Concentration In Culture Medium Research Articles

EFFECT OF THE CONCENTRATION OF THE PLANT GROWTH REGULATORS ON IN VITRO SHOOT GROWTH OF BLACKBERRY

Berry fruit species, such as Ribes nigrum, are highly esteemed in numerous countries for their biological and economic value, being regarded as important species of small fruits. Ribes nigrum, indigenous to northern Europe, and northern and central Asia. Conserving species like blackcurrant, as highlighted in the International Scientific reports, is crucial due to their potential health benefits and contributions to maintaining vascular health, especially in postmenopausal populations. The discovery that blackcurrant extract, rich in phytoestrogens, shows promise in preventing elastin degradation and pathological vascular remodeling underscores the importance of preserving this species. In Kazakhstan, the preservation of Ribes nigrum includes various factors, such as ecological significance, local economic value through agriculture, and conservation priorities since blackcurrant plays a significant role in the country's biodiversity and traditional uses. The potential of in vitro propagation for rapid mass production of valuable species is substantial. Our study addresses biodiversity conservation by employing this approach. Specifically, we aimed to investigate the impact of nutrient media and growth regulators on shoot multiplication of Ribes nigrum. Our findings reveal that the optimal concentration of plant growth regulators (PGR) significantly influences both the number and length of shoots. Notably, the highest multiplication rate for Ribes nigrum was achieved on MS with a combination of BAP at 0.5 μ/L and GA 0.5 μ/L. In conclusion, our study elucidates the effects of manipulating PGR concentrations in culture media on Ribes nigrum. We recommend optimizing these concentrations to enhance the growth of Ribes nigrum for potential reintroduction efforts and biodiversity conservation. The research is being conducted as part of program BR21882166, which focuses on the scientific and practical aspects of reproducing, conserving, and utilizing fruit and berry plants from the natural flora of Western, Eastern, Central, and Northern Kazakhstan to ensure food security. This program is financially supported by the Ministry of Science and Higher Education of the Republic of Kazakhstan for the period 2023-2025.

Serotonin-Salt-Stress Model-Induced Cell Growth via Promoting an Antioxidant System and Secondary Metabolites in Capsicum annuum Cell Suspension Culture.

Capsicum annuum contains potential therapeutic capsaicinoids, and various stress factors influence plant productivity. Serotonin is an indoleamine involved in signaling several stress response mechanisms in plants. However, the influence of serotonin on cell growth and the accumulation of secondary metabolites, mainly capsaicinoids production, is not yet clearly defined under salt stress. In this study, we optimized chili cell suspension cultures to maximize biomass, capsaicinoids, and phenolic compounds production using response surface methodology with two variables (serotonin and NaCl) of different concentrations in culture media supplemented with 2,4-dichlorophenoxyacetic acid and Kinetin. The results revealed a significant increase in biomass (14.3 g/L FW), capsaicin (0.93 μg/g FW), and dihydrocapsaicin content (0.32 μg/g FW) in chili cell suspension cultures compared with the control. Among all the phenolic compounds, chlorogenic acid was enhanced (17.4 μg/g FW), compared to control cultures. Serotonin exhibited stress mitigation effects and boosted antioxidant potential in chili suspension cultures. The present results illustrated that the optimized conditions can be used in scale-up studies of capsaicinoids production through the bioreactor.

P-190 Impact of low lactate culture media in embryo development and IVF outcomes

Abstract Study question Can low lactate culture media improve embryo utilization rate in artificial reproductive technology (ART) cycles? Summary answer Embryos cultured in low lactate media showed a significant improved blastocyst utilization rate compared to embryos cultured in standard culture media. What is known already Culture media plays a key role in the embryonic development in the IVF lab. Currently, different culture medias are commercially available, yet no consensus on the most suitable media composition has been reached. Recently, minimizing metabolic stress for the embryo to enhance implantation and pregnancy rates has been suggested. Research on the metabolism of mammalian preimplantation embryos revealed that excess lactate in the culture medium, in addition to pyruvate and glucose, can burden metabolic efficiency, as embryos naturally produce lactate during energy production. Study design, size, duration The prospective, monocentric study was designed and conducted at the Kinderwunsch Institut Schenk GmbH (Dobl, Austria). A total of 60 women with an average age of 32.5 undergoing ICSI (intracytoplasmic sperm injection) treatment were included. All of them had at least 6 mature (MII) oocytes at time of insemination. Sibling oocytes were cultured in two different culture medias and outcomes were compared. Patients undergoing preimplantation genetic testing were excluded. Participants/materials, setting, methods Mature (MII) oocytes of each patient (total 621 MII oocytes) were inseminated by ICSI and then equally split into two culture medias: standard (313 oocytes) and low lactate (308 oocytes) covered with mineral-oil overlay and incubated in time-lapse under the same conditions. A total of 428 embryos were obtained. Embryos were evaluated using morphokinetic criteria. The fertilization rate, embryo utilization rate, pregnancy rate and time to Blastocyst were analyzed. Main results and the role of chance No differences were observed in the number of fertilized oocytes (n = 211 and 217 in the standard and low lactate media, respectively). The overall embryo utilization rate was significantly different (p < 0.008) resulting in 52% in standard media and 64.3% using low lactate media. Interestingly, age stratification (18-30; 31-35; 36-41) revealed overall higher embryo utilization rates when using low lactate media in all age groups. Blastocyst utilization rate was significantly different (p < 0.007) 43,1% and 58,05% in the standard and low lactate media, respectively. Regarding the time to blastocyst, it was observed that in the standard culture media 80% of the blastocysts reached blastocyst stage on D + 5 whereas in the low lactate media 92,07% reached blastocyst stage on D + 5, which was also significantly different (p < 0.001). No differences in pregnancy and live birth rates were observed. Limitations, reasons for caution The number of patients may be seen as a study limitation. Results should be confirmed with a bigger sample size. Wider implications of the findings Low lactate media could help improve IVF outcomes due to higher embryo utilization rate leading to better cumulative pregnancy rates. Excessive lactate concentration in culture media could negatively influence embryo metabolism. Trial registration number not applicable

Formation of the pyruvoyl-dependent proline reductase Prd from Clostridioides difficile requires the maturation enzyme PrdH.

Stickland fermentation, the coupled oxidation and reduction of amino acid pairs, is a major pathway for obtaining energy in the nosocomial bacterium Clostridioides difficile. D-proline is the preferred substrate for the reductive path, making it not only a key component of the general metabolism but also impacting on the expression of the clostridial toxins TcdA and TcdB. D-proline reduction is catalyzed by the proline reductase Prd, which belongs to the pyruvoyl-dependent enzymes. These enzymes are translated as inactive proenzymes and require subsequent processing to install the covalently bound pyruvate. Whereas pyruvoyl formation by intramolecular serinolysis has been studied in unrelated enzymes, details about pyruvoyl generation by cysteinolysis as in Prd are lacking. Here, we show that Prd maturation requires a small dimeric protein that we have named PrdH. PrdH (CD630_32430) is co-encoded with the PrdA and PrdB subunits of Prd and also found in species producing similar reductases. By producing stable variants of PrdA and PrdB, we demonstrate that PrdH-mediated cleavage and pyruvoyl formation in the PrdA subunit requires PrdB, which can be harnessed to produce active recombinant Prd for subsequent analyses. We further created PrdA- and PrdH-mutants to get insight into the interaction of the components and into the processing reaction itself. Finally, we show that deletion of prdH renders C. difficile insensitive to proline concentrations in culture media, suggesting that this processing factor is essential for proline utilization. Due to the link between Stickland fermentation and pathogenesis, we suggest PrdH may be an attractive target for drug development.

Comparison of NK cell count between 1% and 10% FBS culture media concentration by FACS analysis

Recently, there has been a growing research interest in the application of immunotherapy for cancer treatment. Various approaches have been developed, and one of these methods involves utilizing Natural Killer (NK) cells of the immune system to specifically target cancer cells. Research on this method necessitates the mass and stable production of NK cells through culturing. For NK cell culture, Fetal Bovine Serum (FBS) is widely used in the industry at concentrations ranging from 10% to 20%. In an effort to explore a more efficient culturing technique, this study aims to investigate the effect of 1% FBS on cell viability of NK cells in cell culture. Because NK cells are usually defined by cell surface molecules, the efficacy of using a lower FBS concentration was quantitatively measured using Fluorescence-Activated Cell Sorting (FACS). The cell counts of cells cultured in 1% FBS was compared to those of cells cultured in 10% FBS. The results showed that no significant difference was found between cell count of cells cultured in 1% and 10% FBS. Thus, 1% FBS concentration culturing method was not found to be inferior. Further research is needed on the effect of replicating cancer microenvironment on NK cell viability.

Metabolic Profiling of SH-SY5Y and Neuro2A Cells in Relation to Fetal Calf Serum (FCS) Concentration in Culture Media.

The neuroblastoma cell lines SH-SY5Y and Neuro2A are commonly utilized models in neurobiological research. DMEM supplemented with different nutrients and 5-10% Fetal Calf Serum (FCS) is typically used for culturing these cell lines. During special treatments, a reduced FCS content is often deployed to reduce cellular proliferation or the content of bioactive compounds. The impact of the reduction of FCS in culture media on the metabolic profile of SH-SY5Y and Neuro2A cells is currently unknown. Using an Amplex Red Assay, this study showed that the consumption of L-glutamine decreased after FCS reduction. Glucose and pyruvate consumption increased in both cell lines after the reduction of FCS. Thus, lactate production also increased with reduced FCS concentration. The reduction of FCS in the cell culture medium resulted in a reduced aerobic ATP production for SH-SY5Y cells and a complete shut down of aerobic ATP production for Neuro2A cells, measured using the Seahorse XF Real-Time ATP Rate Assay. Utilizing the Seahorse XF Glutamine Oxidation Stress Test, Neuro2A cells showed an increased utilization of L-glutamine oxidation after reduction of FCS. These results indicate that changes in FCS concentration in culture media have an impact on the different energy production strategies of SH-SY5Y and Neuro2A cells which must be considered when planning special treatments.

Soyghurt Potentially Controls the Level of sFlt1 and PLGF in Preeclampsia Maternal Serum-Induced Placental Trophoblast Cell in vitro.

To observe the effect of soya yoghurt (Soyghurt), which is high in flavonoid substance, on the expression of preeclampsia biomarkers (sFLT-1 and PLGF) on preeclampsia serum-induced trophoblast primary cell culture isolated from placental tissue. The trophoblast primary culture was induced by preeclampsia serum (10%). The Soyghurt treatment was performed with 2.5%, 5%, and 7.5% Soyghurt supernatant concentrations in culture media. The expression of preeclampsia markers, sFLT-1 and PLGF, were evaluated using ELISA. Expression of sFLT-1 on preeclampsia-induced cell culture treated with Soyghurt was significantly lowered compared to the untreated group (p<0.01). However, no significant difference was observed in the PLGF levels of all groups induced by preeclampsia serum (p>0.05). This study demonstrates the potential effect of Soyghurt's in balancing preeclampsia marker expression by inhibiting the expression of sFLT-1 in preeclampsia serum -induced trophoblast cells.

A fluorescent probe for selective detection of lysosomal β-hexosaminidase in live cells

Determining the activity of lysosomal β-hexosaminidase in cells is of great importance for understanding the roles that these enzymes play in pathophysiological events. Herein, we designed the new fluorescent probe, βGalNAc-Rhod-CM(NEt2), which consisted of a βGalNAc-linked rhodol unit serving as a β-hexosaminidase reactive fluorogenic moiety and a N,N′-diethylaminocoumarin (CM(NEt2)) group acting as a fluorescence marker for determining the degree of cell permeabilization. Treatment of βGalNAc-Rhod-CM(NEt2) with β-hexosaminidase promoted generation of Rhod-CM(NEt2), thereby leading to an increase in the intensity of fluorescence of Rhod. However, this probe did not respond to the functionally related glycosidase, O-GlcNAcase. The detection limit of βGalNAc-Rhod-CM(NEt2) for β-hexosaminidase was determined to be 0.52 nM, indicating that it has high sensitivity for this enzyme. Furthermore, the probe functioned as an excellent fluorogenic substrate for β-hexosaminidase with kcat and Km values of 17 sec−1 and 22 μM, respectively. The results of cell studies using βGalNAc-Rhod-CM(NEt2) showed that levels of β-hexosaminidase activity in cells can be determined by measuring the intensity of fluorescence arising from Rhod and that the intensity of fluorescence of CM(NEt2) can be employed to determine the degree of cell permeabilization of the probe. Utilizing the new probe, we assessed β-hexosaminidase activities in several types of cells and evaluated the effect of glucose concentrations in culture media on the activity of this enzyme.

Changes in Lactic Acid Concentrations in Culture Media and its Implications in the Inhibition of Fungal Development

In the following research, a novel antifungal culture method was discussed with the aim of developing bacterial culture media in which fungal growth is inhibited and bacterial growth is stimulated. This technique is based on the use of lactic acid as an antifungal agent, due to its capacity to neutralise the electrochemical gradient of fungal cell membranes, thus inhibiting their development. Based on this principle, the Minimum Inhibitory Concentration (MIC) of lactic acid in the culture media was Investigated by changing its concentrations. After the preparation of the solutions, microbiological seeding was carried out in the different experimental groups. Data were collected using a quadrat sampling technique for subsequent statistical analysis using the T-test coefficient. As a result, the minimum inhibitory concentration of lactic acid was found to be 20%, where fungal growth was inhibited, and bacterial growth was stimulated.

Regulatory-systemic approach in Aspergillus niger for bioleaching improvement by controlling precipitation.

In the context of e-waste recycling by fungal bioleaching, nickel and cobalt precipitate as toxic metals by oxalic acid, whereas organic acids, such as citric, act as a high-performance chelating agent in dissolving these metals. Oxalic acid elimination requires an excess and uneconomical carbon source concentration in culture media. To resolve this issue, a novel and straightforward systems metabolic engineering method was devised to switch metabolic flux from oxalic acid to citric acid. In this technique, the genome-scale metabolic model of Aspergillus niger was applied to predicting flux variability and key reactions through the calculation of multiple optimal solutions for cellular regulation. Accordingly, BRENDA regulators and a novel molecular docking-oriented approach were defined a regulatory medium for this end. Then, ligands were evaluated in fungal culture to assess their impact on organic acid production for bioleaching of copper and nickel from waste telecommunication printed circuit boards. The protein structure of oxaloacetate hydrolase was modeled based on homology modeling for molecular docking. Metformin, glutathione, and sodium fluoride were found to be effective as inhibitors of oxalic acid production, enabling the production of 8100 ppm citric acid by controlling cellular metabolism. Indirect bioleaching demonstrated that nickel did not precipitate, and the bioleaching efficiency of copper and nickel increased from 40% and 24% to 61% and 100%, respectively. Bioleaching efficiency was evaluated qualitatively by FE-SEM, EDX, mapping, and XRD analysis. KEY POINTS: • A regulatory-systemic procedure for controlling cellular metabolism was introduced • Metformin inhibited oxalic acid, leading to 8100 ppm citric acid production • Bioleaching of copper and nickel in TPCBs improved by 21% and 76.

Macrophage phenotype impacts in vitro equine intrasynovial deep digital flexor tenocyte matrix metalloproteinase gene expression and secretion.

To investigate matrix metalloproteinase (MMP) and their inhibitors tissue inhibitor matrix metalloproteinase (TIMP) gene expression and secretion during equine deep digital flexor tendon (DDFT) tenocyte and macrophage (undifferentiated, proinflammatory, and regulatory) co-culture. Third passage DDF tenocytes and donor-matched macrophages differentiated from peripheral blood CD14+ monocytes from 5 healthy horses ages 9-11 years, euthanized for reasons unrelated to musculoskeletal conditions. Passage 3 DDT tenocyte aggregate cultures were co-cultured with undifferentiated (control), proinflammatory (granulocyte-macrophage colony-stimulating factor; GM-CSF pretreated and lipopolysaccharide + interferon gamma-primed; LPS+IFN-γ) or regulatory (interleukin-4 and interleukin-10-primed; IL-4 + IL-10) macrophages in direct and transwell co-cultures for 72 hours. MMP-1, -2, -3, -9, -13, and TIMP -1, -2 mRNA were measured via real-time Polymerase Chain Reaction (rtPCR). Co-culture media MMP -3, -9, and TIMP -1, -2 concentrations were quantified via ELISA. Direct co-culture of DDF tenocytes with proinflammatory macrophages for 72 hours increased MMP-1, -3, and -13 mRNA levels whereas, MMP-9 mRNA levels decreased. Direct and transwell co-culture with proinflammatory and regulatory macrophages resulted in increased MMP-3 and decreased MMP-9 media concentrations. While direct co-culture with regulatory macrophages significantly increased TIMP-1 mRNA, overall, TIMP mRNA and culture media concentrations were largely unchanged. Cell-to-cell contact between DDF tenocytes and macrophages is not essential to induce MMP gene expression and secretion. Co-culture systems offer a viable in vitro platform to screen and evaluate immunomodulatory properties of therapies aimed at improving equine intrasynovial tendon healing.

Research on the growth ability of Saccharomyces cerevisiae MN2 in the medium containing different concentrations of NaCl and NH\(_{4}^{+}\) for applying in aquaculture

Salinity is one of the factors affecting the growth and activity of microorganisms. The study of the salinity effect can indicate the range of microorganism applications. This study evaluated the effects of NaCl concentration on growth, amino acid and NH4+ absorption of yeast strain Saccharomyces cerevisia MN2. The results showed that, within NaCl concentration from 0 to 3%, the negative effects on the growth of MN2 strain were not observed. The cell density on all culture media reached 108 CFU/ml after 24 hours and maintained until 48 hours.The strain NM2 was able to absorb amino acids at a high rate, which was indicated by the decrease of the amino acid concentration in the culture media after 48 hours (from 20.8 to 37.5%) in comparison with the control. In addition, the strain NM2 was also able to decrease NH4+ concentration in culture media, reducing from 17 to 25.4% in comparison with the control after 48 hours. Strain MN2 is capable of using ammonium as a nitrogen source. When the ammonium concentration fluctuated in the range of 37,327 - 75,433 mg/l, it did not affect the growth of MN2 much. In conclusion, the strain of S. cerevisia MN2 is tolerance to the salinity up to 3%, and suitable for the aquaculture wastewater treatment process.

MP20-07 NPTX1 IS A NOVEL SERUM BIOMARKER INDUCED BY ARPI TREATED PROSTATE CANCER AND ENRICHED IN LINEAGE PLASTICITY WITH BRN2 HIGH

You have accessJournal of UrologyCME1 Apr 2023MP20-07 NPTX1 IS A NOVEL SERUM BIOMARKER INDUCED BY ARPI TREATED PROSTATE CANCER AND ENRICHED IN LINEAGE PLASTICITY WITH BRN2 HIGH Takeshi Namekawa, Joshua Scurll, Maxim Kobelev, Dwaipayan Ganguli, Olena Sivak, Martin Gleave, Vancouver, Canada, Eva Corey, Colm Morrissey, and Amina Zoubeidi Takeshi NamekawaTakeshi Namekawa More articles by this author , Joshua ScurllJoshua Scurll More articles by this author , Maxim KobelevMaxim Kobelev More articles by this author , Dwaipayan GanguliDwaipayan Ganguli More articles by this author , Olena SivakOlena Sivak More articles by this author , Martin GleaveMartin Gleave More articles by this author , Vancouver Vancouver More articles by this author , Canada Canada More articles by this author , Eva CoreyEva Corey More articles by this author , Colm MorrisseyColm Morrissey More articles by this author , and Amina ZoubeidiAmina Zoubeidi More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003245.07AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Androgen receptor (AR) pathway inhibition is the first-line therapy for metastatic prostate cancer (PCa). However, tumors invariably develop resistance to ARPI. We previously identified a transcription factor Brain-2 (BRN2), encoded by the gene POU3F2, as a master driver of NEPC which is an aggressive form of AR-independent phenotypes treatment resistant PCa. In this study, we identified a surrogate biomarker of BRN2 activity to monitor the progression of treatment resistant PCa. METHODS: To identify BRN2 serum marker, we analyzed BRN2 ChIPseq data from 2 NEPC and 2 neural-type lung squamous cell carcinoma (nLUSC) cell lines with a focus on shared BRN2 peaks. Annotated peaks from enhancers (EnhancerAtlas) and promoters (genes within 20kb) were selected for secreted proteins and were ranked by correlation with BRN2 expression in PCa patients. Validation of protein expression was performed by western blot. R-PLEX Assay kit was used to measure concentrations in culture media from NCI-H660 cells and in serum from mice with NCI-H660 xenograft or PDX tumors and from human PCa patients. RESULTS: We identified 51 candidate serum biomarkers for BRN2 activity (or expression?) using the methods described above. These biomarkers were ranked based on their expression and correlation with BRN2 expression in PCa patient samples across four public data sets. NPTX1 was identified as the top biomarker of BRN2 expression. We further validated association of NPTX1 and BRN2 expression by BRN2 overexpression in MR42D and 22Rv1, indicating that BRN2 positively regulates NPTX1 expression. Next, we showed that NPTX1 is present in culture media from NCI-H660 cells, and that NPTX1 serum concentration positively correlated with tumor volume in mice with NCI-H660 xenografts and with NPTX1 mRNA expression in LuCaP PDX models. Importantly, evaluation of patients’ samples showed that NPTX1 levels were high in 26/35 CRPC serum samples vs levels in serum of healthy volunteers, and that the NPTX1 serum levels were highest in 4/6 patients with high POU3F2 tumor expression. CONCLUSIONS: We identified and validated NPTX1 as a serum marker that can be readily detected in serum from NEPC patients with BRN2high tumours. Source of Funding: U.S. Army Medical Research and Development Command(W81XWH1810689) © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e277 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Takeshi Namekawa More articles by this author Joshua Scurll More articles by this author Maxim Kobelev More articles by this author Dwaipayan Ganguli More articles by this author Olena Sivak More articles by this author Martin Gleave More articles by this author Vancouver More articles by this author Canada More articles by this author Eva Corey More articles by this author Colm Morrissey More articles by this author Amina Zoubeidi More articles by this author Expand All Advertisement PDF downloadLoading ...

Interleukin-1β and methylprednisolone acetate demonstrate differential effects on equine deep digital flexor tendon and navicular bone fibrocartilage cells in vitro

To investigate the effects of interleukin-1β (IL-1β) and methylprednisolone acetate (MPA) on equine intrabursal deep digital flexor tendon (DDFT) and navicular bone fibrocartilage (NBF) cells in vitro. Third passage DDFT and NBF cells from 5 healthy donor horses ages 11-17 years euthanized for reasons unrelated to musculoskeletal conditions. Aggregate cultures were incubated with culture medium alone (control), 10 ng/mL IL-1β, 10 ng/mL IL-1β + 0.05 mg/mL MPA, or 10 ng/mL IL-1β + 0.5 mg/mL MPA for 24 hours. Extracellular matrix (ECM) gene expressions were assessed via real-time polymerase chain reaction (rtPCR). Culture media matrix metalloproteinase (MMP) -3 and -13 concentrations were quantified via ELISA. Total glycosaminoglycan (GAG) content in the cell pellets and culture media was also assessed. IL-1β and IL-1β combined with MPA significantly downregulated ECM gene expression to a greater extent in NBF cells compared with DDFT cells. IL-1β and IL-1β combined with MPA significantly upregulated MMP-3 culture media concentrations in DDFT cells only, and MMP-13 culture media concentrations to a greater extent in NBF cells compared with DDFT cells. NBF cells were more susceptible to IL-1β and MPA-mediated ECM gene expression downregulation in vitro. These results serve as a first step for future work to determine intrabursal corticosteroid regimens that limits or resolve the inflammation as well as take into consideration NBF cell biosynthesis in horses with navicular disease, for which currently no information exists.

Letrozole increases preantral follicle growth and decreases estradiol production without impairing follicle survival

BackgroundLetrozole has been reported to be effective in treating anovulation, preventing ovarian hyperstimulation syndrome (OHSS), and retrieving oocytes in breast cancer patients. However, the role and mechanism of letrozole in follicular development remain unclear.ResultsWe treated mouse preantral follicles with various treatments; we found no significant difference in follicle survival rates in the letrozole (LET) group compared with the control group, but the average diameter of follicles in the LET group tended to be larger (CTRL vs. LET 30, p = 0.064; CTRL vs. LET 100, p = 0.025). The estradiol concentrations in culture media of the LET group were significantly lower than those observed in the control group (CTRL vs. LET 30, p = 0.038; CTRL vs. LET 100, p = 0.025). We further found a marked increase in follicle-stimulating hormone receptor (FSHR) gene expression in response to letrozole treatment (CTRL vs. LET 30, p = 0.075; CTRL vs. LET 100, p = 0.034). This result suggested that increased FSHR expression promotes follicle development. Letrozole inhibited aromatase activity, but the effect was limited. Letrozole did not significantly reduce vascular endothelial growth factor (VEGF) gene expression.ConclusionsLetrozole may promote follicle development by increasing the expression of FSHR. Letrozole may be useful for fertility preservation of patients with estrogen-dependent cancers such as breast cancer and various other cancers. Whether letrozole has a direct effect in reducing OHSS requires further investigation.

Hydrogen peroxide pretreatment assisted phytoremediation of sodium dodecyl sulfate by Juncus acutus L

BackgroundSodium Dodecyl Sulfate (SDS) an anionic surfactant pollutant has emerged as a serious hazard to the aquatic and terrestrial environment. Due to physical and chemical methodological difficulties for SDS removal, phytoremediation techniques are efficient alternative strategies to tackle such adversities. Juncus acutus L. (J. acutus) is a pioneer wetland species that has been recently exploited for phytoremediation purposes. To our knowledge, the role of exogenous hydrogen peroxide (H2O2), in improving the phytoextraction of SDS has not been examined yet. In this study, pretreatment foliar spray of H2O2 (15 mM) combined with two levels of SDS (50 and 100 ppm) in water culture was evaluated to remove SDS contamination and add value to the phytoremediation process.ResultsThe outcomes revealed that J. acutus has considerable translocation and bioaccumulation abilities for SDS and can be utilized as an appropriate hyperaccumulator in SDS-contaminated sites. However, the involvement of H2O2 extended phytoremediation capacity and successive removal of SDS. H2O2 significantly assisted in increasing SDS remediation via more accumulation in J. acutus tissues by 29.9 and 112.4% and decreasing SDS concentration in culture media by 33.3 and 27.3% at 50 and 100 ppm SDS, respectively. Bioaccumulation factor (BCF) increased by 13.8 and 13.2%, while translocation factor (TCF) positively maximized by 82.4 and 76.2% by H2O2 application at 50 and 100 ppm SDS, respectively. H2O2 pretreatment could drive the decline in biochemical attributes in SDS-affected plants by modulating stress tolerance indices, pigments, water relations, proline content, enzymatic activities, and further, reduced oxidative stress in terms of electrolyte leakage, cellular H2O2, malondialdehyde (MDA) accumulation.ConclusionsH2O2 could play a potential role in maximizing phytoremediation capacity of SDS by J. acutus in polluted sites.

Investigating the effects of 5.5 mmoL vs 25 mmoL glucose concentration in culture media on LHCN-M2 cell viability, proliferation, metabolism and differentiation

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Microbial Astaxanthin Production from Agro-Industrial Wastes—Raw Materials, Processes, and Quality

The antioxidant and food pigment astaxanthin (AX) can be produced by several microorganisms, in auto- or heterotrophic conditions. Regardless of the organism, AX concentrations in culture media are low, typically about 10–40 mg/L. Therefore, large amounts of nutrients and water are necessary to prepare culture media. Using low-cost substrates such as agro-industrial solid and liquid wastes is desirable for cost reduction. This opens up the opportunity of coupling AX production to other existing processes, taking advantage of available residues or co-products in a biorefinery approach. Indeed, the scientific literature shows that many attempts are being made to produce AX from residues. However, this brings challenges regarding raw material variability, process conditions, product titers, and downstream processing. This text overviews nutritional requirements and suitable culture media for producing AX-rich biomass: production and productivity ranges, residue pretreatment, and how the selected microorganism and culture media combinations affect further biomass production and quality. State-of-the-art technology indicates that, while H. pluvialis will remain an important source of AX, X. dendrorhous may be used in novel processes using residues.

Nanoscale Detonation Carbon Demonstrates Biosafety in Human Cell Culture

The production method of nanoscale detonation carbon (NDC) has recently been developed at Lavrentyev Institute of Hydrodynamics SB RAS. This method uses the reaction of acetylene with oxygen conducted in the detonation mode in fuel-rich acetylene–oxygen mixtures. The morphology and structural features of the NDC particles can be varied by changing the concentration of oxygen in the gaseous mixtures. The particles of NDC can serve as reinforcements in metal matrix composites and additives imparting electrical conductivity to polymer matrix composites. Before NDC can be considered for industrial applications, it is necessary to address the related biological safety concerns. The present work was aimed at determining the cytotoxicity of NDC. The NDC powders with two morphologies (obtained using different acetylene/oxygen ratios) were tested on HEK293A human cells. The NDC powder was added to the culture medium in concentrations ranging from 10 ng/mL to 400 μg/mL. The cell viability was determined by a colorimetric EZ4U test and a real-time cell analyzer xCELLigence. None of the NDC samples showed a cytotoxic effect. The results of this study allow us to recommend NDC as a safe and useful product for the development of advanced carbon-based and composite materials.

A comparison of the lowest effective concentration in culture media for detection of chromosomal damage in vitro and in blood or plasma for detection of micronuclei in vivo.

It is often assumed that genotoxic substances will be detected more easily by using in vitro rather than in vivo genotoxicity tests since higher concentrations, more cytotoxicity and static exposures can be achieved. However, there is a paucity of data demonstrating whether genotoxic substances are detected at lower concentrations in cell culture in vitro than can be reached in the blood of animals treated in vivo. To investigate this issue, we compared the lowest concentration required for induction of chromosomal damage in vitro (lowest observed effective concentration, or LOEC) with the concentration of the test substance in blood at the lowest dose required for biologically relevant induction of micronuclei in vivo (lowest observed effective dose, or LOED). In total, 83 substances were found for which the LOED could be identified or estimated, where concentrations in blood and micronucleus data were available via the same route of administration in the same species, and in vitro chromosomal damage data were available. 39.8 % of substances were positive in vivo at blood concentrations that were lower than the LOEC in vitro, 22.9 % were positive at similar concentrations, and 37.3 % of substances were positive in vivo at higher concentrations. Distribution analysis showed a very wide scatter of >6 orders of magnitude across these 3 categories. When mode of action was evaluated, the distribution of clastogens and aneugens across the 3 categories was very similar. Thus, the ability to detect induction of micronuclei in bone marrow in vivo regardless of the mechanism for micronucleus induction, is clearly not solely determined by the concentration of test substance which induced chromosomal damage in vitro.