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Biopreservation and Biobanking Jun 2022The safety of banked human adipose-derived stem cells (hADSCs) purified by 155 mM ammonium chloride (NHCl)-based erythrocyte lysis has not been evaluated. This study...
The safety of banked human adipose-derived stem cells (hADSCs) purified by 155 mM ammonium chloride (NHCl)-based erythrocyte lysis has not been evaluated. This study was conducted to determine the impact of NHCl-based erythrocyte lysis on the biological characteristics of cryopreserved hADSCs. Stromal vascular fractions (SVFs) were obtained from lipoaspirates and purified with NHCl-based erythrocyte lysis (lysis group) or without (nonlysis group). The hADSCs were freshly isolated (fresh group) from SVFs and/or cryopreserved for 2 weeks (cryo group). The morphologies, immunophenotypes, viability, apoptosis, and growth kinetics of each group were compared. The cell cycle and differentiation capacity assays were performed in both cryopreserved groups. All groups showed similar cell morphology, immunological phenotypes, and viability. However, the main effect of lysis and its interaction with cryopreservation were observed when early apoptosis was regarded as a dependent variable in two-way repeated-measures analysis of variance. After cryopreservation, significant growth retardation and S-phase fraction reduction were observed in lytic hADSCs compared with those in nonlytic hADSCs. No significant differences in the adipogenic and osteogenic differentiation capacities were found between the two groups. Although NHCl-based erythrocyte lysis did not affect the cell morphology, immunological phenotypes, viability, and adipogenic and osteogenic differentiation capacities of cryopreserved hADSCs, exposure to NHCl-based erythrocyte lysis or its synergistic action with cryopreservation may induce apoptosis and inhibit the proliferation and mitosis of cryopreserved hADSCs. These results indicate that NHCl-based erythrocyte lysis is not suitable for high-quality banked collection of hADSCs for future clinical applications. Further development of safe, convenient, and cost-effective purification methods of hADSCs is warranted.
Topics: Adipose Tissue; Ammonium Chloride; Cell Differentiation; Cells, Cultured; Erythrocytes; Osteogenesis; Stem Cells
PubMed: 34704812
DOI: 10.1089/bio.2021.0035 -
Materials (Basel, Switzerland) Nov 2019Benzyl ammonium chloride (BAC) is a broad-spectrum bactericide, but vulnerable to leaching by water. In this paper, halloysite nanotubes (HNT) and montmorillonite (MMT)...
Benzyl ammonium chloride (BAC) is a broad-spectrum bactericide, but vulnerable to leaching by water. In this paper, halloysite nanotubes (HNT) and montmorillonite (MMT) were used as drug carriers to load BAC, in order to achieve good anti-mildew activity and long-term sustained release properties. The HNT and MMT nano-composites were characterized by transmission electron microscope (TEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), and nitrogen adsorption/desorption. XRD results showed that BAC intercalated the interlayer of MMT, and expanded the interlayer spacing from 1.15 nm to 1.75 nm. Pore analysis showed that BAC decreased the cavity of halloysite nanotubes to a certain extent, which indicated that BAC loaded inside the lumen of HNT successfully. TG analysis showed that the loading capacity of MMT to BAC was higher than HNT. The accelerated-release experiments revealed both two clays have significant sustained-release effects on BAC, and the releasing rate of HNT was relatively lower. Both HNT and MMT have promising application prospects as sustained-release carriers. The inhibition test showed that BAC in nano-clay has good anti-mildew resistance performance.
PubMed: 31752088
DOI: 10.3390/ma12223780 -
Experimental and Therapeutic Medicine Jul 2024Ammonia (NH) is an irritating and harmful gas that affects cell apoptosis and autophagy. Sirtuin 5 (SIRT5) has multiple enzymatic activities and regulates NH-induced...
Ammonia (NH) is an irritating and harmful gas that affects cell apoptosis and autophagy. Sirtuin 5 (SIRT5) has multiple enzymatic activities and regulates NH-induced autophagy in tumor cells. In order to determine whether SIRT5 regulates NH-induced bovine mammary epithelial cell apoptosis and autophagy, cells with SIRT5 overexpression or knockdown were generated and in addition, bovine mammary epithelial cells were treated with SIRT5 inhibitors. The results showed that SIRT5 overexpression reduced the content of NH and glutamate in cells by inhibiting glutaminase activity in glutamine metabolism, and reduced the ratio of ADP/ATP. The results in the SIRT5 knockdown and inhibitor groups were comparable, including increased content of NH and glutamate in cells by activating glutaminase activity, and an elevated ratio of ADP/ATP. It was further confirmed that SIRT5 inhibited the apoptosis and autophagy of bovine mammary epithelial cells through reverse transcription-quantitative PCR, western blot, flow cytometry with Annexin V FITC/PI staining and transmission electron microscopy. In addition, it was also found that the addition of LY294002 or Rapamycin inhibited the PI3K/Akt or mTOR kinase signal, decreasing the apoptosis and autophagy activities of bovine mammary epithelial cells induced by SIRT5-inhibited NH. In summary, the PI3K/Akt/mTOR signal involved in NH-induced cell autophagy and apoptosis relies on the regulation of SIRT5. This study provides a new theory for the use of NH to regulate bovine mammary epithelial cell apoptosis and autophagy, and provides guidance for improving the health and production performance of dairy cows.
PubMed: 38827477
DOI: 10.3892/etm.2024.12584 -
PloS One 2023Bone disease is highly prevalent in patients with chronic kidney disease (CKD), leading to an increased risk of bone fractures. This is due in part to metabolic...
Bone disease is highly prevalent in patients with chronic kidney disease (CKD), leading to an increased risk of bone fractures. This is due in part to metabolic acid-induced bone dissolution. Bisphosphonates (BPPs) are a potential treatment for inhibiting bone dissolution; however, there are limited studies observing the use of BPPs on acidotic patients. We aimed to determine efficacy of BPPs on maintaining bone health and pH regulation in acid-exposed mice. Using a diet-induced murine model of metabolic acidosis, we examined bone structure, composition, and mechanics as well as blood gases for three groups: control, acidosis, and acidosis + bisphosphonates (acidosis+BPP). Acidosis was induced for 14 days and alendronate was administered every 3 days for the acidosis+BPP group. The administration of BPP had little to no effect on bone structure, mechanics, and composition of the acidosis bones. However, administration of BPP did cause the mice to develop more severe acidosis than the acidosis only group. Overall, we discovered that BPPs may exacerbate acidosis symptoms by inhibiting the release of buffering ions from bone. Therefore, we propose that BPP administration should be carefully considered for those with CKD and that alkali supplementation could help minimize acidifying effects.
Topics: Animals; Mice; Alendronate; Ammonium Chloride; Diphosphonates; Acidosis; Osteolysis; Renal Insufficiency, Chronic
PubMed: 37713420
DOI: 10.1371/journal.pone.0291649 -
Renal Failure Dec 2022To examine the dynamic changes in the formative factors of nephrolithiasis and the final micromorphological changes in an obesity-initiated metabolic syndrome (MS) rat...
INTRODUCTION AND OBJECTIVE
To examine the dynamic changes in the formative factors of nephrolithiasis and the final micromorphological changes in an obesity-initiated metabolic syndrome (MS) rat model.
METHODS
Forty five-week-old male Sprague-Dawley (SD) rats were randomly divided into four groups: the regular diet group (RD), high-fat diet group (HFD), regular diet with drug (ethylene glycol and ammonium chloride) group (RDD), and high-fat diet with drug group (HFDD). A dynamic assessment of MS components (body weight (BW), body length (BL), Lee's index (LI), blood glucose (BG), total cholesterol (TC), and triglycerides (TGs)) and stone-forming factors (urinary pH, urinary calcium, and urinary oxalate acid) was carried out. In addition, the levels of oxidative stress (OS) markers (CAT, SOD, TAC, GSH-PX, and MDA) were measured, and histological analysis was carried out at the end of 16 weeks.
RESULTS
MS-related parameters, such as BW, LI, BG, TC, and TG, were significantly higher in HFD-fed rats than in RD-fed rats ( < 0.001). In the HFDD group, significantly lower urinary pH, hyperoxaluria, and hypocalciuria were noted in the dynamic assessment of stone-forming factors ( < 0.001). CAT, TAC, and MDA were notably changed in the HFD-fed groups, particularly the HFDD rats. Histological analysis showed that the renal tubules of HFDD rats had the highest scores for both inflammation and renal crystallization deposition ( < 0.05).
CONCLUSIONS
Our results suggest that male SD rats with MS are prone to developing nephrolithiasis. Validation in an model may lead to an understanding of the underlying pathophysiological mechanisms of action of MS-related nephrolithiasis in humans.Key messagesMale SD rats with metabolic syndrome are more prone to developing calcium oxalate nephrolithiasis after treatment with ethylene glycol and ammonium chloride compared to control lean rats.MS-related nephrolithiasis in rats induced by ethylene glycol and ammonium chloride is mainly related to increased hyperoxaluria and inflammation and decreased antioxidant levels.High-fat diet-fed SD rats treated with ethylene glycol and ammonium chloride are a stable and valid model for understanding the potential mechanism of action of MS-related nephrolithiasis.
Topics: Ammonium Chloride; Animals; Ethylene Glycol; Humans; Hyperoxaluria; Inflammation; Kidney Calculi; Male; Metabolic Syndrome; Nephrolithiasis; Rats; Rats, Sprague-Dawley
PubMed: 35837686
DOI: 10.1080/0886022X.2022.2097922 -
The Science of the Total Environment Sep 2023With the rapid development of industry and agriculture, excessive nitrogen and phosphorus released into natural surface water have caused eutrophication. Applying...
Effects of eutrophic water with ammonium chloride, urea, potassium dihydrogen phosphate and sodium-β-glycerophosphate on Myriophyllum verticillatum and epiphytic bacteria.
With the rapid development of industry and agriculture, excessive nitrogen and phosphorus released into natural surface water have caused eutrophication. Applying submerged plants to manage eutrophic water has attracted widespread attention. However, there are limited studies on the effects of different nitrogen and phosphorus in the water environment on submerged plants and their epiphytic biofilm. Therefore, this paper investigated the effects of eutrophic water with ammonium chloride (IN), urea (ON), potassium dihydrogen phosphate (IP), and sodium-β-glycerophosphate (OP) on Myriophyllum verticillatum and epiphytic biofilms. The results showed that Myriophyllum verticillatum exhibited a good purification effect on the eutrophic water with inorganic phosphorus, the removal rates of IP were 68.0%, and the plants grew best in this condition. The fresh weight of the IN group and ON group increased by 12.24% and 7.12%, and the shoot length of the IN group and the ON group increased by 17.71% and 8.33%; the fresh weight of the IP group and OP group increased by 19.19% and 10.83%, the shoot length of the IP group and the OP group increased by 21.09% and 18.23%. In addition, the enzyme activities of superoxide dismutase, catalase, nitrate reductase, and acid phosphatase in plant leaves were significantly changed in eutrophic water with different forms of nitrogen and phosphorus. Finally, the analysis of the epiphytic bacteria showed that different forms of nitrogen and phosphorus nutrients could significantly alter the abundance and structure of microorganisms and microbial metabolism also had significant changes. This study provides a new theoretical basis for evaluating the removal of different forms of nitrogen and phosphorus by Myriophyllum verticillatum, and it also provides new insights for the subsequent engineering of epiphytic microorganisms to improve the capability of submerged plants to treat eutrophic water.
Topics: Water; Ammonium Chloride; Urea; Bacteria; Saxifragales; Phosphorus; Plants; Nitrogen
PubMed: 37268138
DOI: 10.1016/j.scitotenv.2023.164507 -
Marine Drugs Sep 2018Hydroxypropyltrimethyl ammonium chitosan halogenated acetates were successfully synthesized from six different haloacetic acids and hydroxypropyltrimethyl ammonium...
Hydroxypropyltrimethyl ammonium chitosan halogenated acetates were successfully synthesized from six different haloacetic acids and hydroxypropyltrimethyl ammonium chloride chitosan (HACC) with high substitution degree, which are hydroxypropyltrimethyl ammonium chitosan bromacetate (HACBA), hydroxypropyltrimethyl ammonium chitosan chloroacetate (HACCA), hydroxypropyltrimethyl ammonium chitosan dichloroacetate (HACDCA), hydroxypropyltrimethyl ammonium chitosan trichloroacetate (HACTCA), hydroxypropyltrimethyl ammonium chitosan difluoroacetate (HACDFA), and hydroxypropyltrimethyl ammonium chitosan trifluoroacetate (HACTFA). These chitosan derivatives were synthesized by two steps: first, the hydroxypropyltrimethyl ammonium chloride chitosan was synthesized by chitosan and 3-chloro-2-hydroxypropyltrimethyl ammonium chloride. Then, hydroxypropyltrimethyl ammonium chitosan halogenated acetates were synthesized via ion exchange. The structures of chitosan derivatives were characterized by Fourier transform infrared spectroscopy (FTIR), ¹H Nuclear magnetic resonance spectrometer (¹H NMR), C Nuclear magnetic resonance spectrometer (C NMR), and elemental analysis. Their antifungal activities against , , , and were investigated by hypha measurement in vitro. The results revealed that hydroxypropyltrimethyl ammonium chitosan halogenated acetates had better antifungal activities than chitosan and HACC. In particular, the inhibitory activity decreased in the order: HACTFA > HACDFA > HACTCA > HACDCA > HACCA > HACBA > HACC > chitosan, which was consistent with the electron-withdrawing property of different halogenated acetates. This experiment provides a potential idea for the preparation of new antifungal drugs by chitosan.
Topics: Acetates; Antifungal Agents; Ascomycota; Carbon-13 Magnetic Resonance Spectroscopy; Chitosan; Fungicides, Industrial; Halogenation; Hyphae; Microbial Sensitivity Tests; Mitosporic Fungi; Molecular Structure; Plant Diseases; Proton Magnetic Resonance Spectroscopy; Quaternary Ammonium Compounds; Spectroscopy, Fourier Transform Infrared; Structure-Activity Relationship
PubMed: 30189609
DOI: 10.3390/md16090315 -
Clinical Microbiology and Infection :... Apr 2022To investigate the effect of daily whole-body bathing (WBB) using disposable washcloth wipes/caps impregnated with an antiseptic solution containing the quaternary... (Randomized Controlled Trial)
Randomized Controlled Trial
Effect of didecyl dimethyl ammonium chloride-impregnated washcloth wipe whole-body bathing on catheter-related blood stream infections and central venous line-associated infections in adult intensive care units.
OBJECTIVE
To investigate the effect of daily whole-body bathing (WBB) using disposable washcloth wipes/caps impregnated with an antiseptic solution containing the quaternary ammonium base compound didecyl dimethyl ammonium chloride (DDAC).
METHODS
A prospective double-blind randomized crossover trial was conducted to compare WBB of adult intensive care unit (ICU) patients with washcloth wipes/caps impregnated with either regular cleanser/shampoo or the antiseptic DDAC. The clinical trial was performed in a medical ICU (MICU) and a surgical ICU (SICU). The study period was divided into two 6-month intervals with alternating treatment regimens.
RESULTS
A total of 1540 ICU patients (total length of ICU stay 10 470 days) were included in the trial. Compared to controls, DDAC bathing was found to be associated with reduced incidence rates per 1000 patient days for central-venous-line-associated infections (CLAIs) caused by Gram-positive bacteria (GPB) from 16.39 (95%CI 13.1-20.3) in the control group to 7.28 (95%CI 5.2-9.9) in the intervention group (p 0.01). A stratified analysis by unit showed that the incidence rates of CLAI due to GPB were reduced by the intervention in both the MICU and the SICU from 21.2 (95%CI 15.8-27.7) to 9.3 (95%CI 5.8-14.1) (p < 0.01) and from 12.1 (95%CI 8.3-17.0) to 5.7 (95%CI 3.4-9.1) (p 0.01), respectively. There was a trend towards reduction in catheter-related bloodstream infections (CRBSIs) and bloodstream infections (BSIs); however, this did not reach statistical significance due to carry-over effects and small numbers.
CONCLUSIONS
Given the growing need for new concepts to prevent and control healthcare-associated infections, DDAC may be a new and promising agent for WBB of ICU patients.
Topics: Adult; Ammonium Chloride; Catheter-Related Infections; Catheterization, Central Venous; Catheters; Chlorhexidine; Chlorides; Cross Infection; Humans; Intensive Care Units; Prospective Studies
PubMed: 34333129
DOI: 10.1016/j.cmi.2021.07.029 -
European Journal of Pharmacology Sep 2019We performed a comparative analysis of molecular cytotoxic mechanisms of lysosomal autophagy inhibitors bafilomycin A1, chloroquine, and ammonium chloride in B16 mouse... (Comparative Study)
Comparative Study
We performed a comparative analysis of molecular cytotoxic mechanisms of lysosomal autophagy inhibitors bafilomycin A1, chloroquine, and ammonium chloride in B16 mouse melanoma cells. All agents caused oxidative stress, mitochondrial depolarization, and caspase-dependent apoptotic death, which was not affected by genetic inactivation of autophagy. Cathepsin inhibition reduced only the cytotoxicity of chloroquine, indicating its ability to cause lysosomal membrane permeabilization. Bafilomycin reduced the mRNA levels of anti-apoptotic Bcl-2, while chloroquine and ammonium chloride increased the mRNA expression of pro-apoptotic Pten and Puma, as well as anti-apoptotic Bcl-xL. Ammonium chloride additionally increased the mRNA expression of pro-apoptotic Bim and p53. All three agents decreased the activity of mechanistic target of rapamycin (mTOR) and increased the activation of p38 mitogen-activated protein kinase (MAPK). Chloroquine and ammonium chloride additionally stimulated the phosphorylation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), respectively, while only bafilomycin increased the phosphorylation of the energy sensor AMP-activated protein kinase (AMPK). mTOR activator leucine did not affect the cytotoxicity of lysosomal inhibitors. p38 MAPK inhibitor SB203580 reduced the cytotoxicity of bafilomycin but increased that of chloroquine and ammonium chloride. The pharmacological inhibition of ERK1/2, JNK, and AMPK potentiated the cytotoxicity of chloroquine, ammonium chloride, and bafilomycin, respectively. The observed mechanistic differences were associated with antagonistic interactions of lysosomal inhibitors in B16 cell killing. In conclusion, all investigated lysosomal inhibitors cause autophagy-independent mitochondrial dysfunction and apoptotic death, but differ in the ability to affect lysosomal permeabilization, balance between pro- and anti-apoptotic molecules of Bcl-2 family, and MAPK/AMPK signaling.
Topics: Animals; Antineoplastic Agents; Apoptosis; Autophagy; Cell Line, Tumor; Cell Survival; Lysosomes; MAP Kinase Signaling System; Melanoma, Experimental; Membrane Potential, Mitochondrial; Mice; Oxidative Stress
PubMed: 31310755
DOI: 10.1016/j.ejphar.2019.172540 -
Bulletin of Experimental Biology and... Feb 2020Ammonium, an end-product of catabolism, in low doses can promote adaptation of metabolic pathways in erythrocytes under conditions of extreme physical exercise. We...
Ammonium, an end-product of catabolism, in low doses can promote adaptation of metabolic pathways in erythrocytes under conditions of extreme physical exercise. We compared the effects of two ammonium salts, ammonium chloride and ammonium carbonate, in two doses on biochemical parameters of rat erythrocytes 1 day after extreme physical exercise in a 4-week cycle of forced swimming. Of 16 analyzed parameters, the maximum number of significant shifts from the control was revealed in the groups of rats receiving ammonium chloride in doses of 20 and 10 mg/kg, and the minimal number of differences was found in groups treated with ammonium carbonate in the same doses. The comparison of the levels of reduced glutathione and 2.3-bisphosphoglicerate and activities of 5'-nucleotidase and Ca- and Na/K-ATPases attested to more rigorous control of the mechanism of oxygen delivery to tissues by erythrocytes after administration of ammonium chloride in a dose of 20 mg/kg.
Topics: 2,3-Diphosphoglycerate; 5'-Nucleotidase; Adaptation, Physiological; Ammonium Chloride; Animals; Antioxidants; Calcium-Transporting ATPases; Carbonates; Dose-Response Relationship, Drug; Erythrocytes; Gene Expression; Glutathione; Oxidative Stress; Physical Conditioning, Animal; Physical Exertion; Rats; Sodium-Potassium-Exchanging ATPase; Swimming
PubMed: 32146621
DOI: 10.1007/s10517-020-04728-6