High Cell Concentrations Research Articles

IMPACT OF FOLIAR SPRAY WITH SACCHAROMYCES CEREVISIAE EXTRACT IN DIFFERENT FORMS AND CONCENTRATIONS ON GROWTH, PRODUCTIVITY AND QUALITY OF LETTUCE PLANTS

This investigation was adopted during the two consecutive winter seasons of 2019/2020 and 2020/2021, to examine the impact of foliar spray with bread yeast in different forms and concentrations on the development, production yield and quality of lettuce cv. Dark Green (Lactuca sativa L.). This experiment was consisted of twelve treatments which were the combinations between four yeast forms (untreated yeast cells, yeast extract, yeast cells treated with nitric acid 50% and yeast cells treated with urea at three concentrations (1, 2 and 3, w/v).The results showed that, foliar spraying with yeast cells treated with urea at the highest concentration 3% recorded the largest values of the height of plant, leaf area per plant, total fresh and dry weights/plant, total chlorophyll, yield and its components, N, K and dry matter percentages in lettuce leaves in both seasons, with no significant differences between foliar spray with yeast cells treated with urea or with nitric acid treatments regarding number of leaves/plant, P (%) and nitrate content. In general, foliar spraying with yeast cells treated with urea at 3% increased yield and its component but at the same time increased nitrate content in leaves and decreased the quality of lettuce plants. Accordingly, it may be prudent to avoid using a high concentration of yeast cells treated with urea, in order to preserve the quality of lettuce plants, and instead of that it can be used a medium concentration to get the best quality and reasonable yield.

Antibacterial activity in three Chaetoceros microalgae species cultures by using antibiotics

Diatoms, such as Chaetoceros, grow in a mutualistic relationship with bacteria. However, in some cases, it is necessary to grow them in bacteria-free cultures. To reduce bacterial load, antibiotics are used, and on certain occasions it is necessary to use a mixture with more than one antibiotic. This work aimed to obtain a quick and effective protocol to reduce the bacterial load and evaluate the response of three Chaetoceros species with aquacultural importance. Single and mix antibiotics were used. Microalgal and bacterial growth was measured. The growth parameters for diatoms showed that the significantly highest cell concentration was for C. muelleri (3.15 x106 cells mL-1) and the lowest values to C. calcitrans (2.98 x106 cells mL-1). The significantly highest growth rate was for C. calcitrans (0.77 divisions per day), and the lowest values for Chaetoceros sp. (0.60 divisions per day). The growth parameters for heterotrophic bacteria showed that the significantly highest bacterial load was for Chaetoceros sp. (19.16 x106 CFU (Colony-Forming Units) mL-1) and the lowest values were for C. calcitrans (12.23 x106 CFU mL-1). The growth rate of the heterotrophic bacteria present in Chaetoceros cultures was similar among the three studied species. Streptomycin® and sulfate G41® produced a partial reduction of bacterial load. The most effective treatment for all three species was the use of an antibiotic mix composed of ampicillin® (250 μg mL-1), kanamycin® (200 μg mL-1), neomycin® (50 μg mL-1), and streptomycin® (100 μg mL-1) for three days. The mix prepared with the highest antibiotic concentration produced a reduction of bacteria (100%) for three days; however, it also induced a significant reduction of the growth of the three Chaetoceros species.

Spermine and Spermidine Detection through Restricted Intramolecular Rotations in a Tetraphenylethylene Derivative

Biogenic polyamines, especially spermine and spermidine, are associated with cell growth and development. These amines can be found at high concentrations in the tumor cells, tissues, and urine of cancer patients. In contrast, spermidine levels drop with age, and a possible connection between low endogenous spermidine concentrations and age-related deterioration has been suggested. Thus, the quantification of these amines in body fluids like urine could be used in the diagnosis of different pathological situations. Here a new fluorescent molecular probe based on a tetraphenylethylene derivative is reported. This probe is able to selectively detect these amines through the enhancement of the fluorescence emission of the resulting complex. This fluorescence enhancement may be related to restricted intramolecular rotations of TPE phenyl rings induced by the analyte. Theoretical studies were carried out to shed light on the observed selectivity. Finally, the detection of these amines in urine was performed with limits of detection of 0.70 µM and 1.17 µM for spermine and spermidine, respectively.

Production of Recombinant RNA in Escherichia coli Using Eggplant Latent Viroid as a Scaffold.

Similar to viruses, viroids can also be engineered and transformed into useful biotechnological tools. We describe here a viroid-based system to produce large amounts of recombinant RNA in Escherichia coli. A precursor of eggplant latent viroid (ELVd), with the RNA of interest inserted between positions U245 and U246, is co-expressed in E. coli along the chloroplastic isoform of the eggplant tRNA ligase, the enzyme that mediates the circularization of this viroid in the infected plants. In the bacterial cells, the chimeric ELVd-RNA-of-interest precursor self-cleaves through the embedded hammerhead ribozymes, and the monomer is recognized and circularized by the co-expressed tRNA ligase. The resulting circular RNA, likely bound to the tRNA ligase, accumulates to a high concentration in the bacterial cells.

A designed moderately thermophilic consortia with a better performance for leaching high grade fine lead-zinc sulfide ore

Unwieldy fine sulfide ores are produced during mining; without being appropriately disposed of, they can cause environmental pollution and waste resources. This study investigated the leaching performance of a moderately thermophilic consortia (Leptospirillum ferriphilum + Acidithiobacillus caldus + Sulfobacillus benefaciens) for fine lead-zinc sulfide raw ore. The results showed this microbial community created a low pH, high ORP, and high cell concentration environment for mineral leaching, improving bioleaching efficiency. Under the action of this consortia, the zinc leaching rate reached 96.44 in 8 days, and reached 100% after 12 days. EPS analysis indicated that the consortia could mediate the secretion of more polysaccharides to ensure leaching efficiency. EPS levels and amino acids were the main factors affecting bioleaching. An analysis of mineral surface characteristics showed the consortia effectively leached pyrite and sphalerite from the fine sulfide ore, and prevented the mineral surface forming the jarosite that could hinder bioleaching. This study found that bioleaching reduced the potential environmental toxicity of the minerals, providing an important reference for guiding the bioleaching of unwieldy fine sulfide raw ore.

Expanding the Reactive Sulfur Metabolome: Intracellular and Efflux Measurements of Small Oxoacids of Sulfur (SOS) and H2S in Human Primary Vascular Cell Culture.

Hydrogen sulfide (H2S) is an endogenous signaling molecule which is important for cardiovascular health, but its mechanism of action remains poorly understood. Here, we report measurements of H2S as well as its oxidized metabolites, termed small oxoacids of sulfur (SOS = HSOH and HOSOH), in four human primary vascular cell lines: smooth muscle and endothelial cells derived from both human arterial and coronary tissues. We use a methodology that targets small molecular weight sulfur species; mass spectrometric analysis allows for species quantification to report cellular concentrations based on an H2S calibration curve. The production of H2S and SOS is orders of magnitude higher in smooth muscle (nanomolar) as compared to endothelial cell lines (picomolar). In all the primary lines measured, the distributions of these three species were HOSOH >H2S > HSOH, with much higher SOS than seen previously in non-vascular cell lines. H2S and SOS were effluxed from smooth muscle cells in higher concentrations than endothelial cells. Aortic smooth muscle cells were used to examine changes under hypoxic growth conditions. Hypoxia caused notable increases in HSOH and ROS, which we attribute to enhanced sulfide quinone oxidase activity that results in reverse electron transport.

Potential of Recombinant Raw Starch‐Degrading Enzyme from Escherichia coli for Sugar Syrup and Bioethanol Productions Using Broken Rice Powder as Substrate

AbstractThe recombinant raw starch‐degrading enzyme (LsA175) which harbors the lsa175 gene from Laceyella sacchari LP175 is optimized for expression in Escherichia coli BL21 (DE3). The recombinant strain provides an alternative avenue for enzyme production in a stirrer fermenter with tolerance to shear force and avoids losing stability by protease existing in the native strain. The maximum enzyme production at 4472±112 U g−1 cell dry weight is obtained using the optimum conditions at 22 °C, 1.0 mM IPTG, and 24 h of induction temperature, Isopropyl β‐ d‐1‐thiogalactopyranoside (IPTG) concentration, and induction time, respectively. The enzyme production of 4545 ± 89.8 U g−1 cell dry weight is obtained by batch fermentation in a 2.0 L stirrer fermenter and 5329 ± 86.1 U g−1 cell dry weight by continued glucose feeding with high bacterial cell concentration. The uncooked broken rice powder (200 g L−1) is hydrolyzed by crude recombinant LsA175 (300 U mL−1) at 50 °C for 12 h without the addition of exogenous glucoamylase (GA), yielding 86 ± 1.79 g L−1 reducing sugar. The bioethanol production resulting from modified simultaneous saccharification and fermentation with Kluyveromyces marxianus DMKU‐KS07 at 42 °C gave 71.4±4.35 g L−1 ethanol with 88.24% of theoretical ethanol yield. LsA175 presents engaging performances regarding sugar syrup and bioethanol production, which can diminish the cost of production and energy consumption.

Uric Acid as a Photosensitizer in the Reaction of Deoxyribonucleosides with UV Light of Wavelength Longer than 300 nm: Identification of Products from 2'-Deoxycytidine.

DNA reacts directly with UV light with a wavelength shorter than 300 nm. Although ground surface sunlight includes little of this short-wavelength UV light due to its almost complete absorption by the atmosphere, sunlight is the primary cause of skin cancer. Photosensitization by endogenous substances must therefore be involved in skin cancer development mechanisms. Uric acid is the final metabolic product of purines in humans, and is present at relatively high concentrations in cells and fluids. When a neutral mixed solution of 2'-deoxycytidine, 2'-deoxyguanosine, thymidine, and 2'-deoxyadenosine was irradiated with UV light with a wavelength longer than 300 nm in the presence of uric acid, all the nucleosides were consumed in a uric acid dose-dependent manner. These reactions were inhibited by the addition of radical scavengers, ethanol and sodium azide. Two products from 2'-deoxycytidine were isolated and identified as N4-hydroxy-2'-deoxycytidine and N4,5-cyclic amide-2'-deoxycytidine, formed by cycloaddition of an amide group from uric acid. A 15N-labeled uric acid, uric acid-1,3-15N2, having two 14N and two 15N atoms per molecule, produced N4,5-cyclic amide-2'-deoxycytidine containing both 14N and 15N atoms from uric acid-1,3-15N2. Singlet oxygen, hydroxyl radical, peroxynitrous acid, hypochlorous acid, and hypobromous acid generated neither N4-hydroxy-2'-deoxycytidine nor N4,5-cyclic amide-2'-deoxycytidine in the presence of uric acid. These results indicate that uric acid is a photosensitizer for the reaction of nucleosides by UV light with a wavelength longer than 300 nm, and that an unidentified radical derived from uric acid with a delocalized unpaired electron may be generated.

Advances in the phase separation-organized membraneless organelles in cells: a narrative review.

Membraneless organelles (MLOs) are micro-compartments that lack delimiting membranes, concentrating several macro-molecules with a high local concentration in eukaryotic cells. Recent studies have shown that MLOs have pivotal roles in multiple biological processes, including gene transcription, RNA metabolism, translation, protein modification, and signal transduction. These biological processes in cells have essential functions in many diseases, such as cancer, neurodegenerative diseases, and virus-related diseases. The liquid-liquid phase separation (LLPS) microenvironment within cells is thought to be the driving force for initiating the formation of micro-compartments with a liquid-like property, becoming an important organizing principle for MLOs to mediate organism responses. In this review, we comprehensively elucidated the formation of these MLOs and the relationship between biological functions and associated diseases. The mechanisms underlying the influence of protein concentration and valency on phase separation in cells are also discussed. MLOs undergoing the LLPS process have diverse functions, including stimulation of some adaptive and reversible responses to alter the transcriptional or translational processes, regulation of the concentrations of biomolecules in living cells, and maintenance of cell morphogenesis. Finally, we highlight that the development of this field could pave the way for developing novel therapeutic strategies for the treatment of LLPS-related diseases based on the understanding of phase separation in the coming years.

Application of Temporary, Cell-Containing Alginate Microcarriers to Facilitate the Fabrication of Spatially Defined Cell Pockets in 3D Collagen Hydrogels.

Mimicking the complexity of natural tissue is a major challenge in the field of tissue engineering. Here, a facile 2-step fabrication method to prepare 3D constructs with distinct regions of high cell concentrations and without the need for elaborate equipment is proposed. The initial incorporation of cells in a sacrificial alginate matrix allows the addition of other, cell relevant biopolymers, such as, collagen to form a spatially confined, interpenetrating network at the microscale. A layered structure at the macroscale can be achieved by incorporating these cell-containing microspheres in thin collagen layers. Cells are locally released by de-gelling the alginate matrix and their attachment to the collagen hydrogel layers has been studied. The use of the murine pre-osteoblast cell line MC3T3-E1 as an example cell line shows that the cells behave differently in their cell migration pattern based on the initial composition of the alginate microspheres.

Anaerobic granulation of single culture Clostridium beijerinckii

This study utilizes Clostridium beijerinckii P260, an anaerobic bacterium capable of acetone-butanol-ethanol (ABE) production, as model bacteria to test the hypothesis that single culture anaerobic granulation can be achieved in accordance with the biogranulation theory developed from the mixed culture studies. After four months of cultivation, C. beijerinckii granules formed under the selection pressure of a selective settling velocity at 9.0 m h−1 in a sequential batch reactor (SBR). These C. beijerinckii granules possess excellent settleability and exceptionally high cell and enzyme concentrations, which demonstrate promising potential in the application for high-rate fermentation and separation of value-added products. However, C. beijerinckii granules only accounted for 25% of the cell mass in the SBR, leaving the rest 75% still in the form of planktonic cells, which indicates an incomplete anaerobic granulation. The lack of ideal feast and famine conditions in the cyclic SBR operation was regarded as the reason. Future research should focus on the optimization of the bioreactor operation to create optimal conditions for inducing the phenotype switch from planktonic to granulated growth mode.

Cytotoxic and genotoxic evaluation of dipyridamole and its alternative therapeutic potential in cancer therapy: an in vitro and in vivo approach.

The molecular mechanism behind the anti-cancer potential of dipyridamole, an antiplatelet drug was investigated by examining the interaction of dipyridamole with DNA as well as the genotoxicity and cytotoxicity associated with dipyridamole . Interaction of dipyridamole with calf thymus DNA (Ct-DNA) was determined in order to ascertain the strength and mode of binding as well as the thermodynamic parameters involved. Dipyridamole interacts with Ct-DNA through a groove binding mechanism. Furthermore, a preliminary assessment of the genotoxic effect of dipyridamole was performed in vitro using the plasmid nicking assay and comet assay. Genotoxicity was also confirmed through DAPI nuclear staining of HeLa and HCT-116 cells. Cytotoxicity was assessed in vitro on HeLa, HCT-116 as well as HEK293 cells using the MTT assay as well as through inverted microscopy. Dipyridamole was selectively more toxic towards cancer cells. Dipyridamole demonstrated induction of micronucleus and chromosomal aberrations only at high concentrations in bone marrow cells of male Wistar rats, suggesting its safe usage at clinically relevant concentrations. These results indicated the existence of an intrinsic genotoxic potential of dipyridamole which may be contributing to its anti-cancer properties.

Properties of Degradable Polyhydroxyalkanoates (PHAs) Synthesized by a New Strain, Cupriavidus necator IBP/SFU-1, from Various Carbon Sources.

The bacterial strain isolated from soil was identified as Cupriavidus necator IBP/SFU-1 and investigated as a PHA producer. The strain was found to be able to grow and synthesize PHAs under autotrophic conditions and showed a broad organotrophic potential towards different carbon sources: sugars, glycerol, fatty acids, and plant oils. The highest cell concentrations (7–8 g/L) and PHA contents were produced from oleic acid (78%), fructose, glucose, and palm oil (over 80%). The type of the carbon source influenced the PHA chemical composition and properties: when grown on oleic acid, the strain synthesized the P(3HB-co-3HV) copolymer; on plant oils, the P(3HB-co-3HV-co-3HHx) terpolymer, and on the other substrates, the P(3HB) homopolymer. The type of the carbon source influenced molecular-weight properties of PHAs: P(3HB) synthesized under autotrophic growth conditions, from CO2, had the highest number-average (290 ± 15 kDa) and weight-average (850 ± 25 kDa) molecular weights and the lowest polydispersity (2.9 ± 0.2); polymers synthesized from organic carbon sources showed increased polydispersity and reduced molecular weight. The carbon source was not found to affect the degree of crystallinity and thermal properties of the PHAs. The type of the carbon source determined not only PHA composition and molecular weight but also surface microstructure and porosity of the polymer films. The new strain can be recommended as a promising P(3HB) producer from palm oil, oleic acid, and sugars (fructose and glucose) and as a producer of P(3HB-co-3HV) from oleic acid and P(3HB-co-3HV-co-3HHx) from palm oil.

Modeling and Optimization of Gas Sparging-Assisted Bacterial Cultivation Broth Microfiltration by Response Surface Methodology and Genetic Algorithm.

Production of highly efficient biomass-based microbial biopesticides significantly depends on downstream processing in terms of obtaining as high concentration of viable cells as possible. Microfiltration is one of the recommended operations for microbial biomass separation, but its main limitation is permeate flux decrease due to the membrane fouling. The effect of air sparging as a hydrodynamic technique for improvement of permeate flux during microfiltration of Bacillus velezensis cultivation broth was investigated. Modeling of the microfiltration was performed using the response surface methodology, while desirability function approach and genetic algorithm were applied for optimization, i.e., maximization of permeate flux and minimization of specific energy consumption. The results have revealed antagonistic relationship between the investigated dependent variables. The optimized values of superficial feed velocity and transmembrane pressure were close to the mean values of the investigated value ranges (0.68 bar and 0.96 m/s, respectively), while the optimized value of superficial air velocity had a more narrow distribution around 0.25 m/s. The results of this study have revealed a significant improvement of microfiltration performance by applying air sparging, thus this flux improvement method should be further investigated in downstream processing of different bacterial cultivation broths.

A New Method of Isolation of Mouse Renal Primary Tubular Epithelial Cells.

Kidney diseases are becoming an emerging public health problem. In order to further explore the etiology of various kidney diseases, we improved the methods of isolation of primary cultures of mouse renal tubular epithelial cells. At the first stage, the kidneys were perfused with collagenase solution. To this end, the superior mesenteric artery, celiac artery and thoracic aorta were ligated and perfusion was performed through the abdominal aorta. Then, the cells were isolated ex vivo and their integrity, purity, viability, and concentration were evaluated. The proposed cost-effective and simple method provides high purity and high concentration of primary renal epithelial cells for molecular biology studies of the kidneys.

O-148 Sperm Aminopeptidase N as a predictive biomarker of blastocyst development and embryo viability

Abstract Study question To evaluate human sperm APN as a prognostic factor for determining high-quality embryos. Summary answer The human sperm APN has the potential to become new molecular prognostic biomarker for having high-quality and viable embryos. What is known already Prognosis and diagnosis of male fertility is one of the major concerns in reproductive medicine. Approximately 30%-40% of men with otherwise normal fertility parameters are still unable to achieve pregnancy. The predictive clinical value of a semen analysis to identify fertile or infertile males is limited; therefore, new sperm diagnostic or prognostic methodologies are urgently required. Sperm Aminopeptidase N (APN) may be a relevant molecular marker due to its high concentration in sperm cells and its role in sperm physiology, such as motility, acrosome reaction, and embryo development. Study design, size, duration A prospective study that involves a total of 81 couples and 611 embryos who underwent oocyte-donation cycles at the Clínica IVI Bilbao (Spain) between September 2014 and July 2015. Participants/materials, setting, methods This study was set in an assisted reproduction unit and in an academic research laboratory. All semen samples were examined and classified following WHO guidelines. Spermatozoa were isolated from semen on discontinuous colloidal silica gradient (45%-90%) technique. Embryo quality and development were determined according to the Spanish Association of Reproduction Biology Studies (ASEBIR) criteria. Flow cytometry analyses of quantitative and semi-quantitative sperm human APN levels. Main results and the role of chance The obtained results proved that the most evolved and viable blastocysts were associated with low sperm APN levels. Expanding, expanded, hatching/hatched and viable blastocysts come from semen samples which showed lower APN levels than early blastocysts, blocked and non viable blastocyst. The cumulative probability of having more evolved blastyocysts increased 1.38-fold at day 5 and 1.98-fold at day 6 of embryo development as well as the likelihood of having viable embryo increased 1.48-fold when semen samples with low APN levels are used during the ICSI technique. Limitations, reasons for caution Data obtained from a single Fertility Clinic. A multi-centrum study will be required. Wider implications of the findings The human sperm APN has the potential to become new molecular prognostic biomarker for having high-quality embryos that could help to diagnose male infertility, especially when seminal parameters are close to the threshold values. Trial registration number Not applicable

Using qPCR and high-resolution sensor data to model a multi-species Pseudo-nitzschia (Bacillariophyceae) bloom in southeastern Australia

Harmful algal blooms, including those caused by the toxic diatom Pseudo-nitzschia, can have significant impacts on human health, ecosystem functioning and ultimately food security. In the current study we characterized a bloom of species of Pseudo-nitzschia that occurred in a south-eastern Australian oyster-growing estuary in 2019. Using light microscopy, combined with molecular (ITS/5.8S and LSU D1-D3 rDNA regions) and toxicological evidence, we observed the bloom to consist of multiple species of Pseudo-nitzschia including P. cf. cuspidata, P. hasleana, P. fraudulenta and P. multiseries, with P. cf. cuspidata being the only species that produced domoic acid (3.1 pg DA per cell). As several species of Pseudo-nitzschia co-occurred, only one of which produced DA, we developed a rapid, sensitive and efficient quantitative real-time polymerase chain reaction (qPCR) assay to detect only species belonging to the P. pseudodelicatissima complex Clade I, to which P. cf. cuspidata belongs, and this indicated that P. cuspidata or closely related strains may have dominated the Pseudo-nitzschia community at this time. Finally, using high resolution water temperature and salinity sensor data, we modeled the relationship between light microscopy determined abundance of P. delicatissima group and environmental variables (temperature, salinity, rainfall) at two sites within the estuary. A total of eight General Linear Models (GLMs) explaining between 9 and 54% of the deviance suggested that the temperature (increasing) and/or salinity (decreasing) data were generally more predictive of high cell concentrations than the rainfall data at both sites, and that overall, cell concentrations were more predictive at the more oceanic site than the more upstream site, using this method. We conclude that the combination of rapid molecular methods such as qPCR and real-time sensor data modeling, can provide a more rapid and effective early warning of harmful algal blooms of species of Pseudo-nitzschia, resulting in more beneficial regulatory and management outcomes.

An Integration Strategy to Develop Dual-State Luminophores with Tunable Spectra, Large Stokes Shift, and Activatable Fluorescence for High-Contrast Imaging

An Integration Strategy to Develop Dual-State Luminophores with Tunable Spectra, Large Stokes Shift, and Activatable Fluorescence for High-Contrast Imaging

Innovative fermented soya drink with the microalgae Chlorella vulgaris and the probiotic strain Levilactobacillus brevis ŁOCK 0944

The aim of this study is to produce a lactose-free product with Chlorella vulgaris, providing a large amount of lactic acid bacteria (LAB). The survival rate of Levilactobacillus brevis ŁOCK 0944 in soya drink with Chlorella vulgaris during 30 days of storage at 4 °C was higher than in soya drinks without algae. Moreover, Chlorella vulgaris caused an increase in l-lactic acid production by LAB in the soya drinks. From the high concentration of bacterial cells (8.49 log CFU/mL) in the fermented soya drink with algae after 30 days of storage at 4 °C, we can conclude that this microalga is a promising substrate for the production of fermented beverages. Furthermore, the soya drink with the Chlorella vulgaris and xylitol, due to the number of bacteria, also meets the criteria for a fermented probiotic product. The bacteria introduced into the gastrointestinal tract simulator together with a soya drink after the storage period exhibited a considerably higher survival rate, thus this indicating that the matrix provided protective conditions for them. The combination of Chlorella vulgaris and LAB in a soya drink provides the opportunity to create a functional plant-based product and opens new horizons for the food industry.

Effects of water stably-enriched with oxygen as a novel method of tissue oxygenation on mitochondrial function, and as adjuvant therapy for type 2 diabetes in a randomized placebo-controlled trial.

Diabetes mellitus is associated with inadequate delivery of oxygen to tissues. Cellular hypoxia is associated with mitochondrial dysfunction which increases oxidative stress and hyperglycaemia. Hyperbaric oxygenation therapy, which was shown to improve insulin sensitivity, is impractical for regular use. We evaluated the effects of water which is stably-enriched with oxygen (ELO water) to increase arterial blood oxygen levels, on mitochondrial function in the presence of normal- or high-glucose environments, and as glucose-lowering therapy in humans. We compared arterial blood oxygen levels in Sprague-Dawley rats after 7 days of ad libitum ELO or tap water consumption. Mitochondrial stress testing, and flow cytometry analysis of mitochondrial mass and membrane potential, were performed on human HepG2 cells cultured in four Dulbecco's Modified Eagle Medium media, made with ELO water or regular (control) water, at normal (5.5 mM) or high (25 mM) glucose concentrations. We also randomized 150 adults with type 2 diabetes (mean age 53 years, glycated haemoglobin HbA1c 8.9% [74 mmol/mol], average duration of diabetes 12 years) to drink 1.5 litres daily of bottled ELO water or drinking water. ELO water raised arterial oxygen tension pO2 significantly (335 ± 26 vs. 188 ± 18 mmHg, p = 0.006) compared with tap water. In cells cultured in control water, mitochondrial mass and membrane potential were both significantly lower at 25 mM glucose compared with 5.5 mM glucose; in contrast, mitochondrial mass and membrane potential did not differ significantly at normal or high glucose concentrations in cells cultured in ELO water. The high-glucose environment induced a greater mitochondrial proton leak in cells cultured in ELO water compared to cells cultured in control medium at similar glucose concentration. In type 2 diabetic adults, HbA1c decreased significantly (p = 0.002) by 0.3 ± 0.7% (4 ± 8 mmol/mol), with ELO water after 12 weeks of treatment but was unchanged with placebo. ELO water raises arterial blood oxygen levels, appears to have a protective effect on hyperglycaemia-induced reduction in mitochondrial mass and mitochondrial dysfunction, and may be effective adjuvant therapy for type 2 diabetes.