Brewery effluents supplemented with sugarcane molasses and urea were used as growth medium to develop the yeast Rhodosporidium toruloires NCYC 921, to produce triacylglicerols (TAGs) and carotenoids. ...The yeast cultivation was monitored by flow cytometry, in order to understand the impact of the effluent on the yeast cell membrane. Ideally the treated effluent may be discharged in the water bodies.
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•Brewery effluent was used to grow R. toruloides for lipids and carotenoids production.•Sugarcane molasses and urea improved the yeast lipids and carotenoids production.•Flow cytometry was successfully used to monitor the yeast cell membrane integrity.
In this study, secondary brewery wastewater (SBWW) supplemented with sugarcane molasses (SCM) was used for SBWW treatment with concomitant lipid and carotenoid production by the yeast Rhodosporidium toruloides NCYC 921. In order to improve the biomass production, ammonium sulfate, yeast extract and urea were tested as nitrogen sources. Urea was chosen as the best low-cost nitrogen source. A fed-batch cultivation was carried out with SBWW supplemented with 10 g L−1 of sugarcane molasses as carbon source, and 2 g L−1 of urea as nitrogen source. A maximum biomass concentration of 42.5 g L−1 was obtained at t = 126.5 h and the maximum biomass productivity was 0.55 g L−1 h−1 at t = 48.25 h. The maximum lipid content was 29.9 % w/w (DCW) at t = 94 h of cultivation and the maximum carotenoid content was 0.23 mg g−1 at 120 h of cultivation. Relatively to the SBWW treatment, after the batch phase, 45.8 % of total Kjeldahl nitrogen removal, 81.7 % of COD removal and 100 % of sugar consumption were observed. Flow cytometry analysis revealed that 27.27 % of the cells had injured membrane after the inoculation. This proportion was reduced to 10.37 % at the end of the cultivation, indicating that cells adapted to the growth conditions.
In most X-ray diffraction studies reported vanadate binds as a 5-coordinate moiety within the protein.
•Interactions of vanadium compounds with proteins are reviewed.•Structural and functional ...aspects of vanadium–protein interactions are discussed.•The interactions of vanadium ions with enzymes may imply interference at distinct levels.•Phosphate–vanadate physicochemical similarities are relevant in protein binding and inhibition.•Vanadate(IV) can also mimic phosphate.
Vanadium is an element ubiquitously present in our planet's crust and thus there are several organisms that use vanadium for activity or function of proteins. Examples are the vanadium-dependent haloperoxidases and the vanadium-containing nitrogenases. Some organisms that use vanadium have extremely efficient and selective protein-dependent systems for uptake and transport of vanadium and are able to accumulate high levels of vanadium from seawater, vanabins being a unique family of vanadium binding proteins found in ascidians involved in this process. For all of the systems a discussion regarding the role of the V-containing proteins is provided, mostly centered on structural aspects of the vanadium site and, when possible or relevant, relating this to the mechanisms operating. Phosphate is very important in biological systems and is involved in an extensive number of biological recognition and bio-catalytic systems. Vanadate(V) is able to inhibit many of the enzymes involved in these processes, such as ATPases, phosphatases, ribonucleases, phosphodiesterases, phosphoglucomutase and glucose-6-phosphatase, and it appears clear that this is closely related to the analogous physicochemical properties of vanadate and phosphate. The ability of vanadium to interfere with the metabolic processes involving Ca2+ and Mg2+, connected with its versatility to undergo changes in coordination geometry, allow V to influence the function of a large variety of phosphate-metabolizing enzymes and vanadate(V) salts and compounds have been frequently used either as inhibitors of these enzymes, or as probes to study the mechanisms of their reactions and catalytic cycle. In this review we give an overview of the many examples so far reported, also disclosing that vanadate(IV) may also have an equally efficient inhibiting effect. The prospective application of vanadium compounds as therapeutics has also been an important topic of research. How vanadium may be transported in blood and up-taken by cells are particularly relevant issues, this being mainly dependent on transferrin (and albumin) present in blood plasma. The thousands of studies reported on the effects of vanadium compounds reflect the complexity of the interactions occurring. Although it is not easy to anticipate/determine if a particular effect observed in a test tube or in vitro is also going to take place in vivo, it is clear that vanadium ions may interfere with many metabolic processes at many distinct levels. Emphasis is given on structural and functional aspects of vanadium–protein interactions relevant for vanadium binding and/or for clarification of role of the metal center in the reaction mechanisms. The additional knowledge that the presence of vanadium can change the action of a protein, other than simply inhibiting it, may also be important to understand how vanadium affects biological systems. This possibility, together with the vanadate–phosphate analogy further potentiates the belief that vanadium probably has relevant functions in living beings, which may involve interaction or incorporation of the metal ion and/or its compounds with several proteins.
Recently, yeast and microalgae mixed cultures have been widely used in biological effluent treatments and biofuel production because such cultures show many advantages over pure cultures. However, ...industrial effluents often contain toxic compounds; therefore, it is important to evaluate the cell stress response when growing in such conditions during the mixed culture development. In this work, flow cytometry (FC) was used to differentiate
Rhodosporidium toruloides
cells from
Tetradesmus obliquus
cells, based on their size, internal complexity, and chlorophyll content. FC coupled with SYTOX Green and CFDA fluorochromes was also used to characterize the cell stress response of
R. toruloides
and
T. obliquus
individual cells in a mixed culture. This work describes, for the first time, a simple and easy method to monitor individual stress response of
R. toruloides
and
T. obliquus
cells growing in mixed cultures on brewery effluents, using FC coupled with fluorescent dyes.
The induced expression of nitric oxide synthase (iNOS) controls the intracellular growth of Leishmania in infected macrophages. Histones deacetylases (HDACs) negatively regulate gene expression ...through the formation of complexes containing transcription factors such as NF‐κB p50/50. Herein, we demonstrated the occupancy of p50/p50_HDAC1 to iNOS promoter associated with reduced levels of H3K9Ac. Remarkably, we found increased levels of HDAC1 in L. amazonensis‐infected macrophages. HDAC1 upregulation was not found in L. major‐infected macrophages. The parasite intracellular load was reduced in HDAC1 knocked‐down macrophages, which presented increased nitric oxide levels. HDAC1 silencing led to the occupancy of CBP/p300 to iNOS promoter and the rise of H3K9Ac modification. Importantly, the immunostaining of skin samples from hiporeactive cutaneous leishmaniasis patients infected with L. amazonensis, revealed high levels of HDAC1. In brief, L. amazonensis induces HDAC1 in infected macrophages, which contribute to parasite survival and is associated to hiporeactive stage found in L. amazonensis infected patients.
Leishmania amazonensis infection induces NF‐κB p50/p50 and increases the nuclear levels of HDAC1 in macrophages. In the nucleus, p50/50 binds to nk promoter sequences and recruits HDAC1, forming a repression complex leading to H3K9 deacetylation and consequent iNOS promoter silencing. The absence of Nitric Oxide production favors the amastigote growth.
BackgroundFibromyalgia is a syndrome of chronic, generalized muscular pain, accompanied by sleep disturbances, fatigue and cardic autonomic dysfunction that will affect the quality of life. There is ...currently no gold standard treatment. There are limitations of studies with electroacupuncture in auricular acupuncture. ObjectivesWe evaluate the effects of systemic electroacupuncture (EA) with frequencies of 2/100 Hz associated of auricular acupuncture with a Nogier frequency (2.28, 4.56 and 9.12 Hz) for pain intensity, heart rate variability (HRV), and quality of life in fibromyalgia. MethodsRandomized clinical trial, a pilot study. Eighteen volunteers were randomized into a control group (CG, n = 9) and an experimental group (EG, n = 9). Six systemic EA sessions systemic and auricular were applied in the EG for 20 min, twice a week, for six weeks consecutive. The Numerical Pain Assessment Scale (NPRS), 2010 diagnostic criteria of the American College of Rheumatology (FDC 2010), Fibromyalgia Impact Questionnaire (FIQ) and analysis of HRV were the instruments used. The independent t-test compared to the groups was applied. ResultsThere was no statistically significant difference for the primary outcome for NPRS (p > 0.05). In the secondary outcome there was a significant difference in the total score and in some FIQ domains (p = 0.008) and some variables such as pain (p = 0.02) and anxiety (p = 0.006). There was no significant difference for the FDC 2010 and HRV variables (p > 0.05). Conclusion2/100 Hz systemic EA associated with the Nogier frequency positively influenced some quality of life variables; however, pain intensity, diagnostic criteria, and HRV variables did not change.
Brewery wastewater has been proposed as an attractive low-cost substrate for microbial lipid production for oleaginous yeast and microalga with promising results. For each liter of beer produced, ...from 3 to 10 L of wastewater are generated which can be used as culture medium for autotrophic or heterotrophic metabolism. This strategy allows reducing the culture medium cost, as well as obtaining high lipid contents and other high value compounds which can make the process profitable. Additionally, the use of industrial effluents/wastes as substrates for microbial growth can be a strategy to treat them based on the circular economy rules. This review presents the different brewery wastewater treatment strategies using oleaginous yeast and microalga pure and mixed cultures for the concomitant wastewater treatment and lipids/carotenoids production so far reported, highlighting the benefits/disadvantages of such strategies and comparing their performance in terms of wastewater treatment, lipids and carotenoids production between pure and mixed cultures performance.
Graphical Abstract
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•R. toruloides and T. obliquus pure and mixed cultures were grown on PBWW.•R. toruloides did not grow in pure cultures but grew in mixed cultures.•T. obliquus pure cultures grew on ...PBWW attaining higher DCW than in mixed cultures.•PBWW supplemented with sugarcane molasses and urea was used to grow mixed cultures.•A maximum lipid content of 26.3 % (w/w DCW) when PBWW + urea+100 g/l SCM was used.
Rhodosporidium toruloides and Tetradesmus obliquus pure and mixed cultures were grown on primary brewery wastewater (PBWW), with and without supplementation of sugarcane molasses (SCM) and urea. R. toruloides developed in pure cultures was not able to grow on PBWW, with or without supplementation, but grew in mixed cultures with the microalga T. obliquus in all media. In contrast, all T. obliquus pure cultures developed on PBWW, with and without supplementation, were able to develop. Higher biomass productivity (149.3 mg L−1 h−1) was obtained for the mixed culture using PBWW supplemented with 10 g L−1 of SCM and 2 g L−1 of urea. Therefore, a strategy to stimulate lipid production by R. toruloides and T. obliquus grown in mixed cultures with higher SCM concentrations (20, 40 and 100 g L-1 of SCM) was developed. The maximum lipid content (26.3 % (w/w DCW)) was obtained for the mixed culture developed on PBWW, supplemented with 100 g L-1 of SCM plus 2 g L-1 of urea, at t = 121 h. This work demonstrated the feasibility of using PBWW supplemented with SCM and urea as culture medium to grow R. toruloides and T. obliquus and to produce microbial lipids.
Brewery wastewater (BWW) is an appealing low-cost substrate for the production of single cell oils by oleaginous microorganisms. However, it may contain inhibitor compounds that may affect the ...microbial metabolism. This work investigated, for the first time, the presence of potential inhibitor compounds in primary brewery wastewater (PBWW) and secondary brewery wastewater (SBWW) for the pure and mixed cultivation of the yeast Rhodosporidium toruloides NCYC 921 and the microalga Tetradesmus obliquus ACOI 204/07. Three organic acids (OrgAc) were identified in the brewery effluents (acetic, propionic and butyric acids). Yeast and microalga pure and mixed cultivations were performed in PBWW and SBWW in order to understand the behaviour of the microorganisms, individually and together. Flow cytometry (FC) was used to monitor each microbial population during the mixed cultivations, and to study the yeast and microalga cell viability throughout all cultivations. The yeast cells in pure cultures grown in both effluents were severely affected by the OrgAc presence confirmed by the cell stress results obtained by FC. However, in the mixed cultures, the yeast cells were able to develop, and the levels of stress conditions were considerably lower. Only in microalga pure and mixed cultures efficient OrgAc removal was observed.
•Brewery wastewater (BWW) contains inhibitors that may affect the microbial metabolism.•It was possible to identify the presence of three organic acids (OrgAc) in BWW.•R. toruloides and T. obliquus pure and mixed cultivations were performed using BWW.•R. toruloides pure cultures were severely affected by the OrgAc present in the BWW.•T. obliquus pure and mixed cultures were efficient in removing the OrgAc in the BWW.
Biodiesel produced from oleaginous microorganisms is considered an appealing option since oleaginous yeasts and microalgae can accumulate between 20 % and 80 % lipids of their dry cell weight. ...However, it is still not economically sustainable since production costs are still high. This work developed a new mild protocol using hexane or sunflower oil as extraction solvents to simultaneously extract lipids and carotenoids from Rhodosporidium toruloides biomass directly from the broth culture without using any harvesting technique after performing high pressure homogenization to disrupt the yeast cells. The highest results were obtained for the hexane extractions attaining 55.9 % of total lipids (w/w) and 111.5 µg g−1 (eq. torulene) of carotenoids, which are higher values than the ones obtained using lipid extraction traditional methods. The process here presented is an easy, simple, inexpensive and environmentally friendly methodology which involves the simultaneous extraction of carotenoids and lipids directly from R. toruloides broth culture which has never been reported before and can greatly improve the economics of the biodiesel production process.
•A new protocol to extract lipids and carotenoids from R. toruloides was developed.•No harvesting technique was performed after high pressure homogenization.•Hexane extractions attained 55.9 % of total lipids and 111.5 µg g−1 of carotenoids.•Easy, simple and inexpensive method for simultaneously extract lipids and carotenoids.•This process can greatly improve the economics of the biodiesel production process.
Leishmania parasites are transmitted to vertebrate hosts by phlebotomine sandflies and, in humans, may cause tegumentary or visceral leishmaniasis. The role of PKR (dsRNA activated kinase) and ...Toll-like receptor 3 (TLR3) activation in the control of Leishmania infection highlights the importance of the engagement of RNA sensors, which are usually involved in the antiviral cell response, in the fate of parasitism by Leishmania. We tested the hypothesis that Phlebovirus, a subgroup of the Bunyaviridae, transmitted by sandflies, would interfere with Leishmania infection.
We tested two Phlebovirus isolates, Icoaraci and Pacui, from the rodents Nectomys sp. and Oryzomys sp., respectively, both natural sylvatic reservoir of Leishmania (Leishmania) amazonensis from the Amazon region. Phlebovirus coinfection with L. (L.) amazonensis in murine macrophages led to increased intracellular growth of L. (L.) amazonensis. Further studies with Icoaraci coinfection revealed the requirement of the PKR/IFN1 axis on the exacerbation of the parasite infection. L. (L.) amazonensis and Phlebovirus coinfection potentiated PKR activation and synergistically induced the expression of IFNβ and IL-10. Importantly, in vivo coinfection of C57BL/6 mice corroborated the in vitro data. The exacerbation effect of RNA virus on parasite infection may be specific because coinfection with dengue virus (DENV2) exerted the opposite effect on parasite load.
Altogether, our data suggest that coinfections with specific RNA viruses shared by vectors or reservoirs of Leishmania may enhance and sustain the activation of host cellular RNA sensors, resulting in aggravation of the parasite infection. The present work highlights new perspectives for the investigation of antiviral pathways as important modulators of protozoan infections.
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