Sulfate Gradient Research Articles

Hybrid Nanosystem Formed by DOX-Loaded Liposomes and Extracellular Vesicles from MDA-MB-231 Is Effective against Breast Cancer Cells with Different Molecular Profiles.

Drug delivery selectivity is a challenge for cancer treatment. A hybrid pegylated pH-sensitive liposome-extracellular vesicle isolated from human breast cancer cell MDA-MB-231 was developed to investigate its in vitro activity against breast cancer cells of different molecular profiles to overcome this inconvenience. The hybrid nanosystem was produced by film hydration, and doxorubicin (DOX) was encapsulated in this system using the ammonium sulfate gradient method. The characterization of this hybrid nanosystem revealed a mean diameter of 140.20 ± 2.70 nm, a polydispersity index of 0.102 ± 0.033, an encapsulation efficiency of doxorubicin of 88.9% ± 2.4, and a great storage stability for 90 days at 4 °C. The fusion of extracellular vesicles with liposomes was confirmed by nanoflow cytometry using PE-conjugated human anti-CD63. This hybrid nanosystem demonstrated cytotoxicity against human breast cancer cell lines with different molecular subtypes, enhanced anti-migration properties, and exhibited similar cellular uptake to the free DOX treatment. Preliminary acute toxicity assessments using Balb/C female mice indicated a median lethal dose of 15-17.5 mg/kg, with no evidence of splenic, liver, heart, bone marrow, and renal damage at a dose of 15 mg/kg. These findings suggest the hybrid formulation as a versatile nanocarrier for the treatment of various breast cancer subtypes.

Application of targeted liposomes-based salvianolic acid A for the treatment of ischemic stroke

Novel therapeutics for the treatment of ischemic stroke remains to be the unmet clinical needs. Previous studies have indicated that salvianolic acid A (SAA) is a promising candidate for the treatment of the brain diseases. However, SAA has poor absolute bioavailability and does not efficiently cross the intact blood-brain barrier (BBB), which limit its efficacy. To this end we developed a brain-targeted liposomes for transporting SAA via the BBB by incorporating the liposomes to a transport receptor, insulin-like growth factor-1 receptor (IGF1R). The liposomes were prepared by ammonium sulfate gradients loading method. The prepared SAA-loaded liposomes (Lipo/SAA) were modified with IGF1R monoclonal antibody to generate IGF1R antibody-conjugated Lipo/SAA (IGF1R-targeted Lipo/SAA). The penetration of IGF1R-targeted Lipo/SAA into the brain was confirmed by labeling with Texas Red, and their efficacy were evaluate using middle cerebral artery occlusion (MCAO) model. The results showed that IGF1R-targeted Lipo/SAA are capable of transporting SAA across the BBB into the brain, accumulation in brain tissue, and sustained releasing SAA for several hours. Administration o IGF1R-targeted Lipo/SAA notably reduced infarct size and neuronal damage, improved neurological function and inhibited cerebral inflammation, which had much higher efficiency than no-targeted SAA.

Electrochemical reconstruction of non-noble metal-based heterostructure nanorod arrays electrodes for highly stable anion exchange membrane seawater electrolysis

Direct seawater electrolysis for hydrogen production has been regarded as a viable route to utilize surplus renewable energy and address the climate crisis. However, the harsh electrochemical environment of seawater, particularly the presence of aggressive Cl−, has been proven to be prone to parasitic chloride ion oxidation and corrosion reactions, thus restricting seawater electrolyzer lifetime. Herein, hierarchical structure (Ni,Fe)O(OH)@NiCoS nanorod arrays (NAs) catalysts with heterointerfaces and localized oxygen vacancies were synthesized at nickel foam substrates via the combination of hydrothermal and annealing methods to boost seawater dissociation. The hierarchical nanostructure of NiCoS NAs enhanced electrode charge transfer rate and active surface area to accelerate oxygen evolution reaction (OER) and generated sulfate gradient layers to repulsive aggressive Cl−. The fabricated heterostructure and vacancies of (Ni,Fe)O(OH) tuned catalyst electronic structure into an electrophilic state to enhance the binding affinity of hydroxyl intermediates and facilitate the structural transformation into amorphous γ-NiFeOOH for promoting OER. Furthermore, through operando electrochemistry techniques, we found that the γ-NiFeOOH possessing an unsaturated coordination environment and lattice-oxygen-participated OER mechanism can minimize electrode Cl− corrosion enabled by stabilizing the adsorption of OH* intermediates, making it one of the best OER catalysts in the seawater medium reported to date. Consequently, these catalysts can deliver current densities of 100 and 500 mA cm−2 for boosting OER at minimal overpotentials of 245 and 316 mV, respectively, and thus prevent chloride ion oxidation simultaneously. Impressively, a highly stable anion exchange membrane (AEM) seawater electrolyzer based on the non-noble metal heterostructure electrodes reached a record low degradation rate under 100 μV h−1 at constant industrial current densities of 400 and 600 mA cm−2 over 300 h, which exhibits a promising future for the non-precious and stable AEMWE in the direct seawater electrolysis industry.

Remote Loading: The Missing Piece for Achieving High Drug Payload and Rapid Release in Polymeric Microbubbles.

The use of drug-loaded microbubbles for targeted drug delivery, particularly in cancer treatment, has been extensively studied in recent years. However, the loading capacity of microbubbles has been limited due to their surface area. Typically, drug molecules are loaded on or within the shell, or drug-loaded nanoparticles are coated on the surfaces of microbubbles. To address this significant limitation, we have introduced a novel approach. For the first time, we employed a transmembrane ammonium sulfate and pH gradient to load doxorubicin in a crystallized form in the core of polymeric microcapsules. Subsequently, we created remotely loaded microbubbles (RLMBs) through the sublimation of the liquid core of the microcapsules. Remotely loaded microcapsules exhibited an 18-fold increase in drug payload compared with physically loaded microcapsules. Furthermore, we investigated the drug release of RLMBs when exposed to an ultrasound field. After 120 s, an impressive 82.4 ± 5.5% of the loaded doxorubicin was released, demonstrating the remarkable capability of remotely loaded microbubbles for on-demand drug release. This study is the first to report such microbubbles that enable rapid drug release from the core. This innovative technique holds great promise in enhancing drug loading capacity and advancing targeted drug delivery.

Preparation of liposomes co-encapsulating doxorubicin and mifepristone for treating multidrug-resistant cancer

The emergence of multidrug resistance (MDR) is well-known to decrease the therapeutic effects of anticancer drugs. To increase anticancer drug efficacy and decrease adverse effects, we prepared liposomes co-encapsulating doxorubicin (DXR) and mifepristone (MIF) (DXR/MIF-LPs) for treating MDR cancer and evaluated the MDR reversal effect in DXR-resistant Colon-26 cancer cells (C26/DXR). Liposomal encapsulation was performed using remote loading methods: amphipathic DXR was encapsulated using the ammonium sulfate gradient method, whereas hydrophobic MIF was encapsulated in preformed liposomes by incubating with water-miscible dimethyl sulfoxide. Both drugs were successfully encapsulated in liposomes by simultaneously incubating with blank liposomes without remarkably altering the particle size. The release of each drug from DXR/MIF-LPs corresponded to that from liposomes encapsulating a single drug, indicating that drug encapsulation did not influence drug release. Treatment with MIF did not impact the overexpression of P-glycoprotein (P-gp) in C26/DXR cells. However, DXR/MIF-LPs increased the cellular association of DXR and enhanced cytotoxicity compared with liposomes encapsulating DXR against C26/DXR cells, indicating that MIF inhibits P-gp function and increases the cytotoxicity of DXR in MDR cells. Collectively, our findings suggest that the co-delivery of DXR and MIF using a liposomal formulation is a promising approach for reversing MDR.

Functionalized curcumin/ginsenoside Rb1 dual-loaded liposomes: Targeting the blood-brain barrier and improving pathological features associated in APP/PS-1 mice

The brain is a crucial organ in the body, shielded from external substances by the blood-brain barrier (BBB). However, this barrier also impedes the treatment of numerous brain diseases, including Alzheimer's disease (AD). Therefore, Mannose (MAN)-modified co-loaded Curcumin (CUR) and ginsenoside Rb1 (Rb1) liposomes were prepared to address the above situation. Glucose transporter 1 (GLUT 1) is a transmembrane glycoprotein overexpressed on the BBB. MAN is specifically recognized and efficiently transported by GULT1. In addition, the neuroprotective compounds CUR and Rb1 are co-loaded into liposomes, which can penetrate the blood-brain barrier (BBB) and potentially be used to treat AD. MAN-CUR/Rb1 liposomes were prepared using a thin film hydration method combined with an ammonium sulfate gradient. The resulting liposomes had a round shape and a diameter of 110 nm and showed good stability. Results from cellular uptake experiments, in vitro BBB models, and in vivo imaging in mice confirmed that MAN-CUR/Rb1 liposomes not only crossed the BBB but also accumulate in N2a cells/bEnd.3 cells. Furthermore, in vivo animal experiments showed that MAN-CUR/Rb1 liposomes improved AD-related pathological features, inhibited oxidative stress and neuroinflammation, and enhanced learning and cognition in APP/PS-1 mice. Based on these findings, MAN-CUR/Rb1 liposomes hold promise as a potential treatment for AD.

Controlling the Evolution of Selective Vancomycin Resistance through Successful Ophthalmic Eye-Drop Preparation of Vancomycin-Loaded Nanoliposomes Using the Active-Loading Method.

Vancomycin is the front-line defense and drug of choice for the most serious and life-threatening methicillin-resistant Staphylococcus aureus (MRSA) infections. However, poor vancomycin therapeutic practice limits its use, and there is a consequent rise of the threat of vancomycin resistance by complete loss of its antibacterial activity. Nanovesicles as a drug-delivery platform, with their featured capabilities of targeted delivery and cell penetration, are a promising strategy to resolve the shortcomings of vancomycin therapy. However, vancomycin's physicochemical properties challenge its effective loading. In this study, we used the ammonium sulfate gradient method to enhance vancomycin loading into liposomes. Depending on the pH difference between the extraliposomal vancomycin-Tris buffer solution (pH 9) and the intraliposomal ammonium sulfate solution (pH 5-6), vancomycin was actively and successfully loaded into liposomes (up to 65% entrapment efficiency), while the liposomal size was maintained at 155 nm. Vancomycin-loaded nanoliposomes effectively enhanced the bactericidal effect of vancomycin; the minimum inhibitory concentration (MIC) value for MRSA decreased 4.6-fold. Furthermore, they effectively inhibited and killed heteroresistant vancomycin-intermediate S.aureous (h-VISA) with an MIC of 0.338 μg mL-1. Moreover, MRSA could not develop resistance against vancomycin that was loaded into and delivered by liposomes. Vancomycin-loaded nanoliposomes could be a feasible solution for enhancing vancomycin's therapeutic use and controlling the emerging vancomycin resistance.

Anticancer Effect of STING Agonist-Encapsulated Liposomes on Breast Cancer.

Breast cancer is one of the most common cancers worldwide, posing a serious threat to human health. Recently, innate immunity has become a widely discussed topic in antitumor research. The STING pathway is an important component of innate immunity, and several STING agonists have been developed and applied in antitumor research. Dimeric amidobenzimidazole (diABZI) is one STING agonist and is a nucleotide analog with low serological stability and cell membrane permeability. In this study, we prepared diABZI-encapsulated liposomes (dLNPs) using the ammonium sulfate gradient method. The average particle size of the dLNPs was 99.76 ± 0.230 nm, and the encapsulation efficiency was 58.29 ± 0.53%. Additionally, in vivo and in vitro assays showed that the dLNPs had a sustained-release effect and that the circulation time in vivo was longer than 48 h. The expression of IFN-β and IFN-γ was elevated in mice treated with dLNPs. Moreover, we found that dLNPs can recruit CD8+ T cells to tumor tissue and exert antitumor effects. The dLNPs-treated group showed the most significant efficacy: the average tumor volume was 231.46 mm3, which decreased by 78.16% and 54.47% compared to the PBS group and diABZI group. Meanwhile, the hemolysis rate of the dLNPs was 2%, showing high biocompatibility. In conclusion, dLNPs can effectively suppress tumor growth and possess great potential in breast cancer therapy.

Preparation and intraocular evaluation of cetirizine hydrochloride ophthalmic liposomes and a liposome in situ gel

AbstractCetirizine hydrochloride (CTZ), an antiallergic drug, is a new‐generation H1 receptor antagonist and a second‐generation H1 antihistamine. We aimed to prepare cetirizine hydrochloride liposomes, based on which cetirizine hydrochloride liposomal (CTZL) in situ gel (ISG) was prepared, to improve the retention time in the eye. CTZL were prepared by the ethanol injection method combined with the ammonium sulfate gradient method. A CTZ liposomal temperature‐sensitive gel was prepared using the cold dissolution method. Large‐eared white rabbits were used in retention and irritation experiments. The liposomes were small single‐chambered liposomes, spherical or sphere‐like, with a vesicle size of 187.03 ± 6.20 nm, an encapsulation efficiency of 70.39 ± 1.13%, and a drug loading capacity of 4.63 ± 0.06%. The gelling temperatures before and after dilution by simulated tear fluid were 26.1 ± 0.2°C and 34.2 ± 0.2°C, the vesicle size was 184.94 ± 7.28 nm, and the liposomes were spherical or sphere‐like in the gel matrix. The in vitro dissolution and release experiments indicate that the gel was released upon dissolution and exhibited a zero‐level release pattern. Preparation into liposomes and liposomal gels prolonged the ocular retention time of the formulation without ocular irritation.

Environmental formation of methylmercury is controlled by synergy of inorganic mercury bioavailability and microbial mercury-methylation capacity.

Methylmercury (MeHg) production is controlled by the bioavailability of inorganic divalent mercury (Hg(II)i ) and Hg-methylation capacity of the microbial community (conferred by the hgcAB gene cluster). However, the relative importance of these factors and their interaction in the environment remain poorly understood. Here, metagenomic sequencing and a full-factorial MeHg formation experiment were conducted across a wetland sulfate gradient with different microbial communities and pore water chemistries. From this experiment, the relative importance of each factor on MeHg formation was isolated. Hg(II)i bioavailability correlated with the dissolved organic matter composition, while the microbial Hg-methylation capacity correlated with the abundance of hgcA genes. MeHg formation responded synergistically to both factors. Notably, hgcA sequences were from diverse taxonomic groups, none of which contained genes for dissimilatory sulfate reduction. This work expands our understanding of the geochemical and microbial constraints on MeHg formation in situ and provides an experimental framework for further mechanistic studies. This article is protected by copyright. All rights reserved.

Preparation of an HI-6-loaded brain-targeted liposomes based on the nasal delivery route and the evaluation of its reactivation of central toxic acetylcholinesterase

PurposeOrganophosphorus compounds (OPs) is a serious threat to human health and life safety, but because of the existence of blood-brain barrier, most of the therapeutic drugs cannot enter the center, reactivate centrally located toxic acetylcholinesterase (AChE), it is urgent to find an efficient treatment method. MethodsThe c(RGDyK) cyclic peptide modified HI-6-loaded brain targeting liposomes [c(RGDyK)-PEG2000HI-6-lipo] were prepared by ammonium sulfate gradient method. The in vitro blood-brain barrier (BBB) model was established, and the function of the liposomes was evaluated. The animal model of DDVP poisoning was established, and the central toxic enzyme reactivation ability of c(RGDyK)-PEG2000HI-6-lipo by both the intravenous and nasal administration route was verified. ResultsThe HI-6-loaded liposomes with brain targeting function were successfully synthesized and prepared with high encapsulation efficiency (70.23 ± 2.18%), drug loading (2.86 ± 0.07)%, average particle size 242.9 nm (polydispersion index 0.149), and ζ potential -16.2 mV. Combined with the in vitro and in vivo studies, the c(RGDyK)-PEG2000HI-6-lipo has better ability to cross the BBB. In addition, compared with intravenous injection, nasal administration was proved to be more effective against organophosphorus poisoning, and the reactivation rate of brain acetylcholinesterase reached (26.19 ± 7.70)%. ConclusionThe prepared c(RGDyK)-PEG2000HI-6-lipo has a better ability to cross BBB. Nasal administration, as a way to bypass the BBB and directly deliver drugs into the brain, effectively improves the bioavailability of HI-6 in the brain. This study holds promise by providing a non-invasive approach to deliver water-soluble oxime antidote into the brain and reactivate central acetylcholinesterase via the naso-brain route.

Surface-decorated nanoliposomal leonurine targets activated fibroblast-like synoviocytes for efficient rheumatoid arthritis therapy.

Rheumatoid arthritis (RA) is a chronic autoimmune disease that causes progressive joint destruction, leading to impaired life quality, disability, and even premature mortality. However, current medications suffer from limited clinical outcomes and severe side effects due to low bioavailability and non-specific distribution after administration. Herein, a targeting nanosystem (HAP-Lipo@Leo) was constructed for efficient RA treatment, which can precisely deliver a natural anti-arthritic drug leonurine (Leo) to the inflamed joint by HAP-1 peptide-mediated recognition of activated fibroblast-like synoviocytes (FLS). More specifically, HAP-Lipo@Leo was prepared by a combination of thin film hydration and high-pressure microfluidization and surface-decorated with HAP-1 peptide and PEG before encapsulating Leo by the ammonium sulfate gradient method. The as-obtained HAP-Lipo@Leo can be selectively internalized by activated FLS and impairs the lamellipodia formation and overexpression of inflammatory cytokines, both of which play detrimental roles in joint damage. Furthermore, HAP-Lipo@Leo demonstrated arthritic joint-specific distribution, significant inhibition of synovial inflammation, and reversal of cartilage and bone destruction in adjuvant-induced arthritis rats as evidenced by comprehensive investigations including ELISA tests, histopathology examinations, and micro-CT analysis. In addition, HAP-Lipo@Leo exhibited good biocompatibility and safety both in vitro and in vivo. Taken together, HAP-Lipo@Leo holds great potential for clinical RA management by integrating activated FLS targeting, long circulation, multifaceted therapeutic effects, and excellent biocompatibility.

A protocol for setting-up robust hydrophobic interaction chromatography targeting the analysis of intact proteins and monoclonal antibodies.

Hydrophobic interaction chromatography (HIC) is a chromatographic technique that mainly targets the separation of biomolecules (intact proteins, monoclonal antibodies, etc.) based on the difference in surface hydrophobicity while applying non-denaturing conditions. This protocol paper provides guidelines for setting-up robust HIC analysis and considers the instrument configuration, mobile-phase and sample preparation, as well as chromatographic conditions and settings. The separation of a mixture of intact proteins and monoclonal antibodies is demonstrated by applying conventional HIC conditions, that is, using a mildly hydrophobic (C4) stationary phase in combination with an inverse ammonium sulphate gradient dissolved in aqueous phosphate buffer. The effect of sample-preparation conditions on sample breakthroughs is presented. Finally, good run-to-run repeatability (relative standard deviation<2%) is demonstrated for five different columns obtained from three different column lots, considering chromatographic retention, peak width, peak area and column pressure.

Doxorubicin-liposome combined with clodronate-liposome inhibits hepatocellular carcinoma through the depletion of macrophages and tumor cells

Macrophages in the liver have capacities of capturing and phagocytosing nanocarriers. Macrophages also play an important role in the inflammatory microenvironment and in the tumorigenesis, development and progression of hepatocellular carcinoma (HCC). Several studies have shown that depletion of macrophages is a viable strategy for drug delivery and tumor microenvironment regulation. We prepared liposomes containing doxorubicin and clodronate using an ammonium sulfate gradient and thin film hydration method. The repressive therapeutic effects of liposomes were compared by intrasplenic injection at different stages of a primary HCC model induced by diethylnitrosamine (DEN) in rats. Doxorubicin-liposome (DOX-LIP) and clodronate-liposome (CL-LIP) about 180–200 nm were successfully prepared and characterized. We found that DOX-LIP combined with CL-LIP could effectively inhibit the occurrence and development of liver cancer without major organ damage and side effects. The combination of doxorubicin and clodronate liposomes notably decreased hepatic CD68 + macrophages, enriched DOX in plasma and accumulated it for a long time in the liver and spleen, thus improving the tumor microenvironment, inhibiting the activation of hepatic progenitor cells (HPCs) and promoting the apoptosis of tumor cells, and finally producing the inhibitory and therapeutic effects of HCC in rats. Results of this study were expected to provide a new prospect for the chemotherapy of HCC.

Exosome mimetics derived from bone marrow mesenchymal stem cells ablate neuroblastoma tumor in vitro and in vivo

PurposeTo develop exosome-mimetics derived from bone marrow mesenchymal stem cells (EM) as a novel nanoscale drug delivery system(nanoDDS) with improved tumor targeting activity, therapeutic effect, and biosafety, and to evaluate the therapeutic effect of doxorubicin loaded EM (EM-Dox) on neuroblastoma (NB) in vitro and in vivo. MethodsEM was prepared by serial extrusion of bone marrow mesenchymal stem cells (BMSCs), ammonium sulfate gradient method was used to promote the active loading of doxorubicin, and EM-Dox was obtained after removal of free doxorubicin by dialysis. The obtained EM and EM-Dox were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), Western Blot assay(WB), and the yield of exosomes and EM was further compared. Confocal fluorescent microscopy was used to verify the uptake of EM-Dox and free doxorubicin (Free-Dox) by NB cells. CCK-8 assay, cell cycle assay, and cell apoptosis assay were used to evaluate the antitumor effect of EM-Dox on NB cells in vitro. In addition, the targeted therapeutic effect and biosafety of EM-Dox against NB were evaluated in tumor-bearing nude mice. ResultsTEM, NTA, and WB verified that both EM and EM-Dox feature highly similar morphology, size and marker protein expression in comparison with naturally occurred exosomes, but the particle size of EM-Dox increased slightly after loading doxorubicin. The protein yield and particle yield of EM-Dox were 16.8 and 26.3-folds higher than those of exosomes, respectively. Confocal fluorescent microscopy showed that EM and doxorubicin had a definite co-localization. EM-Dox was readily internalized in two well-established human NB cell lines. The intracellular content of doxorubicin in cells treated with EM-Dox was significantly higher than that treated with Free-Dox. CCK-8 assay and flow cytometry confirmed that EM-Dox could inhibit NB cell proliferation, induce G2/M phase cell cycle arrest, and promote NB cell apoptosis in vitro. In vivo bioluminescence imaging results demonstrated that EM-Dox effectively targets NB tumors in vivo. Compared with Free-Dox, EM-Dox had a significantly increased inhibitory effect against NB tumor proliferation and progression in vivo, without inducing any myocardial injury. ConclusionsEM-Dox showed significantly increased anti-tumor activity in comparison with free doxorubicin in vitro and in vivo, and scalable EMs may represent a new class of NanoDDS that can potentially replace naturally occurred exosomes in preclinical or clinical translations.

Integrating water quality monitoring and diatom community trends to determine landscape‐level change in protected lakes

AbstractLakes in protected or remote regions are not immune to anthropogenic impacts and face a variety of stressors ranging from atmospheric deposition of pollutants to global climate change. Monitoring programs in these lakes can be limited in scope due to sampling logistics, leaving an incomplete picture of how the systems may be impacted by environmental stressors. Here, we used diatoms as early indicators of change in relatively undisturbed lakes from five national park units in the US Great Lakes region. Surface sediment samples were collected repeatedly over more than a decade to analyze the diatom community turnover. This diatom community assemblage data were compared with the measured water quality data collected over the same period in order to identify the predominant drivers of ecological change. Even though the parks in this study span two biomes, and lake characteristics vary between and within parks, we found synchronicity in measured water quality trends and diatom responses. Changes in the thermal regime and water‐column mixing appeared to drive much of the change across parks, although much of the diatom turnover also followed a sulfate or pH gradient. Nutrients did not appear to play a major role in diatom community change. This method of using diatoms in conjunction with water quality monitoring allows for an integrated response over a number of years and provides managers with a complementary tool to determine which environmental parameters are having the biggest effect on lake ecology.

Pore-Water Geochemical Gradients of Sulfate, Calcium, Magnesium, and Iodide Correlated With Underlying Gas Hydrate Potential: A Case Study of the Shenhu Area, South China Sea

Geochemical profiles in pore water of marine sediments have been considered as, important indicators of gas hydrate occurrence. In the gas hydrate area around the world, the decrease of sulfate, calcium, and magnesium concentrations with depth mainly results directly or indirectly from the anaerobic oxidation of methane (AOM). The ubiquitous abnormally high concentration gradients of iodide in the research area reflect the large methane-generating potential of the area. Thus, we explore the feasibility of using gradients of sulfate, iodide, and authigenic carbonate precipitation as indicators for gas hydrate in the regional exploration of gas hydrate formation. We test the criterion in the gas hydrate zone in the South China Sea (SCS) for the recognition of gas hydrate by using the gradients of sulfate, calcium plus magnesium, and iodide. Contour maps of pore-water gradients from expeditions in the study area are used to correlate the key gradients to underlying gas hydrate occurrence. The results show that the largest potential gas hydrate indicated by the contour maps of the indicators are well consistent with the discovery of GMGS1 and GMGS3 expedition. It implies the possible applicability of this geochemical method in gas hydrate exploration. Also, we identify a promising area in the South China Sea for future gas hydrate investigations. It is the first collective application of the gradients of sulfate, calcium plus magnesium, and iodide to a gas hydrate terrane, especially to a large area of the SCS. We believe that the result of this research will benefit the future exploration of gas hydrate and will arouse a lot of interest from other researchers.

Chitosan-Coated Liposomes: The Strategy to Reduce Intestinal Toxicity and Improve Bioavailability of Oral Vinorelbine.

In recent years, the oral administration of vinorelbine has gradually replaced intravenous administration in the treatment of several types of tumors. Even though the risk of phlebitis is avoided with oral administration, oral vinorelbine is still not a highly patient-compliant route due to the severe gastrointestinal toxicity. Vinorelbine-loaded liposomes with high encapsulation efficiency and suitable particle size were prepared using the ammonium sulfate gradient method. Chitosan-coated liposomes showed the slowest in vitro release compared to uncoated liposomes and vinorelbine solution. No damage was observed in the intestinal epithelial cells of mice orally administered with coated vinorelbine liposomes due to the low presence of the free drug in the gastrointestinal tract and the LD50 was increased from 129.83 to 182.25mg/kg compared to oral vinorelbine solution. In addition, the positive surface potential of chitosan-coating endowed liposomes with mucosal adhesive function, delaying the time to reach the peak plasma concentration of vinorelbine from 1 to 4h after administration. And bioavailability was increased to 2.1-fold compared to vinorelbine solution. In short, a new strategy to address the severe gastrointestinal side effects of oral vinorelbine has been developed.

Active anaerobic methane oxidation and sulfur disproportionation in the deep terrestrial subsurface

Microbial life is widespread in the terrestrial subsurface and present down to several kilometers depth, but the energy sources that fuel metabolism in deep oligotrophic and anoxic environments remain unclear. In the deep crystalline bedrock of the Fennoscandian Shield at Olkiluoto, Finland, opposing gradients of abiotic methane and ancient seawater-derived sulfate create a terrestrial sulfate-methane transition zone (SMTZ). We used chemical and isotopic data coupled to genome-resolved metaproteogenomics to demonstrate active life and, for the first time, provide direct evidence of active anaerobic oxidation of methane (AOM) in a deep terrestrial bedrock. Proteins from Methanoperedens (formerly ANME-2d) are readily identifiable despite the low abundance (≤1%) of this genus and confirm the occurrence of AOM. This finding is supported by 13C-depleted dissolved inorganic carbon. Proteins from Desulfocapsaceae and Desulfurivibrionaceae, in addition to 34S-enriched sulfate, suggest that these organisms use inorganic sulfur compounds as both electron donor and acceptor. Zerovalent sulfur in the groundwater may derive from abiotic rock interactions, or from a non-obligate syntrophy with Methanoperedens, potentially linking methane and sulfur cycles in Olkiluoto groundwater. Finally, putative episymbionts from the candidate phyla radiation (CPR) and DPANN archaea represented a significant diversity in the groundwater (26/84 genomes) with roles in sulfur and carbon cycling. Our results highlight AOM and sulfur disproportionation as active metabolisms and show that methane and sulfur fuel microbial activity in the deep terrestrial subsurface.

Near-infrared light (NIR)-responsive nanoliposomes combining photodynamic therapy and chemotherapy for breast tumor control

Light-responsive carriers have been used for the controlled release of antitumor drugs in recent years. However, most light-responsive vectors require high-energy ultraviolet or visible light to achieve local drug release, and ultraviolet light would cause cellular damage. Near-infrared light has a deeper tissue-penetration depths and minimal harm to tissues, but it is difficult to cleave the chemical bond directly. The aim of this study is to develop a novel near-infrared light-responsive carrier for local release of antitumor drugs. Unsaturated phospholipids can be oxidized by singlet oxygen to achieve liposomal drug release, and singlet oxygen can be produced by photosensitizer under light irradiation. A new near-infrared light-responsive nanoliposome was designed that imparts light-triggered local drug release. Nanoliposomes, which were composed of matrix phospholipids and unsaturated phospholipids, were prepared by ammonium sulfate gradient method, and loaded with antitumor drug doxorubicin (DOX) and photosensitizer 1,4,8,11,15,18,22,25-octabutoxypalladium phthalocyanine. Under near-infrared light, photosensitizers could produce singlet oxygen and damage tumor cells by photodynamic therapy. Simultaneously, the unsaturated phospholipids were oxidized by singlet oxygen and result in DOX release, causing sustained cell damage by chemotherapy. Near-infrared light-responsive nanoliposomes exhibit enhanced anticancer activity owing to combined treatment of photodynamic therapy and chemotherapy. A new platform is thus offered for designing effective intracellular drug-release systems, holding great promise for future cancer therapy.