Scielo RSS <![CDATA[Brazilian Archives of Biology and Technology]]> vol. 59 num. lang. pt <![CDATA[SciELO Logo]]> <![CDATA[Use of Solid Waste from Thermoelectric Plants for the Cultivation of Microalgae]]> 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]]> 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[Expression of Ghrelin and GHSR-1a in Long Term Diabetic Rat's Kidney]]> 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)]]> 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]]> 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]]> 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]]> 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]]> 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]]> 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]]> 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]]> 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]]> 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]]> 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 GJB6 Deletion Mutations Del (GJB6-D13s1830) and Del (GJB6-D13s1854) in Iranian Patients with Autosomal-Recessive Non-Syndromic Hearing Loss (ARNSHL)]]> 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]]> 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)]]> 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]]> 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[Optimization of Lipase-Catalyzed Transesterification of Cotton Seed Oil for Biodiesel Production Using Response Surface Methodology]]> 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]]> 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]]> 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.]]> 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 Jatropha curcas L During Development and Under Stress Conditions Using Real Time Quantitative PCR]]> 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]]> 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]]> 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)]]> 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]]> 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 Arthrospira Platensis]]> 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]]> 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]]> 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[Spray Drying of Pequi Pulp: Process Performance and Physicochemical and Nutritional Properties of the Powdered Pulp]]> 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>]]> 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]]> 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]]> 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[Physicochemical and Biological Investigation of Different Structures of Carbon Coatings Deposited onto Polyurethane]]> 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]]> 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[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]]> 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]]> 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.