Glucose-containing Medium Research Articles

Application of glucose to prepare Aspergillus oryzae koji as an adjunct to prevent rancidity in cheese products.

Koji made using Aspergillus oryzae shows potential for application as a cheese adjunct; however, flavor defects resulting from volatile free fatty acid (FFA) accumulation should be avoided. Hence, a modified glucose-containing whey solid medium was used to culture A. oryzae AHU 7139, and the triacylglycerol (TG) lipase activity and lipase gene (tglA and mdlB) expression were compared with those of a culture using a conventional whey solid medium. The results showed that TG lipase activity and the expression of both lipase genes were reduced in the modified medium. Moreover, the expression level of farA, a positive transcription factor of the lipase genes, was also reduced. The cheese adjunct prepared by culturing the AHU 7139 strain in the modified medium lowered the FFA content in the cheese products, resulting in comparable FFA levels with those in the adjunct-free cheese. Thus, adding glucose is recommended to prepare the koji adjunct for cheesemaking.

Culture of Bovine Aortic Endothelial Cells in Galactose Media Enhances Mitochondrial Plasticity and Changes Redox Sensing, Altering Nrf2 and FOXO3 Levels.

Understanding the complex biological processes of cells in culture, particularly those related to metabolism, can be biased by culture conditions, since the choice of energy substrate impacts all of the main metabolic pathways. When glucose is replaced by galactose, cells decrease their glycolytic flux, working as an in vitro model of limited nutrient availability. However, the effect of these changes on related physiological processes such as redox control is not well documented, particularly in endothelial cells, where mitochondrial oxidation is considered to be low. We evaluated the differences in mitochondrial dynamics and function in endothelial cells exposed to galactose or glucose culture medium. We observed that cells maintained in galactose-containing medium show a higher mitochondrial oxidative capacity, a more fused mitochondrial network, and higher intercellular coupling. These factors are documented to impact the cellular response to oxidative stress. Therefore, we analyzed the levels of two main redox regulators and found that bovine aortic endothelial cells (BAEC) in galactose media had higher levels of FOXO3 and lower levels of Nrf2 than those in glucose-containing media. Thus, cultures of endothelial cells in a galactose-containing medium may provide a more suitable target for the study of in vitro mitochondrial-related processes than those in glucose-containing media; the medium deeply influences redox signaling in these cells.

Astrocyte Mitochondria Are a Sensitive Target of PCB52 and its Human-Relevant Metabolites.

Polychlorinated biphenyls (PCBs) are industrial chemicals that are ubiquitously found in the environment. Exposure to these compounds has been associated with neurotoxic outcomes; however, the underlying mechanisms for such outcomes remain to be fully understood. Recent studies have shown that astrocytes, the most abundant glial cell type in the brain, are susceptible to PCB exposure as well as exposure to human-relevant metabolites of PCBs. Astrocytes are critical for maintaining healthy brain function due to their unique functional attributes and positioning within the neuronal networks in the brain. In this study, we assessed the toxicity of PCB52, one of the most abundantly found PCB congeners in outdoor and indoor air, and two of its human-relevant metabolites, on astrocyte mitochondria. We exposed C6 cells, an astrocyte cell line, to PCB52 or its human-relevant metabolites and found that all the compounds showed increased toxicity in galactose-containing media compared to that in the glucose-containing media, indicating the involvement of mitochondria in observed toxicity. Additionally, we also found increased oxidative stress upon exposure to PCB52 metabolites. All three compounds caused a loss of mitochondrial membrane potential, distinct changes in the mitochondrial structure, and impaired mitochondrial function. The hydroxylated metabolite 4-OH-PCB52 likely functions as an uncoupler of mitochondria. This is the first study to report the adverse effects of exposure to PCB52 and its human-relevant metabolites on the mitochondrial structure and function in astrocytes.

Effect of quercetin on oxidative stress in 3T3-L1 mature and hypertrophic cells

Abstract Purpose: The process of excessive or abnormal accumulation of fat in the body is called obesity, and its prevalence is increasing globally. The imbalance between antioxidants and free radicals, or oxidative stress, can be caused by or result from obesity. Flavonoids with antioxidant potential may help lower the increased oxidative stress associated with obesity. This study aimed to determine how quercetin affected oxidative stress in hypertrophied and mature 3T3-L1 adipocytes. Materials and methods: After differentiating, 3T3-L1 adipocytes were treated with insulin and a glucose-containing medium to become mature (10 days) and hypertrophic (18 days). The cells were subsequently incubated with 80 µM quercetin for 24 and 48 hours. ELISA was used to determine the levels of total antioxidant capacity/total oxidant capacity (TAS/TOS). Using Oil Red O staining, an accumulation of triglycerides in cells was examined. Results: It was shown that the quercetin molecule had a prooxidative effect on hypertrophic adipocytes but did not impact on oxidative stress in mature adipocytes. Conclusion: It is believed that the number and kind of fat cell will determine the appropriate dose and duration of administration for quercetin's antioxidant mechanism of action, which produces numerous beneficial effects.

Caldanaerobacter subterraneus subsp. keratinolyticus subsp. nov., a Novel Feather-Degrading Anaerobic Thermophile.

Caldanaerobacter subterraneus subsp. keratinolyticus subsp. nov. strain KAk was isolated from a geothermal hot spring located in Kazakhstan. Growth occurred at temperatures ranging from 50 to 80 °C, with approximately 70 °C as optimum. It also thrived in pH conditions ranging from 4.0 to 9.0, with the best growth occurring at 6.8. Under optimal conditions in a glucose-containing medium, the cells were predominantly observed singly, in pairs, or less frequently in chains, and did not form endospores. However, under conditions involving growth with merino wool or feathers, or under suboptimal conditions, the cells of strain KAk exhibited a notably elongated and thinner morphology, with lengths ranging from 5 to 8 µm, and spores were observed. The KAk strain exhibited efficient degradation of feather keratin and merino wool at temperatures ranging from 65 to 70 °C. Analysis of the 16S rRNA gene sequence placed KAk within the genus Caldanaerobacter, family Thermoanaerobacteraceae, with the highest similarity to C. subterraneus subsp. tengcongensis MB4T (98.84% sequence identity). Furthermore, our analysis of the draft genome sequence indicated a genome size of 2.4 Mbp, accompanied by a G+C value of 37.6 mol%. This study elucidated the physiological and genomic characteristics of strain KAk, highlighting its keratinolytic capabilities and distinctiveness compared to other members of the genus Caldanaerobacter.

Effect of dehydration method on the micro- and nanomorphological properties of bacterial cellulose produced by Medusomyces gisevii on different substrates

Many important properties of bacterial cellulose (BC), such as moisture absorption capacity, elasticity and tensile strength, largely depend on its structure. This paper presents a study on the effect of the drying method on BC films produced by Medusomyces gisevii using two different procedures: room temperature drying (RT, (24 ± 2 °C, humidity 65 ± 1%, dried until a constant weight was reached) and freeze-drying (FD, treated at − 75 °C for 48 h). BC was synthesized using one of two different carbon sources—either glucose or sucrose. Structural differences in the obtained BC films were evaluated using atomic force microscopy (AFM), scanning electron microscopy (SEM), and X-ray diffraction. Macroscopically, the RT samples appeared semi-transparent and smooth, whereas the FD group exhibited an opaque white color and sponge-like structure. SEM examination showed denser packing of fibrils in FD samples while RT-samples displayed smaller average fiber diameter, lower surface roughness and less porosity. AFM confirmed the SEM observations and showed that the FD material exhibited a more branched structure and a higher surface roughness. The samples cultivated in a glucose-containing nutrient medium, generally displayed a straight and ordered shape of fibrils compared to the sucrose-derived BC, characterized by a rougher and wavier structure. The BC films dried under different conditions showed distinctly different crystallinity degrees, whereas the carbon source in the culture medium was found to have a relatively small effect on the BC crystallinity.Graphical

Bioconversion of Shrimp Waste into Functional Lipid by a New Oleaginous Sakaguchia sp.

Chitin, the second most abundant biomolecule after cellulose in nature, is a significant aquaculture by-product, and is estimated at 6-8 million tons annually. Chitin is composed of monomeric N-acetylglucosamine (NAG) which can be seen as an alternative feedstock for biotechnology. Microbial functional lipids have gained attention due to their bioactivity and sustainable production. In this study, a new oleaginous yeast strain named Sakaguchia sp. HKC2 was found to be able to use NAG as the carbon source for growth and accumulate functional lipids such as PUFAs and carotenoids. When cultured on the NAG-containing medium, strain HKC2 exhibited slower growth and slower intracellular lipid accumulation compared to those on a glucose-containing medium. However, the lipids obtained from HKC2 grown on NAG medium were richer in PUFAs. Notably, torularhodin-a powerful bioactive carotenoid-was found in all HKC2 cultures on NAG, while torulene was abundant in glucose medium. These findings highlight a novel avenue for utilizing aquatic by-products and unlocking their potential.

Grazing resistance developed in Escherichia coli K-12 during coexistence with a bacterivorous protist.

A development of grazing resistance in Escherichia coli K-12 was examined in the presence of a bacterivorous protist, Spumella sp. TGKK2. Two transformants were generated from E. coli K12 for grazing experiments. One was E. coli K-12-TGF, which possesses tetracycline resistance and green fluorescence. The other was E. coli K-12-KRF with kanamycin resistance and red fluorescence. These strains can be selectively colonized on antibiotic-containing agar media and further confirmed by their fluorescent colors. First, we added protist-untouched E. coli K-12-KRF to protist-touched residual E. coli K-12-TGF that had been attacked by Spumella sp. TGKK2 in a batch test. Then the survivability of the respective strains was investigated. Consequently, E. coli K-12-KRF was predated preferentially. On the other hand, E. coli K-12-TGF in the same tube was less predated, indicating some grazing resistance. Similar phenomena were observed when the conditions of these two strains of bacteria were reversed. Also, a continuous culture device supplied with a glucose-containing medium as a substrate was operated. The device connected two complete mixed reactors in series. E. coli K-12-TGF was cultivated in the first reactor, and then grown E. coli K-12-TGF was predated by Spumella sp. TGKK2 in the second reactor. The effluent in the second reactor containing residual E. coli K-12-TGF and Spumella sp. TGKK2 was supplemented with batch-cultured E. coli K-12-KRF. Consequently, it was confirmed that bach-cultured E. coli K-12-KRF never exposed to protist was predated preferentially. These findings reveal that E. coli K12 acquires some predation resistance through coexistence with the bacterivorous protist.

An in vitro Model to Study Placental Functions in Gestational Diabetes Mellitus

Background: Gestational diabetes mellitus (GDM) is a serious pregnancy complication that affects around 14 % of pregnancies worldwide. It not only causes short-term complications in the mother and child but also significantly increases the risks of several chronic diseases, including cardiovascular diseases and diabetes. An excess production of the soluble vascular endothelial growth factor (VEGF) receptor 1 in the placenta, which is a natural inhibitor of VEGF, has been linked to various pregnancy complications. However, placentas affected by GDM often exhibit increased vascularization, suggesting that the role of VEGF in GDM differs from its impact on typical pregnancy complications. Unfortunately, limited studies on placental functions in GDM have been conducted due to a lack of reliable culture models. Methods: In this study, we developed an in vitro model of GDM using the human choriocarcinoma trophoblast cell line BeWo and examined the effects of VEGF on glucose uptake. The BeWo cells were treated with a high glucose-containing (25 mM) syncytialization differentiation medium to produce insulin-resistant cells. Insulin resistance was assessed by a glucose uptake assay using the fluorescent glucose analog 2-NBDG and flow cytometry analysis. Results: The cells treated with high glucose exhibited a significant decrease in glucose uptake, suggesting the successful development of glucose resistance in these cells. However, VEGF treatment did not show a significant impact on glucose uptake in high glucose-treated cells. Conclusions: It is important to note that glucose homeostasis is a complex process involving multiple cell types, and further studies are necessary to fully understand the functions of VEGF on GDM placentas. Nevertheless, the in vitro model we have developed and validated will be a valuable tool for studying placental pathophysiology in GDM and evaluating the effectiveness of potential therapeutic agents. HIGHLIGHTS BeWo cells are the choriocarcinoma cell line which were cultured in the in vitro condition BeWo cells were differentiated using forskolin and 8-Br Camp Insulin resistant model of BeWo cells were prepared by high glucose (25 mM) treatment The effect of VEGF on glucose homeostasis in GDM model of BeWo cells was observed GRAPHICAL ABSTRACT

Production of Lactic Acid by lactobacilli using Legume Flours

Due to their high nutritional value, richness in carbohydrates and good source of protein, legumes are considered essential for human diet. Legumes flours were used as fermentable material for lactic acid production. Hence, previously isolated lactobacilli strains were used in this study. The strains showed strong microbial growth and their survival in glucose-containing MRS medium and were described using a modified Gompertz equation. Lactobacilli exhibited the shortest latency phase in MRS-glucose medium. While the highest lactic acid produced was 15.40g/l by Lpb. pentosus U1 isolate after 48hours. Furthermore, these strains were evaluated in flours samples for acidification capacity. The results showed a decrease in pH values after 12hours of fermentation, indicating the fastest acidification with maximum lactic acid generation of 31.66, 25.33, 31.00 and 31.66g/l for black gram flour by Lpb. plantarum M1, Lpb. pentosus U1, Lpb. plantarum VIP1 and Lev. brevis TIP1 isolates. Black gram flour was considered the best among the used substrates for enhancing lactic acid production. Overall, this study can be termed as a preliminary step for fermentation using the desired organism in preparing legume-based food at the industrial level and benefiting consumers.

Identification and characterization of sugar-regulated promoters in Chaetomium thermophilum

The thermophilic fungus Chaetomium thermophilum has been used extensively for biochemical and high-resolution structural studies of protein complexes. However, subsequent functional analyses of these assemblies have been hindered owing to the lack of genetic tools compatible with this thermophile, which are typically suited to other mesophilic eukaryotic model organisms, in particular the yeast Saccharomyces cerevisiae. Hence, we aimed to find genes from C. thermophilum that are expressed under the control of different sugars and examine their associated 5’ untranslated regions as promoters responsible for sugar-regulated gene expression. To identify sugar-regulated promoters in C. thermophilum, we performed comparative xylose- versus glucose-dependent gene expression studies, which uncovered a number of enzymes with induced expression in the presence of xylose but repressed expression in glucose-supplemented media. Subsequently, we cloned the promoters of the two most stringently regulated genes, the xylosidase-like gene (XYL) and xylitol dehydrogenase (XDH), obtained from this genome-wide analysis in front of a thermostable yellow fluorescent protein (YFP) reporter. With this, we demonstrated xylose-dependent YFP expression by both Western blotting and live-cell imaging fluorescence microscopy. Prompted by these results, we expressed the C. thermophilum orthologue of a well-characterized dominant-negative ribosome assembly factor mutant, under the control of the XDH promoter, which allowed us to induce a nuclear export defect on the pre-60S subunit when C. thermophilum cells were grown in xylose- but not glucose-containing medium. Altogether, our study identified xylose-regulatable promoters in C. thermophilum, which might facilitate functional studies of genes of interest in this thermophilic eukaryotic model organism.

Unraveling the Molecular Basis of Mycosporine Biosynthesis in Fungi.

The Phaffia rhodozyma UCD 67-385 genome harbors a 7873 bp cluster containing DDGS, OMT, and ATPG, encoding 2-desmethy-4-deoxygadusol synthase, O-methyl transferase, and ATP-grasp ligase, respectively, of the mycosporine glutaminol (MG) biosynthesis pathway. Homozygous deletion mutants of the entire cluster, single-gene mutants, and the Δddgs-/-;Δomt-/- and Δomt-/-;Δatpg-/- double-gene mutants did not produce mycosporines. However, Δatpg-/- accumulated the intermediate 4-deoxygadusol. Heterologous expression of the DDGS and OMT or DDGS, OMT, and ATPG cDNAs in Saccharomyces cerevisiae led to 4-deoxygadusol or MG production, respectively. Genetic integration of the complete cluster into the genome of the non-mycosporine-producing CBS 6938 wild-type strain resulted in a transgenic strain (CBS 6938_MYC) that produced MG and mycosporine glutaminol glucoside. These results indicate the function of DDGS, OMT, and ATPG in the mycosporine biosynthesis pathway. The transcription factor gene mutants Δmig1-/-, Δcyc8-/-, and Δopi1-/- showed upregulation, Δrox1-/- and Δskn7-/- showed downregulation, and Δtup6-/- and Δyap6-/- showed no effect on mycosporinogenesis in glucose-containing medium. Finally, comparative analysis of the cluster sequences in several P. rhodozyma strains and the four newly described species of the genus showed the phylogenetic relationship of the P. rhodozyma strains and their differentiation from the other species of the genus Phaffia.

Heat resistance acquirement of the spoilage yeast Saccharomyces diastaticus during heat exposure

The main fungal cause of spoilage of carbonated fermented beverages in the brewing industry is the amylolytic budding yeast Saccharomyces cerevisiae subsp. diastaticus (Saccharomyces diastaticus). Heat treatment is used to avoid microbial spoilage of the fermented beverages. Therefore, the spoilage capacity of S. diastaticus may be linked to its relative high heat resistance. Here, we assessed whether S. diastaticus can acquire heat resistance when exposed to heat stress. To this end, ascospores of S. diastaticus strain MB523 were treated at 60°C for 10 min followed by growing the surviving spores on a glucose-containing medium. The resulting vegetative cells were then allowed to sporulate again in sporulation medium. This cycle of heat treatment, vegetative growth, and sporulation was performed eight times in three independent lineages. After these eight cycles, the sporulation rate was similar to the start (∼75%) but the resulting ascospores were more heat resistant. The time needed to kill 90% of the population at 60°C (i.e. the D60-value) increased from 6.5 to 9.0 min (p = 0.005). The vegetative cells also showed a trend to increased heat resistance with an increase in the D52-value from 9.2 to 16.2 min (p = 0.1). In contrast, heat resistance of the vegetative cells that had not been exposed to heat during the eight cycles had been reduced with a D52-value of 4.2 min (p = 0.003). Together, these data show that S. diastaticus MB523 can easily acquire heat resistance by inbreeding while subjected to heat stress. Conversely, heat resistance can be easily lost in the absence of this stress condition, indicative of a trade-off for heat resistance.

1,25-dihydroxyvitamin D3 ameliorates high glucose-mediated proliferation, migration, and MCP-1 secretion of vascular smooth muscle cells by inhibiting MAPK phosphorylation

ObjectivesTo explore the impacts of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on the proliferation, migration, and monocyte chemoattractant protein-1 (MCP-1) secretion of vascular smooth muscle cells (VSMCs) in a high glucose environment and its possible mechanism.MethodsWe extracted VSMCs from the thoracic aorta of a male Sprague–Dawley rats before culturing them in a 25-mM glucose-containing medium in the presence or absence of 1,25(OH)2D3 (10−9 –10−7 M). Cell proliferation was determined by bromodeoxyuridine incorporation assays. Subsequently, cell migratory capacity was examined by performing a transwell assay. An enzyme-linked immunosorbent assay was conducted to assess MCP-1 levels. Protein levels of matrix metalloproteinase-9 (MMP-9), mitogen-activated protein kinases (MAPKs), cyclin D1, and phosphorylated MAPKs were determined by immunoblotting.Results1,25(OH)2D3 significantly suppressed the proliferation, migration, and MCP-1 secretion of VSMCs mediated by high glucose in a dose-dependent manner, diminished the enhanced protein expression of MMP-9 and cyclin D1, and attenuated MAPK phosphorylation. The p38 inhibitor SB203580 and ERK1/2 inhibitor PD98059 suppressed high glucose-mediated upregulation of MMP-9 and cyclin D1 protein expression and MCP-1 secretion, respectively.Conclusions1,25(OH)2D3 ameliorates high glucose-mediated proliferation, migration, and MCP-1 secretion of VSMCs by inhibiting MAPK phosphorylation, implying a potential therapeutic approach using 1,25(OH)2D3 for diabetic macrovascular complications.

Amino Acids Supplied through the Autophagy/Endocytosis Pathway Promote Starch Synthesis in Physcomitrella Protonemal Cells.

The physiological implications of autophagy in plant cells have not been fully elucidated. Therefore, we investigated the consequences of autophagy in the moss Physcomitrella by measuring biochemical parameters (fresh and dry weights; starch, amino acid, carbohydrate, and NH3 content) in wild-type (WT) and autophagy-deficient atg5 Physcomitrella cells. We found higher starch levels and a higher net starch synthesis rate in WT cells than in atg5 cells cultured in a glucose-containing culture medium, whereas net starch degradation was similar in the two strains cultured in a glucose-deficient culture medium. Additionally, the treatment of cells with the autophagy inhibitor 3-methyladenine suppressed starch synthesis. Loading bovine serum albumin into atg5 cells through endocytosis, i.e., supplying proteins to vacuoles in the same way as through autophagy, accelerated starch synthesis, whereas loading glutamine through the plasma membrane had no such effect, suggesting that Physcomitrella cells distinguish between different amino acid supply pathways. After net starch synthesis, NH3 levels increased in WT cells, although the change in total amino acid content did not differ between WT and atg5 cells, indicating that autophagy-produced amino acids are oxidized rapidly. We conclude that autophagy promotes starch synthesis in Physcomitrella by supplying the energy obtained by oxidizing autophagy-produced amino acids.

Glucose deprivation reduces proliferation and motility, and enhances the anti-proliferative effects of paclitaxel and doxorubicin in breast cell lines in vitro.

Breast cancer chemotherapy with high dose alkylating agents is severely limited by their collateral toxicity to crucial normal tissues such as immune and gut cells. Taking advantage of the selective dependence of cancer cells on high glucose and combining glucose deprivation with these agents could produce therapeutic synergy. In this study we examined the effect of glucose as well as its deprivation, and antagonism using the non-metabolized analogue 2-deoxy glucose, on the proliferation of several breast cancer cell lines MCF7, MDA-MB-231, YS1.2 and pII and one normal breast cell line, using the MTT assay. Motility was quantitatively assessed using the wound healing assay. Lactate, as the end product of anaerobic glucose metabolism, secreted into culture medium was measured by a biochemical assay. The effect of paclitaxel and doxorubicin on cell proliferation was tested in the absence and presence of low concentrations of glucose using MTT assay. In all cell lines, glucose supplementation enhanced while glucose deprivation reduced both their proliferation and motility. Lactate added to the medium could substitute for glucose. The inhibitory effects of paclitaxel and doxorubicin were significantly enhanced when glucose concentration was decreased in the culture medium, requiring 1000-fold lesser concentration to achieve a similar degree of inhibition to that seen in glucose-containing medium. Our data show that a synergy was obtained by combining paclitaxel and doxorubicin with glucose reduction to inhibit cancer cell growth, which in vivo, might be achieved by applying a carbohydrate-restricted diet during the limited phase of application of chemotherapy; this could permit a dose reduction of the cytotoxic agents, resulting in greater tolerance and lesser side effects.

Effect of added sulphur dioxide levels on the aroma characteristics of wines from Panciu wine region

In the wine-making process, the yeasts form over 20 higher alcohols that improve the sensation of palatine veil for the wine. The amount of higher alcohols is lower for white and rosé wines obtained from clarified musts fermented at moderate temperatures. The formation of higher alcohols is confirmed by the fact that the addition of certain nitrogen compounds to a glucose-containing culture medium leads, after the fermentation, to an increase in the higher alcohol content. Data evaluate by comparison the levels of higher alcohols, esters and aldehydes, important to fermentation process in case of several wines from Panciu region. The wine-making was done by classical methods and by using specific protocols for low content sulphur dioxide. This study is focused to compare wines obtained from two grape varieties produced at industrial level: Feteasca regala vs. Feteasca regala Frizzante and Cabernet Sauvignon rosé vs. Cabernet Sauvignon. The compounds were analyzed by GC-FID method with head-space technique for methanol, 1-propanol, 2-methyl-propanol, 1-butanol, ethyl acetate, ethyl lactate, isoamyl acetate and acetaldehyde. The results showed that in almost every comparative study the levels of aroma compounds are maintained, when compared with the wines with low content of sulphur dioxide. Tuckey test showed that 1-propanol and ethyl acetate register statistically significant differences for Cabernet Sauvignon rosé and for Feteasca regala Frizzante.

Effects of potential inhibitors present in dilute acid-pretreated corn stover on fermentative hydrogen production by Escherichia coli

This study reports the inhibitory compounds present in dilute sulfuric acid-pretreated corn stover for fermentative hydrogen production by Escherichia coli. The E. coli W3110-derived strain deficient in lactate and succinate production pathways was used. When dilute acid-treated corn stover liquor containing 175 mM acetate, 21 mM furfural, and 2.6 mM 5-hydroxymethylfurfural was added at fourfold dilution to the xylose-containing culture medium, the microbial growth and hydrogen production were almost completely inhibited. The addition of acetate, furfural, or 5-hydroxymethylfurfural, at up to 200, 80, or 40 mM, respectively, to the xylose- or glucose-containing medium led to dose-dependent inhibition of fermentative hydrogen production. No synergistic inhibition by furfural with acetate or 5-hydroxymethylfurfural at concentrations based on the corn stover hydrolysate was observed. Altogether, these results suggest that the inhibition of the E. coli-based hydrogen production by the corn stover hydrolysate is primarily attributed to acetate under the conditions used.

Heterozygous p.Y955C mutation in DNA polymerase γ leads to alterations in bioenergetics, complex I subunit expression, and mtDNA replication

In human cells, ATP is generated using oxidative phosphorylation machinery, which is inoperable without proteins encoded by mitochondrial DNA (mtDNA). The DNA polymerase gamma (Polγ) repairs and replicates the multicopy mtDNA genome in concert with additional factors. The Polγ catalytic subunit is encoded by the POLG gene, and mutations in this gene cause mtDNA genome instability and disease. Barriers to studying the molecular effects of disease mutations include scarcity of patient samples and a lack of available mutant models; therefore, we developed a human SJCRH30 myoblast cell line model with the most common autosomal dominant POLG mutation, c.2864A>G/p.Y955C, as individuals with this mutation can present with progressive skeletal muscle weakness. Using on-target sequencing, we detected a 50% conversion frequency of the mutation, confirming heterozygous Y955C substitution. We found mutated cells grew slowly in a glucose-containing medium and had reduced mitochondrial bioenergetics compared with the parental cell line. Furthermore, growing Y955C cells in a galactose-containing medium to obligate mitochondrial function enhanced these bioenergetic deficits. Also, we show complex I NDUFB8 and ND3 protein levels were decreased in the mutant cell line, and the maintenance of mtDNA was severely impaired (i.e., lower copy number, fewer nucleoids, and an accumulation of Y955C-specific replication intermediates). Finally, we show the mutant cells have increased sensitivity to the mitochondrial toxicant 2′-3′-dideoxycytidine. We expect this POLG Y955C cell line to be a robust system to identify new mitochondrial toxicants and therapeutics to treat mitochondrial dysfunction.

Liquid-crystalline ordering in bacterial cellulose produced by Gluconacetobaсter hansenii on glucose-containing media

This research is dedicated to the studies of the microscale morphology of bacterial cellulose (BC) obtained by means of static cultivation of Gluconacetobacter hansenii GH-1/2008. We found that the microscale morphology depended on the BC production rate that was varied by using different glucose concentrations in the cultivation medium. It was revealed that at higher production rates, BC fibrils were aligned in a liquid-crystalline-like (LC-like) order. The observed helical alignment was always left-handed. The half-periods of the helix varied from 50 μm to 150 μm depending on the cultivation conditions. The mechanical and water absorption properties of the obtained BC pellicles were measured. The former correlated mainly with the density of the samples; the latter were the best for films with layered structure, where the BC had segregated into fleece sheets separated by gaps with low density of fibrils.