Reduction In Intracellular pH Research Articles

Combination treatment with histone deacetylase and carbonic anhydrase 9 inhibitors shows therapeutic potential in experimental diffuse intrinsic pontine glioma.

Diffuse intrinsic pontine glioma (DIPG) remains a significant therapeutic challenge due to the lack of effective and safe treatment options. This study explores the potential of combining histone deacetylase (HDAC) and carbonic anhydrase 9 (CA9) inhibitors in treating DIPG. Analysis of RNA sequencing data and tumor tissue from patient samples for the expression of the carbonic anhydrase family and hypoxia signaling pathway activity revealed clinical relevance for targeting CA9 in DIPG. A synergy screen was conducted using CA9 inhibitor SLC-0111 and HDAC inhibitors panobinostat, vorinostat, entinostat, and pyroxamide. The combination of SLC-0111 and pyroxamide demonstrated the highest synergy and was selected for further analysis. Combining SLC-0111 and pyroxamide effectively inhibited DIPG cell proliferation, reduced cell migration and invasion potential, and enhanced histone acetylation, leading to decreased cell population in S Phase. Additionally, the combination therapy induced a greater reduction in intracellular pH than either agent alone. Data from this study suggest that the combination of SLC-0111 and pyroxamide holds promise for treating experimental DIPG, and further investigation of this combination therapy in preclinical models is warranted to evaluate its potential as a viable treatment for DIPG.

Antibacterial mechanism of CO2 combined with low temperature against Shewanella putrefaciens by biochemical and metabolomics analysis

To further reveal the inhibition mechanism of carbon dioxide (CO2) on Shewanella putrefaciens (S. putrefaciens), influence on metabolic function was studied by biochemical and metabolomics analysis. Accordingly, reduction of intracellular pH (pHi), depolarization of cell membrane and accumulation of reactive oxygen species (ROS) indicated that CO2 changed the membrane permeability of S. putrefaciens. Besides, adenosine triphosphate (ATP), ATPase, nicotinamide adenine dinucleotide (NAD+/NADH) and ratios of NADH/NAD+ were detected, indicating a role of CO2 in repressing respiratory pathway and electron transport. According to metabolomics results, CO2 induced differential expressions of metabolites, disordered respiratory chain and weakened energy metabolism of S. putrefaciens. Inhibition of respiratory rate-limiting enzymes also revealed that electron transfer of respiratory chain was blocked, cell respiration was weakened, and thus energy supply was insufficient under CO2 stress. These results revealed that CO2 caused disruption of metabolic function, which might be the main cause of growth inhibition for S. putrefaciens.

Fisetin-Mediated Perturbations of Membrane Permeability and Intracellular pH in Candida albicans.

The antifungal activity of fisetin against Candida albicans is explored, elucidating a mechanism centered on membrane permeabilization and ensuing disruption of pH homeostasis. The Minimum Inhibitory Concentration (MIC) of fisetin, indicative of its interaction with the fungal membrane, increases in the presence of ergosterol. Hoechst 33342 and propidium-iodide staining reveal substantial propidium-iodide accumulation in fisetin-treated C. albicans cells at their MIC, with crystal violet uptake assays confirming fisetin-induced membrane permeabilization. Leakage analysis demonstrates a significant release of DNA and proteins in fisetin-treated cells compared to controls, underscoring the antifungal effect through membrane disruption. Green fluorescence, evident in both the cytoplasm and vacuoles of fisetin-treated cells under BCECF, AM staining, stands in contrast to controls where only acidic vacuoles exhibit staining. Ratiometric pH measurements using BCECF, AM reveal a noteworthy reduction in intracellular pH in fisetin-treated cells, emphasizing its impact on pH homeostasis. DiBAC4(3) uptake assays demonstrate membrane hyperpolarization in fisetin-treated cells, suggesting potential disruptions in ion flux and cellular homeostasis. These results provide comprehensive insights into the antifungal mechanisms of fisetin, positioning it as a promising therapeutic agent against Candida infections.

Antibacterial mechanism of Protocatechuic acid against Yersinia enterocolitica and its application in pork

Protocatechuic acid (PCA) has adequate antibacterial activity against foodborne pathogens; however, its antibacterial mechanism has been rarely investigated. Therefore, this study attempts to elucidate the antibacterial mechanism of PCA against Yersinia enterocolitica . The corresponding results showed that PCA had a satisfactory antibacterial effect on Y. enterocolitica with a minimum inhibitory concentration (MIC) of 2.5 mg/mL. Accordingly, depolarization of the cell membrane, reduction of intracellular pH (pH in ) and adenosine triphosphate (ATP), leakage of K + and destruction of cell morphology were observed in this study, which indicated that PCA changed the membrane permeability of Y. enterocolitica . According to the metabolomics analysis, PCA induced the differential expression of the metabolites of Y. enterocolitica . In addition, enrichment analysis of metabolic pathways demonstrated that PCA disordered energy metabolism and amino acid biosynthesis of Y. enterocolitica . PCA also exhibited a notable inhibitory effect on Y. enterocolitica in pork. According to the corresponding findings, PCA may possess potential antibacterial properties in controlling Y. enterocolitica contamination within food industry. • Protocatechuic acid showed high antibacterial activity against Y. enterocolitica. • Protocatechuic acid destroyed the cell membrane structure of Y. enterocolitica. • Protocatechuic acid disordered energy metabolism and amino acid metabolism. • Protocatechuic acid extended the shelf life of chilled pork.

Phenotypic and Transcriptomic Analyses Reveal the Cell Membrane Damage of Pseudomonas fragi Induced by Cinnamic Acid.

Cinnamic acid (CA) is a safe and effective antimicrobial agent. The objective of this study was to reveal the antibacterial mechanism of CA against a food-derived Pseudomonas fragi 38-8, from the aspects of bacterial growth kinetics, cell membrane homeostasis, cell microstructure, and transcription. The minimum inhibitory concentration (MIC) of CA against P. fragi 38-8 was 0.25 mg/ml. CA retarded bacterial growth and induced a series of cell membrane changes. After CA treatment, cell membrane homeostasis was destroyed, which was evidenced by cell membrane depolarization, intracellular pH reduction, and intracellular ATPase activity decrease. Field emission scanning electron microscope (FESEM), transmission electron microscope (TEM), and confocal laser scanning fluorescence microscope (CLSM) realized the visualization of cell microstructure changes, showing cell death and morphological changes, such as cell rupture, shrinkage, and hollowness. RNA sequencing analysis further confirmed the effects of CA to the cell membrane, because of the significant enrichment of differentially expressed genes (DEGs) related to membrane. The results of the phenotype tests and RNA-seq both focused on cell membrane damage, which showed that CA exerted antibacterial effect mainly by acting on cell membrane.

Yeast stress and death caused by the synergistic effect of ethanol and SO<SUB>2</SUB> during the second fermentation of sparkling wines

Problems can often arise at the beginning of the second fermentation (prise de mousse) of sparkling wines, such as no start, a long lag period or slow fermentation. These problems are generally associated with yeast stress when inoculated in a base wine with high ethanol content and low pH. However, few studies focus on sulphites, which are often added to base wines to prevent malolactic fermentation, microbiological instability, and wine oxidation. This study aimed to evaluate the joint effect of ethanol and sulfur dioxide on yeasts during the second fermentation. For this purpose, yeasts (Saccharomyces cerevisiae EC1118) were subjected to ethanol, sulfur dioxide and ethanol/sulfur dioxide at the beginning of fermentation, and their vitality and viability, as well as the accumulation of intracellular reactive oxygen species and intracellular pH, were evaluated by flow cytometry. Furthermore, the expression of genes involved in sulfur transport and metabolism was determined. The results showed high mortality, ROS accumulation and intracellular pH reduction in fermentations with both ethanol and sulfur dioxide. The negative effect of ethanol, sulfur dioxide and ethanol/sulfur dioxide on yeasts was found to be dose-dependent and high in those commonly found in some base wines. Cells treated with ethanol/sulfur dioxide showed over-expression of genes involved in sulphite transport (SUL1 and SUL2), efflux pump (SSU1 and FZF1) and metabolism of sulfur amino acids (MET14). Altogether, our data indicate that ethanol and sulfur dioxide have a synergistic effect on yeasts, which may be the root cause of the problems encountered at the beginning of the second fermentation of sparkling wines, and should thus be seriously taken into consideration by winemakers.

Physicochemical and transcriptomic responses of Lactobacillus brevis JLD715 to sodium selenite.

Elemental selenium, as a new type of selenium supplement, can be prepared by microorganisms reducing inorganic selenium. In this study, Lactobacillus brevis JLD715 was incubated in broth containing different concentrations of sodium selenite (Na2 SeO3 ). The results showed that the bacterial biomass of L. brevis JLD715 decreased due to the inhibition of Na2 SeO3 . The cell membrane of L. brevis JLD715 treated with Na2 SeO3 was damaged, as evidenced by the reduction of intracellular ATP concentration, depolarization of cell membrane, reduction of intracellular pH and impairment of membrane integrity. In addition, we investigated the metabolism mechanism of Na2 SeO3 by L. brevis JLD715 based on transcriptome sequencing. A total of 461 genes were significantly differentially expressed under Na2 SeO3 treatment, of which 231 genes were up-regulated and 230 genes were down-regulated. These genes were involved in pathways such as pyruvate metabolism, fatty acid biosynthesis, selenocompound metabolism and nucleotide-binding oligomerization domain-like (NOD-like) receptor signaling. Meanwhile, the genes related to sulfhydryl oxidoreductase, electron carrier proteins and transmembrane transport proteins synthesis were significantly up-regulated. To sum up, the findings of this research will contribute to providing support for the application of L. brevis JLD715 in selenium-enriched functional foods. © 2021 Society of Chemical Industry.

Oxaliplatin-induced neuropathy occurs through impairment of haemoglobin proton buffering and is reversed by carbonic anhydrase inhibitors.

Oxaliplatin is a cornerstone chemotherapeutic used in the treatment of colorectal cancer, the third leading cause of death in Western countries. Most side effects of this platinum-containing drug are adequately managed in the clinic, although acute and long-term neurotoxicity still severely compromises the quality of life of patients treated with oxaliplatin. We have previously demonstrated that therapeutically relevant concentrations/doses of oxaliplatin lead to a reduction in intracellular pH in mouse dorsal root ganglion (DRG) neurons in vitro and in vivo and that this alteration sensitizes TRPA1 and TRPV1 channels, which most likely mediate the allodynia associated with treatment. In this study, we show that oxaliplatin leads to a reduction of intracellular pH by forming adducts with neuronal haemoglobin, which acts in this setting as a proton buffer. Furthermore, we show that FDA-approved drugs that inhibit carbonic anhydrase (an enzyme that is linked to haemoglobin in intracellular pH homeostasis), ie, topiramate and acetazolamide, revert (1) oxaliplatin-induced cytosolic acidification and TRPA1 and TRPV1 modulation in DRG neurons in culture, (2) oxaliplatin-induced cytosolic acidification of DRG of treated animals, and (3) oxaliplatin-induced acute cold allodynia in mice while not affecting OHP-induced cytotoxicity on cancer cells. Our data would therefore suggest that reversal of oxaliplatin-induced cytosolic acidification is a viable strategy to minimize acute oxaliplatin-induced symptoms.

Curcumin Treatment in Combination with Glucose Restriction Inhibits Intracellular Alkalinization and Tumor Growth in Hepatoma Cells.

Dysregulation of cellular energy metabolism is closely linked to cancer development and progression. Calorie or glucose restriction (CR or GR) inhibits energy-dependent pathways, including IGF-1/PI3K/Akt/mTOR, in cancer cells. However, alterations in proton dynamics and reversal of the pH gradient across the cell membrane, which results in intracellular alkalinization and extracellular acidification in cancer tissues, have emerged as important etiopathogenic factors. We measured glucose, lactate, and ATP production after GR, plant-derived CR-mimetic curcumin treatment, and curcumin plus GR in human hepatoma cells. Intracellular pH regulatory effects, in particular, protein–protein interactions within mTOR complex-1 and its structural change, were investigated. Curcumin treatment or GR mildly inhibited Na+/H+ exchanger-1 (NHE1). vATPase, monocarboxylate transporter (MCT)-1, and MCT4 level. Combination treatment with curcumin and GR further enhanced the inhibitory effects on these transporters and proton-extruding enzymes, with intracellular pH reduction. ATP and lactate production decreased according to pH change. Modeling of mTOR protein revealed structural changes upon treatments, and curcumin plus GR decreased binding of Raptor and GβL to mTOR, as well as of Rag A and Rag B to Raptor. Consequently, 4EBP1 phosphorylation was decreased and cell migration and proliferation were inhibited in a pH-dependent manner. Autophagy was increased by curcumin plus GR. In conclusion, curcumin treatment combined with GR may be a useful supportive approach for preventing intracellular alkalinization and cancer progression.

A Preliminary Computational Model for Hypoxic Acidosis in Cardiac Myocytes.

Intracellular acidosis induced by hypoxia resulted from myocardial ischemia damages myocardium. However, the detailed mechanisms of hypoxic acidosis are not quantitatively explained. The purpose of this study is to create a novel computational model which can reproduce intracellular acidosis caused by myocardial ischemia. We constructed a computational model of myocardium, by using a mathematical ventricular cell model which includes pH regulation and a computational model of myocardial microcirculation for calculating extracellular conditions. The present model reproduced cellular hypoxia in an ischemic condition simulated by a reduced blood flow, and intracellular pH reduction in response to the hypoxia.

Escherichia coli inactivation by pressurized CO2 treatment methods at room temperature: Critical issues

This study aims to increase the inactivation efficiency of CO2 against Escherichia coli under mild conditions to facilitate the application of pressurized CO2 technology in water disinfection. Based on an aerating-cycling apparatus, three different treatment methods (continuous aeration, continuous reflux, and simultaneous aeration and reflux) were compared for the same temperature, pressure (0.3–0.7MPa), initial concentration, and exposure time (25min). The simultaneous aeration and reflux treatment (combined method) was shown to be the best method under optimum conditions, which were determined to be 0.7MPa, room temperature, and an exposure time of 10min. This treatment achieved 5.1-log reduction after 25min of treatment at the pressure of 0.3MPa and 5.73-log reduction after 10min at 0.7MPa. Log reductions of 4.4 and 5.0 occurred at the end of continuous aeration and continuous reflux treatments at 0.7MPa, respectively. Scanning electron microscopy (SEM) images suggested that cells were ruptured after the simultaneous aeration and reflux treatment and the continuous reflux treatment. The increase of the solubilization rate of CO2 due to intense hydraulic conditions led to a rapid inactivation effect. It was found that the reduction of intracellular pH caused by CO2 led to a more lethal bactericidal effect.

Widespread pH abnormalities in patients with malformations of cortical development and epilepsy: A phosphorus-31 brain MR spectroscopy study

Neuroimaging studies demonstrate that not only the lesions of malformations of cortical development (MCD) but also the normal-appearing parenchyma (NAP) present metabolic impairments, as revealed with (1)H-MRS. We have previously detected biochemical disturbances in MCD lesions with phosphorus-31 magnetic resonance spectroscopy (31P-MRS). Our hypothesis is that pH abnormalities extend beyond the visible lesions. Three-dimensional 31P-MRS at 3.0 T was performed in 37 patients with epilepsy and MCD, and in 31 matched-control subjects. The patients were assigned into three main MCD subgroups: cortical dysplasia (n=10); heterotopia (n=14); schizencephaly/polymicrogyria (n=13). Voxels (12.5 cm3) were selected in five homologous regions containing NAP: right putamen; left putamen; frontoparietal parasagittal cortex; right centrum semiovale; and left centrum semiovale. Robust methods of quantification were applied, and the intracellular pH was calculated with the chemical shifts of inorganic phosphate (Pi) relative to phosphocreatine (PCr). In comparison to controls and considering a Bonferroni adjusted p-value <0.01, MCD patients presented significant reduction in intracellular pH in the frontoparietal parasagittal cortex (6.985±0.022), right centrum semiovale (7.004±0.029), and left centrum semiovale (6.995±0.030), compared to controls (mean values±standard deviations of 7.087±0.048, 7.096±0.042, 7.088±0.045, respectively). Dunnet and Dunn tests demonstrated that the differences in pH values remained statistically significant in all MCD subgroups. No significant differences were found for the putamina. The present study demonstrates widespread acidosis in the NAP, and reinforces the idea that MCD visible lesions are only the tip of the iceberg.

Similarities between Exogenously- and Endogenously-Induced Envelope Stress: The Effects of a New Antibacterial Molecule, TPI1609-10

Antibiotics with novel and/or multiple targets are highly desirable in the face of the steady rise of clinical antibiotic resistance. We have screened and identified small molecules, typified by the compound TPI1609-10 (aka SM10), with antibiotic activity against both gram-positive and gram-negative bacteria. SM10 was screened in vitro to bind branched Holliday junction intermediates of homologous recombination and tyrosine recombinase-mediated recombination; thus, the cellular targets of the small molecules were expected to include the RuvABC Holliday junction resolvasome and the XerCD complex involved in proper segregation of replicated chromosomes to daughter cells. SM10 indeed induces DNA damage and filamentation in E. coli. However, SM10 also induces envelope stress and causes increased production of intracellular reactive oxygen species. In addition, SM10 has similar effects to endogenously-induced envelope stress via overproducing outer membrane proteins (OmpC and OmpF), which also induces the SOS response, chromosome fragmentation, and production of reactive oxygen species. The synergy between SM10, and cerulenin, a fatty acid synthesis inhibitor, together with the SM10 hypersensitivity of cpx and rpoE mutants, further support that SM10's mode of action damages membrane damage. The lethality of SM10 treatment and of OmpC overproduction are observed in both aerobically- and anaerobically-grown cells, and is accompanied by substantial DNA damage even anaerobically. Thus, only some DNA damage is due to reactive oxygen. We propose that membrane depolarization and the potential reduction in intracellular pH, leading to abasic site formation, cause a substantial amount of the DNA damage associated with both SM10 treatment and endogenous envelope stress. While it is difficult to completely exclude effects related to envelope damage as the sources of DNA damage, trapping intermediates associated with DNA repair and chromosome segregation pathways remains very likely. Thus SM10 may have distinct but synergistic modes of action.

β-alanine supplementation improves YoYo intermittent recovery test performance

Backgroundβ-alanine supplementation has been shown to improve high-intensity exercise performance and capacity. However, the effects on intermittent exercise are less clear, with no effect shown on repeated sprint activity. The aim of this study was to investigate the effects of β-alanine supplementation on YoYo Intermittent Recovery Test Level 2 (YoYo IR2) performance.MethodsSeventeen amateur footballers were allocated to either a placebo (PLA; N = 8) or β-alanine (BA; N = 9) supplementation group, and performed the YoYo IR2 on two separate occasions, pre and post 12 weeks of supplementation during a competitive season. Specifically, players were supplemented from early to mid-season (PLA: N = 5; BA: N = 6) or mid- to the end of the season (PLA: N = 3; BA: N = 3). Data were analysed using a two factor ANOVA with Tukey post-hoc analyses.ResultsPre supplementation scores were 1185 ± 216 and 1093 ± 148 m for PLA and BA, with no differences between groups (P = 0.41). YoYo performance was significantly improved for BA (+34.3%, P ≤ 0.001) but not PLA (−7.3%, P = 0.24) following supplementation. 2 of 8 (Early – Mid: 2 of 5; Mid – End: 0 of 3) players improved their YoYo scores in PLA (Range: -37.5 to + 14.7%) and 8 of 9 (Early – Mid: 6 of 6; Mid – End: 2 of 3) improved for BA (Range: +0.0 to +72.7%).Conclusions12 weeks of β-alanine supplementation improved YoYo IR2 performance, likely due to an increased muscle buffering capacity resulting in an attenuation of the reduction in intracellular pH during high-intensity intermittent exercise.

Process analysis of reduced specific productivity of TNFR-Fc in Chinese hamster ovary cells at high cell density

Specific TNFR-Fc productivity ( q TNFR-Fc) in CHO cells was found to decrease by 68% at a cell density of 6 × 10 6 cells/mL compared to that at 2 × 10 6 cells/mL. In order to unravel the reasons for the reduced q TNFR-Fc at high cell density, an in-depth study of physiological factors, gene transcription and post-translational process was carried out. Physiological factors including cell biomass, intracellular protein and cell cycle distribution were not affected by cell density. Transcriptional analysis revealed that TNFR-Fc mRNA level at 2 × 10 6 cells/mL was almost 2-fold higher than that at 6 × 10 6 cells/mL. Moreover, rather than mRNA stability, the slower transcriptional rate led to the decreased mRNA level. By analyzing the post-translational process, we found that the protein assembly rate and transport rate from the endoplasmic reticulum to the Golgi at high cell density were significantly lower than those at low cell density. Therefore, the decreased q TNFR-Fc at high cell density was correlated with the reduction in both mRNA level and post-translational processing rate. It was further found that the decreased mRNA level and post-translational processing rate might be linked to the apparent increase in CO 2 partial pressure (pCO 2) at high cell density, which had resulted in the reduction in intracellular pH in CHO cells.

Lactateper seimproves the excitability of depolarized rat skeletal muscle by reducing the Cl−conductance

Studies on rats have shown that lactic acid can improve excitability and function of depolarized muscles. The effect has been related to the ensuing reduction in intracellular pH causing inhibition of muscle fibre Cl(-) channels. However, since several carboxylic acids with structural similarities to lactate can inhibit muscle Cl(-) channels it is possible that lactate per se can increase muscle excitability by exerting a direct effect on these channels. We therefore examined the effects of lactate on the function of intact muscles and skinned fibres together with effects on pH and Cl(-) conductance (G(cl)). In muscles where extracellular compound action potentials (M-waves) and tetanic force response to excitation were reduced by (mean ± s.e.m.) 82 ± 4% and 83 ± 2%, respectively, by depolarization with 11 mm extracellular K(+), both M-waves and force exhibited an up to 4-fold increase when 20 mm lactate was added. This effect was present already at 5 mm and saturated at 15 mm lactate, and was associated with a 31% reduction in G(Cl). The effects of lactate were completely blocked by Cl(-) channel inhibition or use of Cl(-)-free solutions. Finally, both experiments where effects of lactate on intracellular pH in intact muscles were mimicked by increased CO₂ tension and experiments with skinned fibres showed that the effects of lactate could not be related to reduced intracellular pH. It is concluded that addition of lactate can inhibit ClC-1 Cl(-) channels and increase the excitability and contractile function of depolarized rat muscles via mechanisms not related to a reduction in intracellular pH.

NHE1, NHE3, and NHE5 mRNA expression in brainstem and cortex of Sprague‐Dawley rat

In brainstem CO2 chemosensitive neurons, an increase in CO2 (hypercapnia) leads to a maintained reduction in intracellular pH (pHi) while in non‐chemosensitive neurons, pHi recovery is observed. Although the precise mechanisms for this difference in pHi regulation remain to be determined, it has been suggested that different isoforms of the Na+/H+‐exchanger (NHE) may be responsible. To begin to explore this possibility, real‐time RT‐PCR was used to examine mRNA expression for three NHE isoforms ‐ NHE1 (Slc9a1), NHE3 (Slc9a3), and NHE5 (Slc9a5) ‐ in brainstem and cortex (non‐chemosensitive control) of 21 day‐old SD rat. Preliminary data indicate that each of the NHE isoforms is present in both brainstem and cortex. To further investigate the distribution within the medulla, mRNA expression levels for the NHE3 and NHE5 in dorsal (dM) versus ventral (vM) medulla were compared. These analyses revealed that NHE5 mRNA levels are ~4‐fold higher in the vM than in the dM (when normalized to Actb), but NHE3 mRNA levels are similar between the two regions. Additional studies are needed to further evaluate the relative abundance of different NHE mRNAs in chemosensitive areas such as the NTS/DMV, VLM, the caudal medullary raphe, and the LC. To date, however, our data suggest that differential expression of NHE isoforms may participate in pHi regulation in chemosensitive and/or non‐chemosensitive neurons. Supported by NS045321.

Low luminal pH aggravates fatty acid bound albumin induced O2·− production in renal proximal tubular cell

It is known that fatty acid bound albumin induces oxidative stress in proximal tubular cells. Although luminal pH of proximal tubule decreases according to the reabsorption of bicarbonate, it is not examined well how luminal pH affects the production of superoxide, O2·−. Therefore, we examined the role of luminal pH in the O2·− production.We applied oleic acid bound albumin(OA‐alb, 15g/L) to the HK‐2 cells, human proximal tubule cell line, and measured the production with dihydroethidium. The production was evaluated as the increasing rate of ethidium intensity under real time fluorescent microscopy. When buffer pH is above 6.6, the production was not significant(O2·− production; 0.15 ± 0.06 unit/sec at pH 6.9 and 0.34 ± 0.15 unit/sec at pH6.6, p=0.297). However, by decreasing buffer pH to 6.4, OA‐alb increased the production to 1.45 ± 0.28 unit/sec (p=0.026 versus pH 6.6). A Na/H exchanger inhibitor, dimethylamiloride blunted the production by 62%.In summary, the luminal acidic pH, which activates Na/H exchanger through the reduction of intracellular pH, aggravates OA‐alb induced O2·− production in proximal tubular cells.

A mathematical model of pH i regulation in central CO 2 chemoreception

In CO2 chemosensitive neurons, an increase in CO2 (hypercapnia) leads to a maintained reduction in intracellular pH (pHi) while in non-chemosensitive neurons pHi recovery is observed. The precise mechanisms for the differential regulation of pHi recovery between these cell populations remain to be identified; however, studies have begun to explore the role of Na+/H+ exchange (NHE). Here we present the results of a mathematical model that explores pHi regulation in central CO, chemoreception, and in particular the role of the isoform NHE-5 (present in brainstem neurons). We found that NHE-5 kinetics leads to pHi recovery in response to simulated hypercapnia as we already reported for the isoforms NHE-1 and NHE-3. Further, we wanted to test whether our model can reproduce common experimental protocols like NH4 + pre-pulse experiments designed to alter pHi.

Exploiting hypoxia in solid tumors to achieve oncolysis

Chemo- and radio-resistant cancer cells within solid tumors undermine the effectiveness of these approaches to achieving oncolysis. These resistant cells and clusters of cells typically thrive at low oxygen tensions and are reliant on anaerobic metabolic pathways that churn out lactate. This hypoxic state is one that can be exploited and in this paper a novel method is advanced involving tumor cell infiltration by bifidobacterium species which should bring about prodigious lactate synthesis; concomitant blocking of its enzymatic degradation by urea as well as export (from the cell) by use of quercetin; depletion of ATP using exogenous thyroid; and compromised oxidative catabolism of free fatty acids and amino acids via oral intake of l-hydroxycitrate, melatonin and nontoxic NDGA. This "anaerobic pathway cocktail", it is hypothesized, will bring about a profound reduction in intracellular pH and a compromised state of cellular energetics sufficient to effect oncolysis.