Primjena metode odzivnih površina radi optimiranja proizvodnje mliječne kiseline: obogaćivanje podloge, kontrola temperature i pH-vrijednosti

Two response surface methodologies involving central composite designs have been successfully applied to evaluate the effect of cheese whey, corn steep liquor, ammonium sulphate, temperature and pH control on lactic acid fermentation by Lactobacillus sp. LMI8 isolated from cassava flour wastewater. In the first central composite design, corn steep liquor and ammonium sulphate were investigated as low-cost nitrogen sources in combination with other components to substitute yeast extract for economical production. The best results were obtained with 55 g/L of lactose, 15 g/L of corn steep liquor and 5.625 g/L of ammonium sulphate. At the maximum point, the lactic acid concentration reached 18.68 g/L. After defining the optimal nutritional conditions for lactic acid production, a second central composite design was performed to determine the extent to which temperature and pH influence the lactic acid production with the aim of improving the fermentation process. The second-order polynomial regression model determined that the maximum lactic acid production of 52.37 g/L would be obtained when the optimum temperature and pH were 39.6 °C and 5.9, respectively. Comparing the lactic acid production in shake flask fermentation, there was an increase of 180 % after 30 h of processing, with a conversion efficiency of about 86.12 % of the initial lactose. In addition, lactic acid produced from whey lactose by Lactobacillus sp. LMI8 was optically almost pure D-lactic acid (over 98 % of total lactic acid produced).U radu su uspješno primijenjene dvije metode odzivnih površina, uključujući centralno složeni dizajn, za procjenu učinka koncentracije sirutke, ekstrakta kukuruza, amonijeva sulfata, temperature i pH-vrijednosti na mliječno-kiselu fermentaciju uz pomoć bakterije Lactobacillus sp. LMI8, izolirane iz otpadnih voda nakon proizvodnje brašna kasave. U prvom centralno složenom dizajnu ispitani su ekstrakt kukuruza i amonijev sulfat kao jeftini izvori dušika. Utvrđeno je da se u kombinaciji s ostalim sastojcima mogu upotrijebiti kao ekonomični nadomjestak za kvaščev ekstrakt. Najbolji su rezultati postignuti uporabom 55 g/L laktoze, 15 g/L ekstrakta kukuruza i 5,625 g/L amonijeva sulfata, pri čemu je proizvedeno 18,68 g/L mliječne kiseline. Nakon što je određen optimalni sastav hranjiva, proveden je drugi složeno centralno složeni dizajn kako bi se ispitao utjecaj temperature i pH-vrijednosti na proizvodnju mliječne kiseline i poboljšao proces fermentacije. Polinomskim regresijskim modelom drugoga reda određeno je da se maksimalna proizvodnja mliječne kiseline od 52,37 g/L može postići pri temperaturi od 39,6 °C i pH-vrijednosti od 5,9. U usporedbi s fermentacijom na tresilici, koncentracija mliječne kiseline nakon 30 h bila je 180 % veća, a uspješnost konverzije laktoze iznosila 86,12 %. Također je više od 98 % mliječne kiseline proizvedene iz laktoze sirutke s pomoću Lactobacillus sp. LMI8 bila skoro optički čista D-mliječna kiselina

Moringa oleifera: Resource management and multiuse life tree

Moringa oleifera Lamarck (Moringaceae family) is a plant native from the Western and sub-Himalayan parts of Northwest India, Pakistan and Afghanistan. This species is widely cultivated across Africa, South-East Asia, Arabia, South America and Caribbean Islands. M. oleifera culture is also being distributed in the Semi-Arid Northeast of Brazil. It is a multiuse life tree with great environmental economic importance in industrial and medical areas. This review reports different purposes of M. oleifera including sustaining environmental resources, soil protection and shelter for animals. This plant requires not much care and distinct parts have bioactive compounds. Moringa tissues used in human and animal diets, also withdraw pollutants from water. The seeds with coagulant properties used in water treatment for human consumption, remove waste products like surfactants, heavy metals and pesticides. The oil extracted from seeds is used in cosmetic production and as biodiesel. M. oleifera tissues also contain proteins with different biological activities, including lectins, chitin-binding proteins, trypsin inhibitors, and proteases. The lectins are reported to act as insecticidal agents against Aedes aegypti (vector of dengue, chikungunya and yellow fevers) and Anagasta kuehniella (pest of stored products) and also showed water coagulant, antibacterial and blood anticoagulant activities. The presence of trypsin inhibitors has been reported in M. oleifera leaves and flowers. The inhibitor from flowers is toxic to larvae of A. aegypti. The flowers also contain caseinolytic proteases that are able to promote clotting of milk. In this sense, M. oleifera is a promising tree from a biotechnological point of view, since it has shown a great variety of uses and it is a source of several compounds with a broad range of biological activities.Conselho Nacional de Desenvolvimento Científico e Tecnológico for fellowship (LCBBC) and to the Foundation for Science and Technology, POPH/FSE (AFSS

Isolation of a seed coagulant Moringa oleifera lectin

In this work hemagglutinating activity (HA) was investigated in distinct Moringa oleifera tissue extracts. A new lectin from seeds (cMoL) was purified and characterized; hemagglutinating and coagulating activities were evaluated. HA was detected in 0.15 M NaCl extracts from flowers and rachis inflorescence (5%, w/v), seeds, leaves, fundamental tissue of stem and steam bark (10%, w/v). cMoL isolated after saline extraction and guar gel column chromatography was active at pH range 4.0–9.0 agglutinating erythrocytes from rabbit and human blood types. Extracts of tissues and cMoL activities were carbohydrate inhibited; azocasein and asialofetuin abolished cMoL HA. The lectin was thermostable at 100 °C during 7 h. Polyacrylamide gel electrophoresis under reduced conditions revealed a main polypeptide band of 26.5 kDa; native basic cMoL was detected as a unique band. Seed lectin preparations and cMoL showed coagulant activity, similar to aluminium sulphate, the coagulant most widely used in water treatment.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco (FACEPE)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Científico e Tecnológico (CNPq

Tissue distribution of DNA-Hsp65/TDM-loaded PLGA microspheres and uptake by phagocytic cells

This study aimed to demonstrate that microspheres, used as delivery vehicle of DNA-Hsp65/TDM [plasmid DNA encoding heat shock protein 65 (Hsp65) coencapsulated with trehalose dimycolate (TDM) into PLGA microspheres], are widely spread among several organs after intramuscular administration in BALB/c mice. In general, we showed that these particles were phagocytosed by antigen presenting cells, such as macrophages and dendritic cells. Besides, it was demonstrated herein that draining lymph node cells presented a significant increase in the number of cells expressing costimulatory molecules (CD80 and CD86) and MHC class II, and also that the administration of the DNA-Hsp65/TDM and vector/TDM formulations resulted in the up-regulation of CD80, CD86 and MHC class II expression when compared to control formulations (vector/TDM and empty). Regarding the intracellular trafficking we observed that following phagocytosis, the microspheres were not found in the late endosomes and/or lysosomes, until 15 days after internalization, and we suggest that these constructions were hydrolysed in early compartments. Overall, these data expand our knowledge on PLGA [poly (lactic-co- glycolic acid)] microspheres as gene carriers in vaccination strategies, as well as open perspectives for their potential use in clinical practice

Moringa oleifera lectin and binding affinity to humic acids : application to water treatment

Humic acids, natural organic matter, constitute a problem in water treatment; they react with chlorine forming undesirable disinfection byproducts. The aim of this work was the purification of a saline soluble Moringa oleifera lectin (SSMoL) and characterization of its affinity to humic acids. SSMoL isolation included extraction of seed flour, saline fractionation and guar gel chromatography. Hemagglutinating activity (HA) of extract (E), fraction (0-60F) and SSMoL was assessed with rabbit erythrocytes. HA inhibitions were done with humic acid, carbohydrates, glycoproteins and halogenated organic compounds. Diffusion experiments with humic acid and lectin preparations were performed in agarose gel. Basic SSMoL showed two polypeptide bands with ß-mercapthoethanol treatment. SSMoL HA with humic acid decreased from 512-1 to 32-1. Similar results were obtained with E and 0-60F. SSMoL was partially inhibited by carbohydrates and glycoproteins; azocasein abolished SSMoL activity. SSMoL HA was not altered with trichloroacetic and dicholoroacetic acids, or chloroform. Precipitation bands were observed in diffusion gel. The present study foresees a future application of SSMoL in water treatment to remove humic acids

B cells Can Modulate the CD8 Memory T Cell after DNA Vaccination Against Experimental Tuberculosis

Abstract\ud \ud \ud \ud Background\ud \ud Although B cells are important as antigen presenting cells (APC) during the immune response, their role in DNA vaccination models is unknown.\ud \ud \ud \ud Methods\ud \ud In this study in vitro and in vivo experiments were performed to evaluate the ability of B cells to protect mice against Mycobacterium tuberculosis challenge.\ud \ud \ud \ud Results\ud \ud \ud In vitro and in vivo studies showed that B cells efficiently present antigens after naked plasmid pcDNA3 encoding M. leprae 65-kDa heat shock protein (pcDNA3-Hsp65) internalization and protect B knock-out (BKO) mice against Mycobacterium tuberculosis infection. pcDNA3-Hsp65-transfected B cells adoptively transferred into BKO mice rescued the memory phenotypes and reduced the number of CFU compared to wild-type mice.\ud \ud \ud \ud Conclusions\ud \ud These data not only suggest that B cells play an important role in the induction of CD8 T cells but also that they improve bacterial clearance in DNA vaccine model.We are thankful to Ana Paula Masson and Izaíra T Brandão for providing the DNA vaccine and recombinant protein. This study was supported by a FAPESP fellowship (05/030873) to LPA.We are thankful to Ana Paula Masson and Izaíra T Brandão for providing the DNA vaccine and recombinant protein. This study was supported by a FAPESP fellowship (05/03087-3) to LPA

MLL5 improves ATRA driven differentiation and promotes xenotransplant engraftment in acute promyelocytic leukemia model

Although the mixed lineage leukemia 5 (MLL5) gene has prognostic implications in acute promyelocyte leukemia (APL), the underlying mechanism remains to be elucidated. Here, we demonstrate the critical role exerted by MLL5 in APL regarding cell proliferation and resistance to drug-induced apoptosis, through mtROS regulation. Additionally, MLL5 overexpression increased the responsiveness of APL leukemic cells to all-trans retinoic acid (ATRA)-induced differentiation, via regulation of the epigenetic modifiers SETD7 and LSD1. In silico analysis indicated that APL blasts with MLL5(high) transcript levels were associated with retinoic acid binding and downstream signaling, while MLL5(low) blasts displayed decreased expression of epigenetic modifiers (such as KMT2C, PHF8 and ARID4A). Finally, APL xenograft transplants demonstrated improved engraftment of MLL5-expressing cells and increased myeloid differentiation over time. Concordantly, evaluation of engrafted blasts revealed increased responsiveness of MLL5-expressing cells to ATRA-induced granulocytic differentiation. Together, we describe the epigenetic changes triggered by the interaction of MLL5 and ATRA resulting in enhanced granulocytic differentiation

Preparação de lectinas de Moringa oleifera com atividade coagulante

info:eu-repo/semantics/publishedVersio