Conventional Medium Research Articles

Comparative evaluation of acrylamide and polycyclic aromatic hydrocarbons contents in Robusta coffee beans roasted by hot air and superheated steam

Although hot air (HA) is a conventional roasting medium for coffee beans, HA roasting is known to result in possible formation of toxic compounds, including acrolein, acrylamide and polycyclic aromatic hydrocarbons (PAHs). Superheated steam (SHS) roasting is therefore proposed as an alternative means to alleviate the formation of these toxic compounds in roasted coffee beans. Robusta coffee beans were roasted either with HA or SHS in a fluidized bed roaster at 210–250 °C until the bean color reached the targeted roast levels. The contents of acrolein, acrylamide and 16 PAHs in the roasted beans were determined; only acrylamide and 5 PAHs were nevertheless found. SHS roasting interestingly resulted in lower acrylamide contents in dark-roasted beans; similar trend was noted in the beans medium-roasted at 250 °C. The contents of three-ring PAHs, namely fluorene, phenanthrene and anthracene, in dark-roasted beans were significantly lower upon SHS roasting at 250 °C.

In Vitro Culture of Chicken Circulating and Gonadal Primordial Germ Cells on a Somatic Feeder Layer of Avian Origin.

Simple SummaryChicken primordial germ cells are specialized cells that are formed outside the developing embryo, from where they migrate into the gonad, and give rise to the gametes. There are two types of those cells: Circulating blood primordial germ cells and gonadal primordial germ cells. They can be isolated only in low numbers from the bloodstream or gonads of donor embryos. Hence, efficient in vitro cultivation systems are required to increase their quantity through proliferation. Here, we provide a single culture system that can be used to cultivate both cell types. We also present a novel, easy-to-train, and non-invasive method to identify live primordial germ cells in the culture. In this analysis, chicken primordial germ cells obtained from embryonic blood or gonadal regions were cultured in vitro on feeder cell layers derived from embryonic chick cells. The use of the chicken origin feeder layer allowed reducing the xenogenic animal factors in the culture. We demonstrated a feasible and cost-efficient technique to routinely assess the cultivated primordial germ cells on the basis of their morphological characteristics and using the optical features of cells in darkfield illumination. This method is especially useful to distinguish primordial germ cells during co-cultivation with other cell types.The present study had two aims: (1) To develop a culture system that imitates a normal physiological environment of primordial germ cells (PGCs). There are two types of PGCs in chicken: Circulating blood (cPGCs) and gonadal (gPGCs). The culture condition must support the proliferation of both cPGCs and gPGCs, without affecting their migratory properties and must be deprived of xenobiotic factors, and (2) to propose an easy-to-train, nonlabeling optical technique for the routine identification of live PGCs. To address the first aim, early chicken embryo’s feeder cells were examined instead of using feeder cells from mammalian species. The KAv-1 medium at pH 8.0 with the addition of bFGF (basic fibroblast growth factor) was used instead of a conventional culture medium (pH approximately 7.2). Both cPGCs and gPGCs proliferated in vitro and retained their migratory ability after 2 weeks of culture. The cultivated cPGCs and gPGCs colonized the right and/or left gonads of the recipient male and female embryos. To address the second aim, we demonstrated a simple and rapid method to identify live PGCs as bright cells under darkfield illumination. The PGCs rich in lipid droplets in their cytoplasm highly contrasted with the co-cultured feeder layer and other cell populations in the culture.

Exploiting the use of agro-industrial residues from fruit and vegetables as alternative microalgae culture medium

There is a need for searching new microalgae species, and the most suitable strategy to increase the cost-effectiveness of a microalgae culture system is to use resources of low costs, such as residues. This study aimed to evaluate the cultivation of microalgae isolated from the Brazilian Northeast region (Lagerheimia longiseta, Monoraphidium contortum, and Scenedesmus quadricauda) in an alternative medium of low cost (biocompost of discarded fruits and vegetables) with a view to possible applications in the food industry. Microalgae cultivated in the conventional synthetic medium was used as control. The cultivation of microalgae in the alternative medium allowed suitable cell growth, and improved the antioxidant activity and the levels of monounsaturated fatty acid and polyunsaturated fatty acid compared to the synthetic medium. The cultivation of S. quadricauda and L. longiseta species in the alternative medium resulted in increased protein content and/or total phenolic content, and improved health indices (lower levels of atherogenic, thrombogenic, and hypercholesterolemic saturated fatty acids indices, and higher levels of desired fatty acids index) compared to cultivation in synthetic medium. The cultivation of M. contortum in the alternative medium contributed to the production of higher lipid content, mainly saturated fatty acid (palmitic acid), which contributed negatively to the health indices. This study proved that S. quadricauda and L. longiseta microalga species from freshwaters have significant potential for distinct applications in functional food industries, and the biocompost of discarded fruits and vegetables is a suitable medium for microalgae cultivation.

Meios alternativos constituídos por subproduto do cultivo de microrganismos fotossintetizantes para o crescimento de Lactobacillus acidophilus e produção de β-Galactosidase

Bactérias ácido-láticas como Lactobacillus acidophilus possuem grande potencial de mercado devido aos seus efeitos probióticos e no processamento de produtos lácteos. Essas bactérias também sintetizam enzimas de interesse industrial, como a β-galactosidase (lactase). No entanto, a produção em larga escala desse produto é onerosa devido à formulação do meio de cultivo, levando a busca por alternativas para substituição total ou parcial. O líquido metabólico (LM) de microrganismos fotossintetizantes pode ser uma opção devido à quantidade de vitaminas, açúcares e proteínas liberadas no meio. A reutilização do LM é interessante para a produção comercial de L. acidophilus por reduzir os custos e prevenir a contaminação ambiental. O objetivo deste estudo foi avaliar o crescimento de L. acidophilus e a produção de β-galactosidase em meios de cultura contendo LM de microrganismos fotossintetizantes. L. acidophilus foi cultivada em meio convencional contendo diferentes concentrações de líquido metabólico de quatro microrganismos fotossintetizantes (Artrhospira platensis, Dunaliella tertiolecta, Chlorella vulgaris e Scenedesmus sp.) durante 72 horas, a 37 ºC. A cada 24h, foram avaliados o crescimento celular e a produção da enzima β-galactosidase. No geral, os maiores crescimentos observados foram nos meios contendo 25% e 50% de líquido metabólico após 24h de cultivo. A melhor condição observada foi com 50% do líquido metabólico de C. vulgaris, obtendo-se mais de 60% e 90% do crescimento celular e da produção de β-galactosidase, respectivamente, quando comparado ao meio convencional, mostrando ser um resíduo líquido alternativo na substituição parcial do meio pelo crescimento de L. acidophilus e produção de β-galactosidase.

Development of a low-cost culture medium for the rapid production of plant growth-promoting Rhodopseudomonas palustris strain PS3.

Rhodopseudomonas palustris PS3 is one of the purple phototrophic non-sulfur bacteria (PNSB), which have plant growth-promoting effects on various plants. To expand the scale of PS3 fermentation in a time- and cost-effective fashion, the purpose of this work was to evaluate the use of low-cost materials as culture media and to optimize the culture conditions via response surface methodology. Corn steep liquor (CSL) and molasses were identified as potential materials to replace the nitrogen and carbon sources, respectively, in the conventional growth medium. The optimum culture conditions identified through central composite design were CSL, 39.41 mL/L; molasses, 32.35 g/L; temperature, 37.9°C; pH, 7.0; and DO 30%. Under the optimized conditions, the biomass yield reached 2.18 ± 0.01 g/L at 24 hours, which was 7.8-fold higher than that under the original medium (0.28 ± 0.01 g/L). The correlation between the predicted and experimental values of the model was over 98%, which verified the validity of the response models. Furthermore, we verified the effectiveness of the R. palustris PS3 inoculant grown under the newly developed culture conditions for plant growth promotion. This study provides a potential strategy for improving the fermentation of R. palustris PS3 in low-cost media for large-scale industrial production.

Corn Wastes and Peanut Shell as Growing Media for Production of Red Radish Plants in Soilless System

ABSTRACT Peat is considered the conventional growing medium in most soilless culture systems. The high cost of peat and the urgent need of agricultural wastes recycling encouraged the scientists and soilless culture users to search for an alternative growth medium where optimal growth conditions are achieved and help in the safe disposal of wastes. In the current study, peanut shell (PS) and corn wastes (CW) were used as growing media in comparison to peat moss (PM). The tested organic wastes and peat moss were examined with sand at three mixing ratios (1:1 “M1”, 1:2 “M2” and 1:3 “M3” raw material: washed sand, respectively). Red radish (Raphanus raphanistrum subsp. sativus) plant were cultivated in 5 kg pots filled with the instigated growing media. Most of the recoded growth parameters were found in PS and PM growing media, while the lowest ones were found in CW. PS medium contained available N and P higher by 141 and 29% above the PM media. Although the peat moss gave the highest values in the measured growth characteristics, its high price decreased the net profit. The highest net profit value was obtained from PSM2 followed by PSM1 and PSM3. According to the obtained results, the characteristics of the growth media derived from peanut shell qualify them for use in the production of red radish in soilless culture systems.

GM-CSF (granulocyte macrophage colony-stimulating factor) supplementation in culture media for women undergoing assisted reproduction.

GM-CSF (granulocyte macrophage colony-stimulating factor) is a growth factor that is used to supplement culture media in an effort to improve clinical outcomes for those undergoing assisted reproduction. It is worth noting that the use of GM-CSF-supplemented culture media often adds a further cost to the price of an in vitro fertilisation (IVF) cycle. The purpose of this review was to assess the available evidence from randomised controlled trials (RCTs) on the effectiveness and safety of GM-CSF-supplemented culture media. To assess the effectiveness and safety of GM-CSF-supplemented human embryo culture media versus culture media not supplemented with GM-CSF, in women or couples undergoing assisted reproduction. We used standard methodology recommended by Cochrane. We searched the Cochrane Gynaecology and Fertility Group Trials Register, CENTRAL, MEDLINE, Embase, CINAHL, LILACS, DARE, OpenGrey, PubMed, Google Scholar, and two trials registers on 15 October 2019, checked references of relevant papers and communicated with experts in the field. We included RCTs comparing GM-CSF (including G-CSF (granulocyte colony-stimulating factor))-supplemented embryo culture media versus any other non-GM-CSF-supplemented embryo culture media (control) in women undergoing assisted reproduction. We used standard methodological procedures recommended by Cochrane. The primary review outcomes were live birth and miscarriage rate. The secondary outcomes were clinical pregnancy, multiple gestation, preterm birth, birth defects, aneuploidy, and stillbirth rates. We assessed the quality of the evidence using GRADE methodology. We undertook one comparison, GM-CSF-supplemented culture media versus culture media not supplemented with GM-CSF, for those undergoing assisted reproduction. We included five studies, the data for three of which (1532 participants) were meta-analysed. We are uncertain whether GM-CSF-supplemented culture media makes any difference to the live-birth rate when compared to using conventional culture media not supplemented with GM-CSF (odds ratio (OR) 1.19, 95% confidence interval (CI) 0.93 to 1.52, 2 RCTs, N = 1432, I2 = 69%, low-quality evidence). The evidence suggests that if the rate of live birth associated with conventional culture media not supplemented with GM-CSF was 22%, the rate with the use of GM-CSF-supplemented culture media would be between 21% and 30%. We are uncertain whether GM-CSF-supplemented culture media makes any difference to the miscarriage rate when compared to using conventional culture media not supplemented with GM-CSF (OR 0.75, 95% CI 0.41 to 1.36, 2 RCTs, N = 1432, I2 = 0%, low-quality evidence). This evidence suggests that if the miscarriage rate associated with conventional culture media not supplemented with GM-CSF was 4%, the rate with the use of GM-CSF-supplemented culture media would be between 2% and 5%. Furthermore, we are uncertain whether GM-CSF-supplemented culture media makes any difference to the following outcomes: clinical pregnancy (OR 1.16, 95% CI 0.93 to 1.45, 3 RCTs, N = 1532 women, I2 = 67%, low-quality evidence); multiple gestation (OR 1.24, 95% CI 0.73 to 2.10, 2 RCTs, N = 1432, I2 = 35%, very low-quality evidence); preterm birth (OR 1.20, 95% CI 0.70 to 2.04, 2 RCTs, N = 1432, I2 = 76%, very low-quality evidence); birth defects (OR 1.33, 95% CI 0.59 to 3.01, I2 = 0%, 2 RCTs, N = 1432, low-quality evidence); and aneuploidy (OR 0.34, 95% CI 0.03 to 3.26, I2 = 0%, 2 RCTs, N = 1432, low-quality evidence). We were unable to undertake analysis of stillbirth, as there were no events in either arm of the two studies that assessed this outcome. Due to the very low to low quality of the evidence, we cannot be certain whether GM-CSF is any more or less effective than culture media not supplemented with GM-CSF for clinical outcomes that reflect effectiveness and safety. It is important that independent information on the available evidence is made accessible to those considering using GM-CSF-supplemented culture media. The claims from marketing information that GM-CSF has a positive effect on pregnancy rates are not supported by the available evidence presented here; further well-designed, properly powered RCTs are needed to lend certainty to the evidence.

Induced Fetal Human Muscle Stem Cells with High Therapeutic Potential in a Mouse Muscular Dystrophy Model.

SummaryDuchenne muscular dystrophy (DMD) is a progressive and fatal muscle-wasting disease caused by DYSTROPHIN deficiency. Cell therapy using muscle stem cells (MuSCs) is a potential cure. Here, we report a differentiation method to generate fetal MuSCs from human induced pluripotent stem cells (iPSCs) by monitoring MYF5 expression. Gene expression profiling indicated that MYF5-positive cells in the late stage of differentiation have fetal MuSC characteristics, while MYF5-positive cells in the early stage of differentiation have early myogenic progenitor characteristics. Moreover, late-stage MYF5-positive cells demonstrated good muscle regeneration potential and produced DYSTROPHIN in vivo after transplantation into DMD model mice, resulting in muscle function recovery. The engrafted cells also generated PAX7-positive MuSC-like cells under the basal lamina of DYSTROPHIN-positive fibers. These findings suggest that MYF5-positive fetal MuSCs induced in the late stage of iPSC differentiation have cell therapy potential for DMD.

Growth response of the diatom Chaetoceros species to the elemental ratios of Deep Ocean waters

Phytoplankton production, succession and community composition is limited by nutrients, temperature and spectral properties. Elemental ratios serve as indicators for predicting productivity of phytoplankton including diatom. Deep ocean water (DOW) as a marine natural resource is rich in inorganic nutrients. In the present study, as part of the OTEC program, an initial attempt was made to characterize quantitatively the elemental ratios in DOW and relate to growth of the diatom, Chaetoceros sp. In addition, the production of Chaetoceros sp. in DOW was compared with a conventional nutrient enrichment medium, generally used for phytoplankton growth. To assess the productive ability of DOW, factorial experiments with Chaetoceros sp. were carried out with (i) unaltered DOW [100% DOW], (ii) admixtures [50:50] of DOW and surface seawater (SSW) [50% DOW], (iii) plain SSW, (iv) SSW enriched with nutrients and (v) DOW enriched with nutrients. Results of admixtures of seawater starting from 0% SSW to 100% DOW revealed optimum Redfield ratios at 50% dilution. Results on growth of Chaetoceros sp. in 100% DOW showed a sudden blooming on the 2nd and 3rd day. Chlorophyll ‘a’ values of 70 mg m−3 were obtained with DOW and SSW enriched with f/2 medium. Similarly with 50% DOW same chlorophyll ‘a' concentrations was reached on the 4th 5th day and declined on the 6th day. Results of the present study demonstrate that 100% and 50% dilutions of DOW could be used for intensive culture of Chaetoceros sp. and that biomass production could be achieved within 3 days compared to incubations which use a conventional medium.

Transfer of healthy fibroblast-derived mitochondria to HeLa ρ0 and SAS ρ0 cells recovers the proliferation capabilities of these cancer cells under conventional culture medium, but increase their sensitivity to cisplatin-induced apoptotic death.

Mitochondrial dysfunction is known to contribute to cancer initiation, progression, and chemo-and radio-resistance. However, the precise role of mitochondria in cancer is controversial. Hence, here we tried to further clarify the role of mitochondria in cancer by transferring healthy mitochondria to cancer cells, and also to cells with depletedmitochondrialDNA (ρ0). Healthy mitochondria were isolated from WI-38 cellsand were transferred to HeLa, SAS, HeLa ρ0, and SAS ρ0 cells. Then, cell proliferation was verified. In addition, the cells were treated by different concentrations of cisplatin and assessed for apoptosis induction and quantifying the mRNA expressionof apoptosis-related genes. Results revealed that incubation of the HeLa,SAS and HeLaρ0 cells with 5µg/ml of the isolated mitochondria for 24h significantly (p < 0.001) increased cell proliferation compared to non-treated controls. Interestingly, the mitochondria transfer rescued the ρ0 cells and made them capable ofgrowing under conventional culture medium. However, the number of apoptotic cells wassignificantlyhigher in theHeLa ρ0 cells that received the mitochondria (HeLa-Fibro-Mit) compared to the HeLa ρ0. Furthermore, the expression level of BCL-2 anti-apoptotic gene was down-regulated in bothHeLa-Fibro-Mit andSAS-Fibro-Mit cell lines while the expression levels of the BAX,caspase8,caspase9, and AIFpro-apoptotic genes were upregulated. Our findings indicated that although the response of cancer cells to the mitochondria transfer is cancer-type dependent, but the introductionof normal exogenous mitochondria to some cancer cells might be considered as a potential novel therapeutic strategy.

The combination of strength and cryogenic impact toughness in low carbon 5Mn–5Ni steel

A low carbon medium manganese steel with additions of nickel and niobium was designed to further enhance cryogenic impact toughness of medium manganese steels and suppress Lüders deformation. It is interestingly found that the steel containing little retained austenite (around 1.5 vol.%) possesses high strength and excellent cryogenic impact toughness after heat treated at 650 °C for 2 h, whereas conventional medium manganese steels containing little or more unstable retained austenite always show poor cryogenic impact toughness after heat treated at a relatively high temperature. The excellent cryogenic impact toughness is mainly attributed to the formation of fine grains and little retained austenite with high mechanical stability. All steels exhibit continuous yielding behavior, and the yielding behavior is independent of heat treatment temperature. Moreover, the Lüders deformation is suppressed by restraining the formation of Cottrell atmosphere through low carbon design, further reduction in free carbon content in matrix via both the partition of carbon from matrix to austenite and the formation of NbC, some mobile dislocations and the formation of α+α′ mixture.

Ionic liquids vs conventional solvents: A comparative study in the selective catalytic oxidations promoted by oxovanadium(IV) complexes

Two oxovanadium(IV) complexes containing 4-acyl-5-pyrazolonate-κ2-O,O' bidentate ligands with different chain lengths in the acyl moiety, namely HQC6 (complex I) and HQC17 (complex II), have been synthetized and full characterized, to study their catalytic activity toward the mild and selective oxidation of olefins or model organosulphur substrates, promoted by H2O2 or tert-butyl hydroperoxide (TBHP). The influence due to the type of solvent, likewise ionic liquids (ILs) or conventional medium, was analysed. H2O2 has proven less efficient, in comparison to TBHP. Quantum-chemical calculations have shown that, the key catalytic species involved and, consequently, the actual mechanism might be slightly different according to the oxidant used, i.e. the peroxo VO(QMe)(O2) adduct in the case of H2O2 and the tert-butylperoxo VO(QMe)2(OOt-Bu) adduct in the case of TBHP. Preliminary calculations suggested that, in ionic liquids, the VO(QMe)2 complex might reveal relatively unstable, hence qualitatively explaining the moderate efficiency observed in these media.

Successful scale up expansion of Wharton's jelly mesenchymal stromal cells in different commercial xeno-free and serum-free media

Background & Aim Cell-based therapies are becoming widely accepted in regenerative medicine and their demand and potential is increasing. Human multipotent Mesenchymal Stromal Cells (MSC) derived from the Wharton's jelly (WJ) of the umbilical cords can be obtained from samples typically discarded from public cord blood banking programmes. However, current cell culture bioprocesses for the derivation, scale up expansion, and GMP-compliant product manufacturing for early phase clinical trials use serum and xenogeneic proteins in medium formulation. Therefore, the use of xeno-free and serum-free media (XSFM) formulations is required to ensure more homogenous cell populations. The aim of this study was to compare four commercially available XSFM as an alternative to human serum-containing media in WJ-MSC culture. Methods, Results & Conclusion Cryopreserved MSC derived from WJ in serum-conditions were cultured in four commercial XSFM, namely PRIME-XV MSC Expansion XSFM, MSC Nutristem® XF, MesenCult™-ACF Plus Medium, and MSC-Brew GMP Medium and were further compared with conventional expansion medium Dulbecco's modified Eagle's medium (DMEM) containing 2 mM glutamine and supplemented with 10% of human serum B (hSerB). Cells were successfully expanded in culture in all XSFM conditions. Cumulative population doublings and harvest cell densities were higher in early passages for 3 out of 4 XSFM than serum-containing expansion medium. WJ-MSC proliferation was higher than 1 × 105 cells/cm2 in early culture (passage 2) while it decreased under 1 × 105 cells/cm2 in late culture (passage 8) in XSFM conditions. Spindle shaped morphology was kept upon passages in XSFM conditions. In all cases, cells were highly positive for CD73, CD90 and CD105 and negative for CD31, CD45 and HLA-DR expression in passage 1 and 6 and multilineage capacity to osteogenic, adipogenic and chondrogenic lineages was confirmed after in vitro induction. These data demonstrate that WJ-MSC can be successfully cultured and expanded in commercially available xeno-free and serum-free media and retain all necessary characteristics attributed to MSC for potential therapeutic use.

Placenta-derived mesenchymal stem cells enhance human liver organoid maturation: translational implications for liver regeneration

Background & Aim The global prevalence of end-stage liver disease (ESLD) has reached very high levels, and, to date, the best treatment option is orthotopic liver transplantation (1). However, due to the complexity of this procedure and the limited numbers of healthy organ donors, new therapeutic options are being sought. Liver progenitor and multipotent stem cells offer potential cell sources that could be used clinically (2). Different studies have reported therapeutic effects of transplanted mesenchymal stem cells (MSCs) on ESLD (3). Indeed, an improvement of liver function has been achieved in patients with liver cirrhosis after autologous MSCs injection (4). Based on our recent work where we showed that a three-dimensional (3D) culture method is effective to improve MSCs paracrine activity (5), in this work, we used a human liver organoid system to study if the paracrine signal secreted by human amnion-derived MSCs (hAMSCs) affect liver stem/progenitor cells to differentiate (Fig. 1). Epithelial organoids recapitulate multiple aspects of real organs, making them promising models of organ function to model and treat human disease (6). Methods, Results & Conclusion Our data show that the expanded human liver organoids had distinct traits in the ALB production, Krt19 and HNF1β expression when compared to the reference cell line HepG2 cells. On liver organoids, we tested conventional differentiation medium (DM) conditioned by both 2D (2D CM) and 3D (3D CM) hAMSCs cultures observing that ALB, NTCP (markers of mature hepatocytes), and LGR5 (stem cell marker) expression were significantly upregulated by 3D CM compared with both 2D CM and conventional DM. Furthermore, CYP3A4 expression (marker of mature hepatocytes) was upregulated in both 2D and 3D CM. These data were also confirmed by the over-production/activity of both ALB and CYP3A4 observed in organoids grown with 3D CM. Liver epithelial dysfunctional repair play a role in the development of liver disease, and, effective repair likely requires the normal functioning of liver stem/progenitor cells. Herein, we showed that hAMSCs, principally grown in 3D cultures, has the potential to increase hepatic stem/progenitor cell differentiation, demonstrating that soluble factors secreted by hAMSCs are likely responsible for the response. This work revealing a potential approach to improve cell transplantation with the ultimate goal being efficient liver regeneration.

Human Cerebrospinal Fluid Induces Neuronal Excitability Changes in Resected Human Neocortical and Hippocampal Brain Slices.

Human cerebrospinal fluid (hCSF) has proven advantageous over conventional medium for culturing both rodent and human brain tissue. In addition, increased activity and synchrony, closer to the dynamic states exclusively recorded in vivo, were reported in rodent slices and cell cultures switching from artificial cerebrospinal fluid (aCSF) to hCSF. This indicates that hCSF possesses properties that are not matched by the aCSF, which is generally used for most electrophysiological recordings. To evaluate the possible significance of using hCSF as an electrophysiological recording medium, also for human brain tissue, we compared the network and single-cell firing properties of human brain slice cultures during perfusion with hCSF and aCSF. For measuring the overall activity from a majority of neurons within neocortical and hippocampal human slices, we used a microelectrode array (MEA) recording technique with 252 electrodes covering an area of 3.2 × 3.2 mm2. A second CMOS-based MEA with 4225 sensors on a 2 × 2 mm2 area was used for detailed mapping of action potential waveforms and cell identification. We found that hCSF increased the number of active electrodes and neurons and the firing rate of the neurons in the slices and induced an increase in the numbers of single channel and population bursts. Interestingly, not only an increase in the overall activity in the slices was observed, but a reconfiguration of the network could also be detected with specific activation and inactivation of subpopulations of neuronal ensembles. In conclusion, hCSF is an important component to consider for future human brain slice studies, especially for experiments designed to mimic parts of physiology and disease observed in vivo.

Preserving Airway Smooth Muscle Contraction in Precision-Cut Lung Slices

Precision-cut lung slices (PCLS) are ideal for measuring small airway contraction. However, these measurements are currently limited to acute exposure scenarios that typically last a few minutes to a few hours. Using an insulin-supplemented culture medium, we prolong the small airway contractility in mouse PCLS for up to two weeks. Compared to conventional culture medium, insulin-supplemented culture medium provides no additional benefit in preserving cellular viability or airway structure. However, it protects the airway smooth muscle (ASM) against a loss of smooth muscle myosin heavy chain (SMMHC) expression. We elucidate the significance of this new culture medium for chronic disease modeling of IL-13-induced airway hyper-responsiveness.

Palm oil mill effluent as a potential medium for microalgae Chlorella sp. cultivation for lipid production

Freshwater microalgae Chlorella sp. was isolated from the water body of Taman Gelora, Kuantan, Pahang. The isolated monoculture was cultured in different concentration of POME (10%, 20%, 30%, 40% and 50%, v/v) as substrate. The maximal growth and biomass productivity were found in 30% (v/v) of POME. The maximal dry biomass 1.56 g/L was obtained and extracted 39.1% lipid, approximating similar productivity as in conventional BG-11 medium 1.65 g/L of biomass and 41.5% lipid. Gas chromatography-mass spectrometry analysis shows the composition of fatty acids such as linolelaidic acid, oleic acid, palmitic acid, stearic acid, elaidic acid, and α-linoleic acid dominant. The highest fatty acid is linolelsidic acid (41.69%). POME has the potential to be used as a substrate for microalgae Chlorella sp. cultivation.

Sunflower Protein Enzymatic Hydrolysates as a Medium for Vitamin B2 and B12 Biosynthesis

Lowering the costs of industrial fermentation processes is a current challenge of modern biotechnology. In this sense, has been studied the possibility to replace the conventional medium for vitamin B2 abd B12 biosynthesis with new effective medium from renuable plant-based sources. Riboflavin (vitamin B2) and cobalamin (vitamin B12) are essencial compounds in humans diets and their demand is increasing globally. Current manufacturing process of these vitamins is unsustainable because of using costly unreneable resources and generating hazardous wastes. Meat peptone is conventionally used as a nitrogen source during the fermentation process, however it is relatively expencive. Sunflower protein is a secondary product, which can be obtained from sunflower meal after oil extraction, and rich on its nutritional value. The objective of this study was to select and evaluate fermentative potential of sunflower protein and its enzymatic hydrolysates as a nitrogen source for riboflavin and cobalamin biosynthesis by evaluating the kinetic parameters of strains growth, specific substrate consumprion and formation of metabolites. The fermentation of strains was carried out using conventional mineral medium with meat peptone in 250 mL Erlenmeyer flasks, and the results obtained were compared with fermentation where sunflower protein and it’s enzymatic hydrolysates were used as a nitrogen source instead of meat peptone. It was observed that when compared with meat peptone, some strains show positive effect in growth when nitrogen source was substituted with sunflower protein enzymatic hydrolysates. This allows affirming that sunflower protein and its enzymatic hydrolysates are promising in the use as a nitrogen sourse for riboflavin and cobalamin biosynthesis. The characteristics and parameters of studied fermentation processes reveals their potential for residue products applications for obtaining products with high added value.

Utilization of the acid hydrolysate of defatted Chlorella biomass as a sole fermentation substrate for the production of biosurfactant from Bacillus subtilis C9

Although algal biofuels have been promoted as a carbon-neutral alternative to fossil fuels, several economic and technological barriers challenge the much-anticipated success of algae as a commercial bio-feedstock. In particular, utilizing only a lipid portion of harvested algal biomass limits possible monetary returns from algal biomass within which non-lipid compounds generally comprise a major proportion. Previous studies thus demonstrated the production of ethanol from defatted algal biomass to improve overall economic outlook of the nascent algae industry; however, it is also possible to utilize residual algal biomass as a fermentation substrate for bacterial strains producing high value alternative products. Therefore, the goal of this study was to compare the yield of biosurfactant from B. subtilis C9 in conventional culture medium and the acid hydrolysate of defatted Chlorella biomass (DCB). Although increasing the concentration of acid catalyst from 0 N to 1.0 N was expected to proportionally enhance the effectiveness of dilute acid hydrolysis, the results indicated that DCB hydrolysates treated under different acid conditions at 121 °C for 15 min exhibited the highest yield of fermentable monosaccharides when treated with 0.25 N HCl. Subsequently, the fermentation of B. subtilis C9 was performed in both Luria-Bertani (LB) and DCB hydrolysate medium after adjusting salinity and monosaccharide concentration to identical levels; a significantly greater yield of biosurfactant was observed in the culture grown with DCB hydrolysate medium, while the presence of magnesium and manganese co-factors in DCB hydrolysate contributed to a greater biosurfactant yield. Collectively, these results suggest that the valorization of defatted algal biomass could be also accomplished by incorporating biosurfactant fermentation into algal biorefinery processes and call for further process optimization efforts that consider factors associated with the operation of industrial algal biorefineries.

Demonstration of Sputtering Atomic Layer Augmented Deposition: Aluminum Oxide-Copper Dielectric-Metal Nanocomposite Thin Films.

Designing a thin-film structure often begins with choosing a film deposition method that employs a specific process by which chemical species are formed and transported; in other words, a film deposition system in which two deposition methods are hybridized should lead to new ways of designing thin-film structures. This premise inspires us to combine atomic layer deposition (ALD) and magnetron sputtering (SPU) within a single chamber-supttering atomic layer augmented deposition (SALAD). SALAD takes full advantage of both ALD's precise and accurate precursor delivery and SPU's versatility in choosing chemical elements. A SALAD system is designed based on knowledge obtained from computational fluid dynamics with the goal of conceiving a film deposition system that satisfies deposition conditions distinctive for both ALD and SPU. As a demonstration, the SALAD system is utilized to deposit a unique nanocomposite made of aluminum oxide (Alox) thin films by ALD and copper (Cu) thin films by SPU-AlOx-Cu nanocomposite thin films. Spectroscopic reflectivity collected on AlOx-Cu nanocomposite thin films shows unique dispersion features to which conventional effective medium theories used for describing optical properties of composites made of a dielectric host that contains metallic inclusions do not seem to simply apply.