventitious shoot regeneration and axillary shoot multiplication of Sedum sarmentosum was investigated. Leaf and shoot tip explants obtained from in vitro-grown shoots of S. sarmentosum were cultured on Murashige and Skoog (MS) medium supplemented with 0, 2.0, 4.0 or 8.0 µM BA or TDZ. Of the two cytokinins studied, BA was found to be more responsive as compared to TDZ with respect to number of shoots produced per explant. High frequency of shoot regeneration (92.2%) with a mean of 12.3 shoots was produced when the leaf explants were cultured on MS medium supplemented with 8.0 µM BA. The highest number of shoots (25.4) was obtained when shoot tip explants were cultured on MS medium devoid of cytokinins after 35 days of culture. For root induction, regenerated shoots were cultured on MS medium supplemented with 0, 2.0, 4.0 or 8.0 µM indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) or α-naphthaleneacetic acid (NAA). The highest number of (17.6) roots per shoot was obtained on MS medium supplemented with 2.0 µM IBA after 30 days of culture. Regenerated plantlets were successfully acclimatized in the greenhouse with 100% survival rate. <![CDATA[Selection of Xilose-Fermenting Yeast Strains]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100419&lng=pt&nrm=iso&tlng=pt ABSTRACT In Brazil, ethanol is obtained by fermentat of sugar cane juice using Saccharomyces cerevisiae. The cane juice extraction generates the bagasse that has been used for obtaining generation biofuel. However, the sugarcane bagasse has 30% pentose that cannot be fermented to ethanol by S. cerevisiae. Thus the aim of this study was to isolate a yeast able to ferment xylose to ethanol. Samples of cane juice and flowers were used for the isolation of 165 strains that were then screened for ethanol production using plate testing. Among them, the ethanol positive strains Wickerhamomyces anomalus, Schizosaccharomyces pombe and Starmerella meliponinorum were selected for a xylose fermentation assay, using a semi-synthetic and bagasse hydrolysate as must. S. meliponinorum and S. pombe produced 0.63 and 2.7 gL-1 of ethanol, respectively, from xylose in a semisynthetic medium. In the medium consisting of bagasse hydrolysate must, 0.67 and 1.1 gL-1 of ethanol were obtained from S. meliponinorum and S. pombe, respectively. All the yeasts produced xylitol from xylose in the semisynthetic medium and S. meliponinorum was that which produced the highest quantity (14.5 g L-1). <![CDATA[Rhamnolipid Biosurfactants Produced by <strong> <em>Pseudomonas Species</em></strong>]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100420&lng=pt&nrm=iso&tlng=pt ABSTRACT: Surfactants are chemical products widely used in our daily life in toothpaste and other personal hygiene and cosmetic products, and in several industries. Biosurfactants are surfactants of biological origin that can be produced by microorganisms and have many advantages, such as low toxicity and high biodegradability, compared to synthetic counterparts. Unfortunately, high production costs limit the use of biosurfactants. Low-cost production is the most important factor for biosurfactants to be able to compete in the global market place. This review presents general information on rhamnolipid biosurfactant produced by Pseudomonas species, as well as on their production and applications. In addition, industrial products and their wastes used for rhamnolipid production are reviewed in detail based on recent studies. <![CDATA[Characterization of the Relationship of CDKL5 with MeCP2 and Dnmt1 in PrimaryRat Cortical Neurons]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100421&lng=pt&nrm=iso&tlng=pt ABSTRACT Cyclin-dependent kinase-like 5 (CDKL5) is a protein kinase that is homologous to mitogen-activated protein kinases (MAPKs) and cyclin-dependent kinases (CDKs). Mutations in the CDKL5 gene cause X-linked infantile spasms and phenotypes that overlap with that of Rett syndrome, which is a neurodevelopmental disorder caused primarily by mutations in the methyl CpG binding protein 2 gene (MECP2). Previous studies in transfected cell lines showed that CDKL5 interacts with MeCP2 and DNA (cytosine-5)-methyltransferase 1 (Dnmt1). However, little is known about the relationships of CDKL5 with interacting proteins in primary neuronal cultures. In this study, we investigated the expression patterns of CDKL5, MeCP2 and Dnmt1, and their interaction in cultured rat cortical neurons. Using real-time PCR analysis, we found that CDKL5, MeCP2 and Dnmt1 have similar expression patterns at the mRNA level. In contrast, the expression patterns of those proteins at the protein level are different and could be inversely correlated, as shown by western blotting. Using co-immunoprecipitation, we further demonstrated that CDKL5 interacts with MeCP2 and Dnmt1 in primary rat cortical neurons. These data suggest that a functional link exists among CDKL5, MeCP2 and Dnmt1 during neuronal development and may provide further insight into the pathogenesis of Rett syndrome. <![CDATA[Crucial Residues Modulating Interface of hBcl-B - hBaxBH3 Heterodimer as Probed by Computational Methods]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100422&lng=pt&nrm=iso&tlng=pt ABSTRACT Cancerous cells develop resistance to cell death by over expression of anti-apoptotic proteins, which are specific to interact with pro-apoptotic and BH3-only proteins of Bcl-2 family. Delineating crucial residues mediating the heterodimer complexes (anti-apoptotic proteins - pro-apoptotic/BH3-only proteins) is indispensable to develop specific antagonists to anti-apoptotic proteins. In these backgrounds, we have herein reported crucial residues of hBaxBH3 and hBcl-B (an anti-apoptotic protein specifically interacts with human Bax but does not interact with human Bak) for hetero dimerization of the polypeptides and as well validated the structural determinants of the polypeptides through variety of virtual 'alanine mutants' and 'switch mutants' by using an array of computational methods. Residues such as D53, S60, E61, K64, E69 and D71 of hBaxBH3 and R45, H50, F53, F54, Y57, M71, S74, V75, R86, V88, T89, F93 and F159 of hBcl-B were found to be crucial residues of the polypeptides for intermolecular interaction leading hetero dimerization. Moreover, 'pharmacophoric residues' for the hBaxBH3 and hBcl-B have also been figured out and rationalized. <![CDATA[Nematicidal activity of <strong><em>Paecilomyces marquandii</em></strong> proteases on infective larvae of <strong><em>Ancylostoma</em></strong> spp]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100423&lng=pt&nrm=iso&tlng=pt ABSTRACT The present study aimed to evaluate the action of Paecilomyces marquandii proteases on Ancylostoma spp L3. White halos in the zymogram confirmed the proteolytic action. Difference (p &lt;0.01) between the number of L3 in the differents groups was found, with 41.4% of reduction of Ancylostoma spp. L3 before 24 hours. <![CDATA[Genetic Variability of <strong><em>Brycon hilarii</em></strong> in a Repopulation Program]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100424&lng=pt&nrm=iso&tlng=pt ABSTRACT Brycon hilarii, popularly called piraputanga in Brazil, is a species distributed throughout the whole basin of the river Paraguay. In recent years, the species has been on a repopulation program due to its remarkable decline as a wild species in the region. Assessment of the genetic diversity of broodstock and fingerling stocks in repopulation programs is basic to avoid genetic impacts on wild populations. The genetic variability of the wild population and of the broodstock and fingerling stocks of B. hilarii in a repopulation program in the river Itiquira MT Brazil will be determined. Seven microsatellite loci produced 52 polymorphic alleles and heterozygosity revealed rates between 0.5794 and 0.7204. FIS did not register any endogamy in the broodstock but it was present in fingerlings and wild populations. Intra- and inter-specific genetic variability rates were higher within each combination but not between groups. Grouping in fingerling groups had a lower density when compared to the others. There is a higher genetic proximity between the natural population and broodstock (0.0237) when the distance between populations was analyzed, even though the two were greatly distant from the fingerling group (0.2622 - 0.2617). Results show that the wild population and the broodstock had high genetic variability and low genetic divergence; contrastingly, fingerlings showed mild genetic variability and great divergence when compared to other groups, indicating that they were not adequately constituted. <![CDATA[A Histological Study on Venom Apparatus of <strong><em>Montivipera xanthina</em></strong> (Gray, 1849) (Serpentes, Viperidae)]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100425&lng=pt&nrm=iso&tlng=pt ABSTRACT This is the first report describing morphological and histological structure of venom apparatus in Montivipera xanthina. The venom gland apparatus of M. xanthina was composed of four distinct parts, main venom gland, primary duct, accessory gland and secondary duct. The main venom gland was composed of acini. The accessory gland was formed by long and ramified tubules. Nine fangs were detected in one half of the upper jaw each at different stage of development. Only one tooth was functional and the eight remaining fangs form a replacement series. The venom apparatus of viperid snakes are the most effective venom-delivery system among vertebrates and have been the focus of scientific interests for many years. Despite this interest, there are few studies including venom-delivery system of venomous snakes. The results presented here may be common among venomous snakes and model of the viperid design of the venom apparatus. <![CDATA[Novel Microsatellite Markers for Brazilian Mangrove Oysters ( <strong><em>Crassostrea gasar</em></strong> ) and their Cross-Amplification in <strong><em>Crassostrea rhizophorae</em></strong>]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100426&lng=pt&nrm=iso&tlng=pt ABSTRACT A microsatellite CT/GT enriched genomic library was developed for Crassostrea gasar and twelve new polymorphic loci were isolated and characterized. The markers were successfully amplified from 25 individuals of Crassostrea gasar and 11 cross-amplified individuals of Crassostrea rhizophorae. There was no evidence of linkage between loci in either species. <![CDATA[Evaluation of Two Fitting Methods Applied for Thin-Layer Drying of Cape Gooseberry Fruits]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100427&lng=pt&nrm=iso&tlng=pt ABSTRACT Drying data of cape gooseberry was used to compare two fitting methods: namely 2-step and 1-step methods. Literature data was also used to confirm the results. To demonstrate the applicability of these methods, two primary models (Page, Two-term-exponential) were selected. Linear equation was used as secondary model. As well-known from the previous modelling studies on drying, 2-step method required at least two regressions: One is primary model and one is secondary (if you have only one environmental condition such as temperature). On the other hand, one regression was enough for 1-step method. Although previous studies on kinetic modelling of drying of foods were based on 2-step method, this study indicated that 1-step method may also be a good alternative with some advantages such as drawing an informative figure and reducing time of calculations. <![CDATA[Spray Drying of Pequi Pulp: Process Performance and Physicochemical and Nutritional Properties of the Powdered Pulp]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100500&lng=pt&nrm=iso&tlng=pt The objective of this work was to optimize the spray drying of pequi pulp using maltodextrin as carrier agent and Tween 80 as surfactant agent. A central composite rotatable design was used to evaluate the influence of inlet air temperature (140 to 200°C), maltodextrin (15 to 30%) and surfactant (0 to 5%) concentration on the process performance and physicochemical and nutritional properties of the dried powdered pulp. The dependent variables were process yield (27.4 - 51.7%), outlet air temperature (106.5 - 135°C), energetic efficiency (29.9 - 44.8%), moisture content (0.25 - 1.43%), water activity (0.09 to 0.21), hygroscopicity (9.1 - 12.1 g adsorbed moisture/100g dry matter), vitamin C content (129.8 - 303.0 mg/g solids pequi) and total carotenoids content (8.2 - 94.9 mg carotenoids/g solids pequi). The spray drying of pequi pulp was optimized for maximum vitamin C and total carotenoids content using response surface methodology, which were attained at 152°C, surfactant concentration of 1% and maltodextrin concentration of 18%. The characterization of the pequi pulp powder obtained at the optimized condition evaluating the particles sizes, bulk density and porosity. The morphology showed spherical and smooth particles with several sizes. <![CDATA[Optimization Medium Composition for Vitamin K<sub>2</sub> by Flavobacterium sp. using Response Surface Methodology and Addition of<em>Arachis hypogaea</em>]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100501&lng=pt&nrm=iso&tlng=pt ABSTRACT The purpose of this research was to enhance the production of vitamin K2 by fermentation optimization and Arachis hypogaea supplementation in Flavobacterium sp. mutant SP-L-01. Optimized culture condition were as follows: 6-days shake-flask culture at 37oC with initial pH value 7.0 ± 0.2, shaking speed in 120 r/min and medium volume of 30 mL with 2% inoculums. After optimization of fermentation medium by response surface methodology (RSM), optimized medium were maltose 23.8 g/l, glucose 9.69 g/l, beef extract 15 g/l, K2HPO4 4.5 g/l,NaCl 3.0 g/l and MgSO4·7H2O 0.3 g/l. Production of vitamin K2 after optimization reached to 10.97 mg/l, which is 79.25% higher than that before optimization (6.12 mg/l). 3 mg/mL of arachis hypogaea was added into the medium at 72 h of shake-flake cultivation, which improved the production of menaquinone-4 (MK4) up to 371% and menaquinone-6 (MK6) up to 149% higher than those of the original medium. D-(+)-catechin, one of the components of arachis hypogaea, was added alone into the medium, which also improved the vitamin K2 synthesis. <![CDATA[Comparative study of a new alkaline L-methioninase production by Aspergillus ustus AUMC 10151 in submerged and solid-state fermentation]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100502&lng=pt&nrm=iso&tlng=pt Twenty four fungal species were screened for their ability to produce alkaline L-methioninase on methionine-glucose liquid medium. Aspergillus ustus AUMC 10151 displayed the highest yield of enzyme (10.8 U/mg protein), followed by A. ochraceus and Fusarium proliferatum. Upon optimization of the submerged fermentation (SmF)conditions, the maximum enzyme yield (18.23 U/mg protein) was obtained on a medium containing L-methionine (0.5%), sucrose (0.95%), KH2PO4 (0.1%) and 175 rpm. Seven agro-industrial by-products were screened as substrates for L-methioninase production under solid-state fermentation (SSF). Wheat bran resulted 38.1 U/mg protein, followed by rice bran (27.6 U/mg protein) and soya bean meal (26.6 U/mg protein). Maximum alkaline L-methioninase (99.56U/mg protein) was achieved at initial moisture content of 71.5%, inoculum size of 2.0 mL of spore suspension, initial pH 8.5, incubation period eight days at 30°C and supplementation of the salt basal medium with pyridoxine(100 μg/mL) and beet molasses (20% v/v). The productivity of L-methioninase by A. ustus under SSF was higher than that of SmF about 5.45 fold under optimum conditions. <![CDATA[Diets Rich in Polyunsaturated Fatty Acids With Different Omega-6/Omega-3 Ratio Decrease Liver Content of Saturated Fatty Acids Across Generations of Wistar Rats]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100503&lng=pt&nrm=iso&tlng=pt Our study evaluated how the consumption of diets with low (LOW group - 0.4/1) or high (CON group - 13.6/1) omega-6/omega-3 ratio across generations (F1 and F2) can modulate liver fatty acid (FA) profile and blood biomarkers. Liver content of α-linolenic acid was higher in animals always fed with LOW diet than animals that changed from CON to LOW diet, which by your time was higher than animals always fed with CON diet. Liver saturated FA concentration decreased in both groups from F1 to F2. In conclusion, both diets were efficient in decreasing the saturated FA liver content across generations, the LOW ratio diet was more effective in reducing blood triglycerides and non-esterified fatty acids, and there was a multigenerational effect of the LOW ratio diet, improving the FA profile even when the offspring start receiving the CON diet. <![CDATA[Effects of Yucca Extract on Feed Efficiency, Immune and Antioxidative Functions in Broilers]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100504&lng=pt&nrm=iso&tlng=pt Effects of yucca extract (YE) on feed efficiency, immune and antioxidative function in Arbor Acres broilers were studied. One hundred and twenty-eight fourteen-day-old broiler chickens were randomly divided into four treatments with four replicates of 8 birds each. These four diets were formulated by adding 0, 100, 200 and 300 mg/kg YE to the basal diet. The results showed that: diets supplemented with 100 and 200 mg/kg YE increased average body weight gain, feed efficiency, IgG, IgM, T-AOC, CAT and SOD levels, and have positive effects on inducing immune organs' maturation. In addition, 100 mg treatment mainly improved the feed efficiency whereas 200 mg treatment mainly acted on immunity and anti-oxidation. In conclusion, YE can be used as a feed additive due to its capability to improve feed efficiency, immune and antioxidative function in broilers. <![CDATA[Development of an L-Lysine Enriched Bran for Animal Nutrition via Submerged Fermentation by Corynebacterium glutamicum using Agroindustrial Substrates]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100505&lng=pt&nrm=iso&tlng=pt L-Lysine is an essential aminoacid added as supplement for animal feed. The aim of this work was to produce an L-Lysine enriched bran using Brazilian agroindustrial byproducts. Both the raw material costs and purification steps were minimized. Firstly, medium composition for the growth of Corynebacterium glutamicum ATCC 21799 was optimized targeting enhanced L-Lysine production - salt, vitamins and nitrogen sources concentrations were tested and selected. Next, UV mutant strains were generated and the best producers were used in formulated media using sugarcane molasses. It was reached a production of 9.3 g/L of L-Lysine with the optimized formulated media. This L-Lyisne rich broth was then impregnated and cyclically reimpregnated in pre-treated solid matrixes (sugarcane bagasse, citrus pulp, brewer spent grain, soybean husk and wheat bran). After processing, it was generated enriched brans with significant amounts of L-Lysine (13.8%, 7.0%, 8.9%, 5.9% and 8.4%, respectively), which has an interesting market potential for animal feed. <![CDATA[<sup>40</sup>K, <sup>226</sup>Ra AND <sup>228</sup>Ra SERIES IN BOVINE AND POULTRY FEED AND IN DICALCIUM PHOSPHATE (DCP) SAMPLES BY GAMMA-RAY SPECTROMETRY]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100506&lng=pt&nrm=iso&tlng=pt The aim of this study is to determine the amounts of radionuclides' activity present in samples of feed for young and adult chicken and feed for dairy and beef cattle, as well as in dicalcium phosphate (DCP) (CaHPO4), used as a nutritional supplement for animal feed. Radioactivity in these samples is due to the presence of the 238U and 232Th radioactive series, as well as their daughter. In addition, as it is for environmental samples, the activity of 40K should be an important source of radioactivity in all analyzed samples. Gamma rays were measured using a standard spectroscopy system, with a high-resolution HPGe detector. Measured activities in feed samples ranged from 0.23 to 1.51 Bq.kg-1, 0.67 to 4.21 Bq.kg-1, 0.29 to 1.63 Bq.kg-1 and 236 to 402 Bq.kg-1 for 226Ra, 228 Ra, 228 Th and 40K, respectively. Measured activities in DCP samples were 46.6 Bq.kg-1, 83 Bq.kg-1, 4.20 Bq.kg-1 and 16.61 Bq.kg-1 for 40 K, 226 Ra, 228 Ra and 228 Th, respectively. Although 40K activity has reached hundreds of Becquerel in feed samples, it should not represent a risk to human health, not even to animals, since potassium is an essential mineral to living organisms. <![CDATA[Contribution of the IAM Pathway to IAA Pool in Developing Rice Grains]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100507&lng=pt&nrm=iso&tlng=pt A possible role for the indole-3-acetamide (IAM) pathway in the indole-3-acetic acid (IAA) production was investigated in developing rice grains. IAM-hydrolase proposed to convert IAM to IAA primarily through the identification of IAM and IAM-hydrolase activity in some plant species. Expression profiles of the two putative rice IAM-hydrolase genes, OsAMI1&amp;2, were compared to the previously quantified IAA level. The abrupt increase in IAA level between 4 and 7 days after anthesis (DAF) was not found to correlate with changes in the expression of OsAMI1 or OsAMID2 suggesting that the IAM pathway may not contribute significantly to IAA pool in rice grains. Production of a biological compound other than IAA may explain the high activity of OsAMI1&amp;2 in developing rice grains. OsAMI1 that reported to be conserved across the plant kingdom showed higher expression level in most analyzed reproductive rice tissues whereas OsAMID2 showed more fluctuation in expression comparing to OsAMI1. Role of the IAM pathway in IAA production was also discussed in other plant systems and Arabidopsis seed was recommended as an ideal tissue to identify enzyme(s) convert(s) tryptophan to IAM as well as physiological effects of IAA produced via this pathway. <![CDATA[Blackberry Vinegar Produced By Successive Acetification Cycles: Production, Characterization And Bioactivity Parameters]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100508&lng=pt&nrm=iso&tlng=pt ABSTRACT: Blackberry vinegar was produced in successive acetification cycles and content of total phenolics, anthocyanins and antioxidant activity were evaluated along the production. Firstly, blackberry wine was obtained in bench-scale bioreactor, being verified 0.39 g/g ethanol yield, 1.78 g/L.h volumetric productivity and 76% efficiency. After, three successive acetification cycles were conducted efficiently in grapia barrel with average acetic acid production of 51.6 g/L, 72.2 % acetic acid yield and 0.4 g/L.h volumetric productivity. Appreciable contents of polyphenolic compounds, anthocyanins and high antioxidant activity were observed in the raw material, wine and vinegar obtained in each cycle of acetic acid transformation. Acetic acid transformation led the small reduction of antioxidant activity compared to alcoholic fermentation, but the antioxidant potential was maintained along the cycles. The content of total phenolics and anthocyanins also suffered a reduction in step of acetification. <![CDATA[Biological Parameters of <em>Euschistus heros</em> (F.) (Heteroptera: Pentatomidae) and its Susceptibility to Entomopathogenic Fungi When Fed on Different Diets]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100509&lng=pt&nrm=iso&tlng=pt ABSTRACT: The effects of six diets on biological parameters of Euschistus heros (F.) (Heteroptera: Pentatomidae) and its susceptibility to entomopathogenic fungi Beauveria bassiana (Bb) and Metarhizium anisopliae (Ma) were studied. The diets included a standard diet (bean pods+soybeans, peanuts and sunflower) and diets in which the bean pods were combined with only one type of grain, either soybean, peanut, sunflower, pea or sesame seeds. The best performance was that of insects fed on standard diet, which had the shortest development period and 49.5% of egg-to-adult viability. The insects fed the pea diet exhibited less than 30% viability and 11% of adults had wing deformations. The finite rate of increase values for the treatment groups were: standard: 1.091; soybean: 1.077; peanut: 1.071; sunflower: 1.076; pea: 1.058; and sesame: 1.065. The susceptibility to Ma (55%) was greater than Bb (40%) only for insects fed on the sesame diet. Considering the effects of diet on susceptibility to both fungal species comparatively among the diet-treatment groups, the stink bugs fed the restrictive diets were more susceptible compared with those fed the standard diet. We concluded that diet composition affects the biological parameters of E. heros and differentially affects their susceptibility to Bb and Ma fungi. <![CDATA[Screening and Molecular Identification of New Microbial Strains for Production of Enzymes of Biotechnological Interest]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100510&lng=pt&nrm=iso&tlng=pt ABSTRACT: This research focused on isolation, identification and characterization of new strains of fungi and bacteria, which were able to produce extracellular xylanase, mannanase, pectinase and α-amylase. Fungi isolates were identified on the basis of analyses of 18S gene sequencing and internal transcribed spacer region. The closest phylogenetic neighbors according to 18S gene sequence and ITS region data for the two isolates M1 and SE were Aspergillus fumigatus and Aspergillus sydowii, respectively. I4 was identified as Bacillus mojavensis on the basis of the 16S rRNA gene sequencing and biochemical properties. The enzyme production was evaluated by cultivating the isolated microorganisms in liquid-state bioprocess using wheat bran as carbon source. Two fungi (M1, and SE) and one bacterium (I4) strains were found to be xylanase producer, and several were proven to be outstanding producers of microbial xylanase. The strains producing xylanase secreted variable amounts of starch-debranching enzymes and produced low level β-mannan-degrading enzyme systems. The bacterium strain was found to be capable of producing pectinolytic enzymes on wheat bran at high level. Some of the strains have good potential for use as sources of important industrial enzymes. <![CDATA[Effect of Phosphorylation and Copper(II) or Iron(II) Ions Enrichment on Some Physicochemical Properties of Spelt Starch]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100511&lng=pt&nrm=iso&tlng=pt ABSTRACT: This paper provides an assessment of the effect of saturation of spelt starch and monostarch phosphate with copper or iron ions on selected physicochemical properties of the resulting modified starches. Native and modified spelt starch samples were analyzed for selected mineral element content using Atomic Absorption Spectroscopy (AAS). Thermodynamic properties were measured using DSC, and pasting properties by RVA. Flow curves of 5% pastes were plotted and described using the Herschel-Bulkley model. The structure recovery ratio was measured. AAS analysis established the presence of iron(II) and copper(II) ions in the samples of modified starches and that potassium and magnesium ions had leached from them. In comparison to unfortified samples, enriching native starch with copper(II) ions decreases value of all temperatures of phase transformation about 1.3-2.7 °C, but in case of monostarch phosphates bigger changes (2.8-3.7 °C) were observed. Fortified native spelt starch with copper(II) ions caused increasing the final viscosity of paste from 362 to 429 mPa·s. However, presence iron(II) ions in samples caused reduced its final viscosity by 170 (spelt starch) and 103 mPa·s (monostarch phosphate). Furthermore, enriching monostarch phosphate contributed to reduce degree of structure recovery of pastes from 70.9% to 66.6% in case of copper(II) ions and to 59.9% in case of iron(II) ions. <![CDATA[Value addition of corn husks through enzymatic production of xylooligosaccharides]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100512&lng=pt&nrm=iso&tlng=pt ABSTRACT Corn husks are the major wastes of corn industries with meagre economic significance. The present study was planned for value addition of corn husk through extraction of xylan, followed by its enzymatic hydrolysis into xylooligosaccharides, a pentose based prebiotic. Compositional analysis of corn husks revealed neutral detergent fibre 68.87%, acid detergent fibre 31.48%, hemicelluloses 37.39%, cellulose 29.07% and crude protein 2.68%. Irrespective of the extraction conditions, sodium hydroxide was found to be more effective in maximizing the yield of xylan from corn husks than potassium hydroxide (84% vs. 66%). Application of xylanase over the xylan of corn husks resulted into production of xylooligosaccharides with different degree of polymerization namely, xylobiose and xylotriose in addition to xylose monomer. On the basis of response surface model analysis, the maximum yield of xylobiose (1.9 mg/ml) was achieved with the enzymatic hydrolysis conditions of pH 5.8, temperature 44°C, enzyme dose 5.7U/ml and hydrolysis time of 17.5h. Therefore, the corn husks could be used as raw material for xylan extraction vis a vis its translation into prebiotic xylooligosaccharides. <![CDATA[Chemical quality of bottled mineral waters from markets of Curitiba-PR-Brazil]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100513&lng=pt&nrm=iso&tlng=pt ABSTRACT Twenty-seven bottled mineral waters from local markets in Curitiba, State of Paraná, Brazil were analyzed for a number of constituents, including: pH, conductivity, total dissolved solids, hardness, HCO3 - and CO 3 2-, as well as Ca2+, Mg2+, Na+, K+, NH4 + , F-, Cl-, NO3 -, NO2 - and SO4 2- by ion chromatography (IC); Al, Ba, Cd, Cr, Cu, Fe, Li, Mn, Ni, Sr, V and Zn by inductively coupled plasma optical emission spectrometry (ICP OES); Hg by cold vapor generation-ICP OES and As, Pb, Sb and Se by ICP-mass spectrometry (ICP-MS). The obtained limits of quantitation (LOQ) were adequate for the determination of all analytes using the various analytical techniques. Results for the analysis of AccuStandard certified reference materials QCS-01-5, QCS-02-R1-5 and TCLP-02-1 as well as spike recoveries to samples show acceptable values, within 90-111 % of expected concentrations with relative standard deviations below 10 %, demonstrating the accuracy of the determinations. Both NO2 - and NO 3 - were above the maximum limits set by Brazilian legislation in two samples, indicating a microbiological contamination. One imported sample presented As (6.1 ± 0.2 µg L-1) near the maximum limit (10 µg L-1) while all other elements of interest were below the values specified by Brazilian legislation. Principal component analysis revealed that four imported samples and one from the Minas Gerais State have the highest mineral concentrations. <![CDATA[Physicochemical and Biological Investigation of Different Structures of Carbon Coatings Deposited onto Polyurethane]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100600&lng=pt&nrm=iso&tlng=pt The aim of this study was to examine the thrombogenic properties of polyurethane that was surface modified with carbon coatings. Physicochemical properties of manufactured coatings were investigated using transmission electron microscopy (TEM), atomic force microscopy (AFM), X-ray Photoelectron Spectroscopy (XPS), Raman spectroscopy and contact angle measurement methods. Samples were examined by the Impact-R method evaluating the level of platelets activation and adhesion of particular blood cell elements. The analysis of antimicrobial resistance against E. coli colonization and viability of endothelial cells showed that polyurethane modified with use of carbon layers constituted an interesting solution for biomedical application. <![CDATA[LED Lights Enhance Metabolites and Antioxidants in Chinese Cabbage and Kale]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100601&lng=pt&nrm=iso&tlng=pt Light emitting diode (LED) lights play an important role in the plant physiology and alter the metabolites in a significant manner. Glucosinolates (GSLs), polyphenols, flavonoids and antioxidant properties of Chinese cabbage and kale cultivated in varying LED lights were investigated. Analysis revealed 7 aliphatic, 3 indolyl and 1 aromatic GSLs in Chinese cabbage and kale. The total GSL content ranged from 1.5-19.08 and 1.85-24.87 µmol/g DW, and glucobrassicanapin was the predominant GSL (3) in Chinese cabbage, whereas; sinigrin (3.49 µmol/g DW) was in kale. Blue and red LED lights produced significantly higher amount of GSLs in Chinese cabbage and kale respectively. Results revealed higher amount of total polyphenol (3.845 µg/mL) and total flavanoids (3.939 μg/mL) in Chinese cabbage. Chinese cabbage and kale showed significant antioxidant activities when compare with positive control, and the antioxidant assays were slightly correlated with total GSLs, polyphenols and flavanoids contents. The influence of LED lights on glucobrassicin in Chinese cabbage and kale should be studied extensively, because GSL is the precursor of indole-3-carbinol, a potent anticancer isothiocyanate. <![CDATA[Nanofibrous Scaffolds for Tissue Engineering Applications]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100602&lng=pt&nrm=iso&tlng=pt The discipline of tissue engineering opens up the ways for repair and regenerate damaged organs and tissues. In the current work biomimetic nanofibrous scaffolds were fabricated by electrospinning. Poly-L-lactic acid (PLLA) was blended with collagen and gelatin to fabricate PLLLA-collagen and PLLA-gelatin fibrous scaffolds respectively. Pure PLLA and gelatin scaffolds served as controls. All the scaffolds displayed randomly oriented smooth fibers studied by SEM. Surface topography and roughness were studied by AFM and surface contact angle was also measured for all the fabricated scaffolds. Surface roughness was found to be higher in collagen and gelatin blended scaffolds in comparison to PLLA scaffold. Blending of collagen and gelatin reduced the surface hydrophobicity of the scaffolds. Human osteosarcoma cell lines MG-63 were cultured on all scaffolds up to 7 days and cell adhesion was studied through SEM and confocal microscopy. SEM and confocal results showed that gelatin blended PLLA scaffold showed better cell attachment and cell spreading. <![CDATA[Investigation on chemical composition and optimization of essential oil obtainment from waste <em>Pinus taeda</em> L. using hydrodistillation]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100603&lng=pt&nrm=iso&tlng=pt ABSTRACT: The extraction of essential oils obtained by the hydrodistillation of needles/twigs waste of Pinus taeda L. was optimized by applying response surface methodology (RSM), with 24 full factorial design, in order to improve oil essential production, and aggregate value to the production chain of pine wood. Through the model it was possible to ascertain the influence of the variables in the average amount of essential oil (0.1032 mL), being the variables analyzed: biomass - Bm (x1), extraction time - ET (x2), Bm:ET (x1x2) and sample size - SS: drying times - DT (x3x4). Only linear terms (biomass and extraction time) and your interaction demonstrated significant positive values (0.0344, 0.0206 and 0.0131). The major components of the essential oil identified by GC-MS were: β-phellandrene: (30.39 and 22.44%), tricyclene (26.14 and 20.46%), β-myrcene (14.32 and 11.50%), β-pinene (22.49 and 1.43%) and α-pinene (0.25 and 11.26%) in the years 2011 and 2012, respectively. Our results show that the essential oil obtained from P. taeda represents a way of using some of the waste generated by the timber industry. The process of obtaining doesn't require treatments such as controlled drying or size reduction of the sample, indicating that it can be used in an industrial scale. <![CDATA[Late Holocene Vegetation History and Early Evidence of Araucaria angustifolia in Caçapava do Sul in the Lowland Region of Rio Grande do Sul State, Southern Brazil]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100700&lng=pt&nrm=iso&tlng=pt Little is known about the southernmost occurrence of small areas with Araucaria angustifolia populations in Caçapava do Sul in low elevated areas of Rio Grande do Sul State, about 130 km to the south of to the highlands of southern Brazil where the main distribution of Araucaria is found. This occurrence is about 130 km further south to the main area of Araucaria angustifolia which is on the highlands in southern Brazil. The question is whether this occurrence is natural, due to indigenous peoples, or due to plantation by post-Columbian settlers. To trace the origin of this little known southernmost existence of Araucaria angustifolia trees is of particular interest for conservation issues. To address this question we did a vegetation survey and studied a 150 cm-long radiocarbon dated sediment core from the Fazenda da Mônica by pollen analysis. The vegetation survey of the study area indicates that also other typical taxa of the Araucaria forest as well as the Atlantic lowland rainforest are found in the present-day semi-deciduous forest, such as Podocarpus, Ilex, Myrsine and Prunus for the former, and Alchornea, Moraceae, Arecaceae, and Myrtaceae for the later. The pollen record, due to bad pollen preservation, starts only after 44 cm core depth, which is about 515 cal yr BP old (AD 1490), indicating that Araucaria angustifolia as well as other Araucaria forest and Atlantic rainforest taxa occurred in this area since the beginning of the pollen record. The occurrence of these taxa can be seen as natural and not introduced during the post-Columbian colonisation. First settlers at the beginning of the 19th century reduced existing population of Araucaria markedly and in particular since about AD 1950. The population of Araucaria angustilfolia before the post-Columbian settlement was much larger than today. <![CDATA[Sustainable Management of Keratin Waste Biomass: Applications and Future Perspectives]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100701&lng=pt&nrm=iso&tlng=pt Keratin is a durable, fibrous protein which is mainly present in higher vertebrates (mammals, birds and reptiles) and humans epithelial cells. Food industry especially the meat market, slaughter house and wool industry produces million of tons of keratin containing biomass. These industries are constantly growing and the major producers include USA, Brazil and China, account for more than 40 million tons per year. These proteins constitute keratin by-products have from 15 to 18% nitrogen, 2-5% sulphur, 3.20% mineral elements and 1.27% fat and 90% of proteins. The organic waste rich in keratin can be utilized as a natural source using chemical and mechanical methods. The natural keratin obtained by biomass does not contain any harmful chemical and can be used directly to produce variety of cosmetics, creams, shampoos, hair conditioners and biomedical products. The natural protein is more compatible to use or apply on human skin and hairs. The monomeric units of natural keratin can penetrate in the skin and hair cuticle and able to nourish the skin without any side effects. In the present review various strategies for the purification and separation of keratin from the organic waste have been described and use of natural keratin in cosmetics and pharmaceutical industry has also been explored. <![CDATA[Heavy Metals in <strong><em>Garra gotyla</em></strong> , <strong><em>Cyprinus carpio</em></strong> and <strong><em>Cyprinion watsoni</em></strong> from the River Panjkora, District, Lower Dir, Khyber Pakhtunkhwa, Pakistan]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100702&lng=pt&nrm=iso&tlng=pt ABSTRACT This study was conducted in the River Panjkora, Pakistan, in order to assess the level of heavy metals accumulated in the liver, kidneys, gills and muscle tissues of three highly consumed fish species by the local community, Garra gotyla, Cyprinus carpio and Cyprinion watsoni. The heavy metals including manganese (Mn), iron (Fe), chromium (Cr), zinc (Zn), lead (Pb), cadmium (Cd) and nickel (Ni) were determined in the collected fish specimens through an atomic absorption spectrophotometer. Level of heavy metals significantly varied depending upon tissues type and fish species. Cyprinus carpio accumulated highest concentration of heavy metals, followed by Garra gotyla while Cyprinion watsoni accumulated least concentration of heavy metals. All the three species displayed significant differences in level of accumulated heavy metals. The order of accumulation in tissues was gills&lt;muscles&lt;kidney&lt;liver. Cd and Ni were not detected in all collected specimens while Mn was not detected in the tissues of Cyprinion watsoni collected from the reference site. <![CDATA[Land Use and Environmental Zoning of Mixed Ombrophilous Forests for Sustainable Use (Irati National Forest, Brazil Southern Region)]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100703&lng=pt&nrm=iso&tlng=pt ABSTRACT An environmental zoning proposal due to the demand to develop strategies to ensure biodiversity scenarios for the remaining mixed ombrophilous forests was carried out for the Irati National Forest, a legally protected area for sustainable use located in southern Brazil. To identify land use and cover, Ikonos PSM images dated from December 2004, based on the interpretation of visual character and vectorization of orbital data were used. Qualitative and quantitative classifications identified six types of land and cover use: native forest, forestry, capoeira grass, wetland, araucaria plantation, and anthropic. The native forest relative to the mixed ombrophilous forest was the prevailing type of land cover. The environmental zoning proposal defined seven zones of use, some that involved several potentially harmful activities to the biodiversity of the Irati National Forest. The environmental zone proposal, based on ecosystem conservation, outlined specific factors that enhanced ecosystem and community viability, supporting the design and evaluation of the management plan of the Irati National Forest. <![CDATA[Erratum: Effect of LPS on the Viability and Proliferation of Human Oral and Esophageal Cancer Cell Lines]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100901&lng=pt&nrm=iso&tlng=pt ABSTRACT An environmental zoning proposal due to the demand to develop strategies to ensure biodiversity scenarios for the remaining mixed ombrophilous forests was carried out for the Irati National Forest, a legally protected area for sustainable use located in southern Brazil. To identify land use and cover, Ikonos PSM images dated from December 2004, based on the interpretation of visual character and vectorization of orbital data were used. Qualitative and quantitative classifications identified six types of land and cover use: native forest, forestry, capoeira grass, wetland, araucaria plantation, and anthropic. The native forest relative to the mixed ombrophilous forest was the prevailing type of land cover. The environmental zoning proposal defined seven zones of use, some that involved several potentially harmful activities to the biodiversity of the Irati National Forest. The environmental zone proposal, based on ecosystem conservation, outlined specific factors that enhanced ecosystem and community viability, supporting the design and evaluation of the management plan of the Irati National Forest. <![CDATA[Erratum: Regulating Effect Of Carnosine And /Or L- Arginine On The Expression Of Inflammatory Molecules Induced Nephropathy In The Hypoxic Rat Model]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100902&lng=pt&nrm=iso&tlng=pt ABSTRACT An environmental zoning proposal due to the demand to develop strategies to ensure biodiversity scenarios for the remaining mixed ombrophilous forests was carried out for the Irati National Forest, a legally protected area for sustainable use located in southern Brazil. To identify land use and cover, Ikonos PSM images dated from December 2004, based on the interpretation of visual character and vectorization of orbital data were used. Qualitative and quantitative classifications identified six types of land and cover use: native forest, forestry, capoeira grass, wetland, araucaria plantation, and anthropic. The native forest relative to the mixed ombrophilous forest was the prevailing type of land cover. The environmental zoning proposal defined seven zones of use, some that involved several potentially harmful activities to the biodiversity of the Irati National Forest. The environmental zone proposal, based on ecosystem conservation, outlined specific factors that enhanced ecosystem and community viability, supporting the design and evaluation of the management plan of the Irati National Forest. <![CDATA[Erratum: Recurrent Somatic Embryogenesis and Plantlet Regeneration in <em>Psidium</em> guajava L.]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100903&lng=pt&nrm=iso&tlng=pt ABSTRACT An environmental zoning proposal due to the demand to develop strategies to ensure biodiversity scenarios for the remaining mixed ombrophilous forests was carried out for the Irati National Forest, a legally protected area for sustainable use located in southern Brazil. To identify land use and cover, Ikonos PSM images dated from December 2004, based on the interpretation of visual character and vectorization of orbital data were used. Qualitative and quantitative classifications identified six types of land and cover use: native forest, forestry, capoeira grass, wetland, araucaria plantation, and anthropic. The native forest relative to the mixed ombrophilous forest was the prevailing type of land cover. The environmental zoning proposal defined seven zones of use, some that involved several potentially harmful activities to the biodiversity of the Irati National Forest. The environmental zone proposal, based on ecosystem conservation, outlined specific factors that enhanced ecosystem and community viability, supporting the design and evaluation of the management plan of the Irati National Forest. <![CDATA[Erratum: Effects of Sodium Nitrate and Mixotrophic Culture on Biomass and Lipid Production in Hypersaline Microalgae <em>Dunaliella Viridis</em> Teod]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100904&lng=pt&nrm=iso&tlng=pt ABSTRACT An environmental zoning proposal due to the demand to develop strategies to ensure biodiversity scenarios for the remaining mixed ombrophilous forests was carried out for the Irati National Forest, a legally protected area for sustainable use located in southern Brazil. To identify land use and cover, Ikonos PSM images dated from December 2004, based on the interpretation of visual character and vectorization of orbital data were used. Qualitative and quantitative classifications identified six types of land and cover use: native forest, forestry, capoeira grass, wetland, araucaria plantation, and anthropic. The native forest relative to the mixed ombrophilous forest was the prevailing type of land cover. The environmental zoning proposal defined seven zones of use, some that involved several potentially harmful activities to the biodiversity of the Irati National Forest. The environmental zone proposal, based on ecosystem conservation, outlined specific factors that enhanced ecosystem and community viability, supporting the design and evaluation of the management plan of the Irati National Forest. <![CDATA[Erratum:Development of an L-Lysine Enriched Bran for Animal Nutrition via Submerged Fermentation by Corynebacterium glutamicum using Agroindustrial Substrates]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100905&lng=pt&nrm=iso&tlng=pt ABSTRACT An environmental zoning proposal due to the demand to develop strategies to ensure biodiversity scenarios for the remaining mixed ombrophilous forests was carried out for the Irati National Forest, a legally protected area for sustainable use located in southern Brazil. To identify land use and cover, Ikonos PSM images dated from December 2004, based on the interpretation of visual character and vectorization of orbital data were used. Qualitative and quantitative classifications identified six types of land and cover use: native forest, forestry, capoeira grass, wetland, araucaria plantation, and anthropic. The native forest relative to the mixed ombrophilous forest was the prevailing type of land cover. The environmental zoning proposal defined seven zones of use, some that involved several potentially harmful activities to the biodiversity of the Irati National Forest. The environmental zone proposal, based on ecosystem conservation, outlined specific factors that enhanced ecosystem and community viability, supporting the design and evaluation of the management plan of the Irati National Forest. <![CDATA[Erratum:Contribution of the IAM Pathway to IAA Pool in Developing Rice Grains]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100906&lng=pt&nrm=iso&tlng=pt ABSTRACT An environmental zoning proposal due to the demand to develop strategies to ensure biodiversity scenarios for the remaining mixed ombrophilous forests was carried out for the Irati National Forest, a legally protected area for sustainable use located in southern Brazil. To identify land use and cover, Ikonos PSM images dated from December 2004, based on the interpretation of visual character and vectorization of orbital data were used. Qualitative and quantitative classifications identified six types of land and cover use: native forest, forestry, capoeira grass, wetland, araucaria plantation, and anthropic. The native forest relative to the mixed ombrophilous forest was the prevailing type of land cover. The environmental zoning proposal defined seven zones of use, some that involved several potentially harmful activities to the biodiversity of the Irati National Forest. The environmental zone proposal, based on ecosystem conservation, outlined specific factors that enhanced ecosystem and community viability, supporting the design and evaluation of the management plan of the Irati National Forest. <![CDATA[Erratum: Investigation of the <em>GJB6</em> Deletion Mutations Del (GJB6-D13s1830) and Del (GJB6-D13s1854) in Iranian Patients with Autosomal-Recessive Non-Syndromic Hearing Loss (ARNSHL)]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100907&lng=pt&nrm=iso&tlng=pt ABSTRACT An environmental zoning proposal due to the demand to develop strategies to ensure biodiversity scenarios for the remaining mixed ombrophilous forests was carried out for the Irati National Forest, a legally protected area for sustainable use located in southern Brazil. To identify land use and cover, Ikonos PSM images dated from December 2004, based on the interpretation of visual character and vectorization of orbital data were used. Qualitative and quantitative classifications identified six types of land and cover use: native forest, forestry, capoeira grass, wetland, araucaria plantation, and anthropic. The native forest relative to the mixed ombrophilous forest was the prevailing type of land cover. The environmental zoning proposal defined seven zones of use, some that involved several potentially harmful activities to the biodiversity of the Irati National Forest. The environmental zone proposal, based on ecosystem conservation, outlined specific factors that enhanced ecosystem and community viability, supporting the design and evaluation of the management plan of the Irati National Forest. <![CDATA[Erratum: Effect of Gamma Radiation on Growth and Metabolic Activities of <em>Arthrospira platensis</em>]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100908&lng=pt&nrm=iso&tlng=pt ABSTRACT An environmental zoning proposal due to the demand to develop strategies to ensure biodiversity scenarios for the remaining mixed ombrophilous forests was carried out for the Irati National Forest, a legally protected area for sustainable use located in southern Brazil. To identify land use and cover, Ikonos PSM images dated from December 2004, based on the interpretation of visual character and vectorization of orbital data were used. Qualitative and quantitative classifications identified six types of land and cover use: native forest, forestry, capoeira grass, wetland, araucaria plantation, and anthropic. The native forest relative to the mixed ombrophilous forest was the prevailing type of land cover. The environmental zoning proposal defined seven zones of use, some that involved several potentially harmful activities to the biodiversity of the Irati National Forest. The environmental zone proposal, based on ecosystem conservation, outlined specific factors that enhanced ecosystem and community viability, supporting the design and evaluation of the management plan of the Irati National Forest. <![CDATA[Erratum: Nanofibrous Scaffolds for Tissue Engineering Applications]]> http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100909&lng=pt&nrm=iso&tlng=pt ABSTRACT An environmental zoning proposal due to the demand to develop strategies to ensure biodiversity scenarios for the remaining mixed ombrophilous forests was carried out for the Irati National Forest, a legally protected area for sustainable use located in southern Brazil. To identify land use and cover, Ikonos PSM images dated from December 2004, based on the interpretation of visual character and vectorization of orbital data were used. Qualitative and quantitative classifications identified six types of land and cover use: native forest, forestry, capoeira grass, wetland, araucaria plantation, and anthropic. The native forest relative to the mixed ombrophilous forest was the prevailing type of land cover. The environmental zoning proposal defined seven zones of use, some that involved several potentially harmful activities to the biodiversity of the Irati National Forest. The environmental zone proposal, based on ecosystem conservation, outlined specific factors that enhanced ecosystem and community viability, supporting the design and evaluation of the management plan of the Irati National Forest.