The Inhibition Effect of Linezolid With Reyanning Mixture on MRSA and its Biofilm is More Significant than That of Linezolid Alone

Methicillin-resistant Staphylococcus aureus (MRSA) is a superbacterium, and when it forms biofilms, it is difficult to treat even with the first-line of antibiotic linezolid (LNZ). Reyanning mixture (RYN), a compound-based Chinese medicine formula, has been found to have inhibitory effects on biofilms. This study aims to explore the synergistic inhibitory effect and corresponding mechanisms of their (LNZ&RYN) combination on the planktonic as well as biofilm cells of MRSA. Broth microdilution and chessboard methods were employed for the determination of minimum inhibitory concentrations (MICs) and synergistic concentration of LNZ&RYN, respectively. The effect of the combined medication on biofilm and mature biofilm of MRSA were observed by biofilm morphology and permeability experiments, respectively. To unveil the molecular mechanism of action of the synergistic combination of LNZ and RYN, RT-PCR based biofilm-related gene expression analysis and ultra-high pressure liquid chromatography-time-of-flight mass spectrometry based endogenous metabonomic analysis were deployed. The results indicated that 1/16RYN as the best combined dose reduced LNZ (4 μg/ml) to 2 μg/ml. The combined treatment inhibited living MRSA before and after biofilm formation, removed the residual structure of dead bacteria in MRSA biofilms and affected the shape and size of bacteria, resulting in the improvement of biofilm permeability. The mechanism was that biofilm-related genes such as agrC, atlA, and sarA, as well as amino acid uptake associated with the metabolism of 3-dehydrocarnitine, kynurenine, L-leucine, L-lysine and sebacic acid were inhibited. This study provides evidence for the treatment of MRSA and its biofilms with LNZ combined with RYN.

Modulation of Insulin Resistance, Dyslipidemia and Serum Metabolome in iNOS Knockout Mice following Treatment with Nitrite, Metformin, Pioglitazone, and a Combination of Ampicillin and Neomycin

Oxidative and nitrosative stress plays a pivotal role in the incidence of metabolic disorders. Studies from this lab and others in iNOS-/- mice have demonstrated occurrence of insulin resistance (IR), hyperglycemia and dyslipidemia highlighting the importance of optimal redox balance. The present study evaluates role of nitrite, L-arginine, antidiabetics (metformin, pioglitazone) and antibiotics (ampicillin-neomycin combination, metronidazole) on metabolic perturbations observed in iNOS-/- mice. The animals were monitored for glucose tolerance (IPGTT), IR (insulin, HOMA-IR, QUICKI), circulating lipids and serum metabolomics (LC-MS). Hyperglycemia, hyperinsulinemia and IR were rescued by nitrite, antidiabetics, and antibiotics treatments in iNOS-/- mice. Glucose intolerance was improved with nitrite, metformin and pioglitazone treatment, while ampicillin-neomycin combination normalised the glucose utilization in iNOS-/- mice. Increased serum phosphatidylethanolamine lipids in iNOS-/- mice were reversed by metformin, pioglitazone and ampicillin-neomycin; dyslipidemia was however marginally improved by nitrite treatment. The metabolic improvements were associated with changes in selected serum metabolites-purines, ceramide, 10-hydroxydecanoate, glucosaminate, diosmetin, sebacic acid, 3-nitrotyrosine and cysteamine. Bacterial metabolites-hippurate, indole-3-ethanol; IR marker-aminoadipate and oxidative stress marker-ophthalmate were reduced by pioglitazone and ampicillin-neomycin, but not by nitrite and metformin treatment. Results obtained in the present study suggest a crucial role of gut microbiota in the metabolic perturbations observed in iNOS-/- mice.

Clinical Significance of Screening Differential Metabolites in Ovarian Cancer Tissue and Ascites by LC/MS

Tumor cells not only show a vigorous metabolic state, but also reflect the disease progression and prognosis from their metabolites. To judge the progress and prognosis of ovarian cancer is generally based on the formation of ascites, or whether there is ascites recurrence during chemotherapy after ovarian cancer surgery. To explore the relationship between the production of ascites and ovarian cancer tissue, metabolomics was used to screen differential metabolites in this study. The significant markers leading to ascites formation and chemoresistance were screened by analyzing their correlation with the formation of ascites in ovarian cancer and the clinical indicators of patients, and then provided a theoretical basis. The results revealed that nine differential metabolites were screened out from 37 ovarian cancer tissues and their ascites, among which seven differential metabolites were screened from 22 self-paired samples. Sebacic acid and 20-COOH-leukotriene E4 were negatively correlated with the high expression of serum CA125. Carnosine was positively correlated with the high expression of serum uric acid. Hexadecanoic acid was negatively correlated with the high expression of serum γ-GGT and HBDH. 20a,22b-Dihydroxycholesterol was positively correlated with serum alkaline phosphatase and γ-GGT. In the chemotherapy-sensitive and chemotherapy-resistant ovarian cancer tissues, the differential metabolite dihydrothymine was significantly reduced in the chemotherapy-resistant group. In the ascites supernatant of the drug-resistant group, the differential metabolites, 1,25-dihydroxyvitamins D3-26, 23-lactonel and hexadecanoic acid were also significantly reduced. The results indicated that the nine differential metabolites could reflect the prognosis and the extent of liver and kidney damage in patients with ovarian cancer. Three differential metabolites with low expression in the drug-resistant group were proposed as new markers of chemotherapy efficacy in ovarian cancer patients with ascites.

Andrographolide Sulfonate Is a Promising Treatment to Combat Methicillin-resistant Staphylococcus aureus and Its Biofilms

Methicillin-resistant Staphylococcus aureus (MRSA) is a drug-resistant pathogen threatening human health and safety. Biofilms are an important cause of its drug resistance and pathogenicity. Inhibition and elimination of biofilms is an important strategy for the treatment of MRSA infection. Andrographolide sulfonate (AS) is an active component of the traditional herbal medicine Andrographis paniculata. This study aims to explore the inhibitory effect and corresponding mechanisms of AS on MRSA and its biofilms. Three doses of AS (6.25, 12.5, and 25 mg/ml) were introduced to MRSA with biofilms. In vitro antibacterial testing and morphological observation were used to confirm the inhibitory effect of AS on MRSA with biofilms. Real-time PCR and metabonomics were used to explore the underlying mechanisms of the effect by studying the expression of biofilm-related genes and endogenous metabolites. AS displayed significant anti-MRSA activity, and its minimum inhibitory concentration was 50 μg/ml. Also, AS inhibited biofilms and improved biofilm permeability. The mechanisms are mediated by the inhibition of the expression of genes, such as quorum sensing system regulatory genes (agrD and sarA), microbial surface components–recognizing adhesion matrix genes (clfA and fnbB), intercellular adhesion genes (icaA, icaD, and PIA), and a gene related to cellular eDNA release (cidA), and the downregulation of five biofilm-related metabolites, including anthranilic acid, D-lactic acid, kynurenine, L-homocitrulline, and sebacic acid. This study provided valuable evidence for the activity of AS against MRSA and its biofilms and extended the methods to combat MRSA infection.

Vancomycin-Induced Modulation of Gram-Positive Gut Bacteria And Metabolites Remediate Insulin Resistance In INOS Knockout Mice

Background: Inducible nitric oxide synthase (iNOS) has emerged as a crucial regulator of host metabolism and gut microbiota activity. The present study examines the role of the gut microbiome in determining host metabolic functions in absence of iNOS.Results: Insulin resistant and dyslipidemic iNOS-/- mice displayed reduced microbial diversity, with a higher relative abundance of the gram-positive bacteria, Allobaculum and Bifidobacterium, and altered serum metabolites linked with the metabolic dysregulation. Vancomycin, which largely depletes gram-positive bacteria, reversed the insulin resistance (IR), dyslipidemia, and related metabolic abnormalities in iNOS-/- mice. Such correction in metabolic markers was accompanied by reduced the expression of genes involved in fatty acid synthesis in liver and adipose tissue, decreased lipid uptake by adipose tissue and enhanced lipid efflux by liver and intestine. Rescue of IR in vancomycin treated iNOS-/- mice was associated with the alterations in the select serum metabolites such as 10-hydroxydecanoate, indole-3-ethanol, allantoin, hippurate, sebacic acid, aminoadipate, and ophthalmate, along with improvement in phosphatidylethanolamine to phosphatidylcholine (PE/PC) ratio.Conclusions: Vancomycin-mediated depletion of gram-positive bacteria negates the detrimental metabolic effect, dyslipidemia and IR, observed in iNOS-/- mice.

A novel volumetric velocity measurement method for small seeding tracers in large volumes

This paper presents the volumetric velocity measurement method of small seeding tracer with diameter 5µm ∼ 100µm for volumes of ≥ 500cm3. The size of seeding tracer is between helium-filled soap bubbles (HFSB) and di-ethyl-hexyl-sebacic acid ester(DEHS) droplets. The targeted measurement volume dimension is equivalent to the volume of HFSB, which will give a higher resolution of turbulence study. The relations between particle size, imaging and light intensity are formulated. The estimation of the imaging results is computed for the setup design. Finally, the methodology is demonstrated for turbulence velocity measurements in the jet flow, in which the velocities of averaged diameter 15µm air filled soap bubbles are measured in a volume of 7200cm3.

Biomaterial implants in the treatment of oncology: a review

In globally, cancer is a second leading disease next to cardiovascular diseases in non-communicable diseases, which affect the all ages, sex, social status, ethnicity and primary cause of illness related death. Traditionally, systemic delivery drug systems like chemotherapy via oral capsule, injections of nanoparticles/micro particles, immunotherapy and others, which can inhibit or halt the progression of tumors. The short half-life of drugs which cannot achieve the targeted dose level to the tumor site and not able to target desired cell and commonly produces the organ toxicity. Recently, researchers have been attempting to direct delivery agents for cancer therapy. One of the best methods is a local therapy system, which deliver the drug directly via implantable procedure and it’s achieved the maximum concentration of the desire drug at the tumor site, non-target systemic exposure and minimize the organ toxicity to the patients. Biomaterial implants are widely used in the local concurrent delivery of chemotherapy and anti-angiogenic agents, local delivery of poly-chemotherapy, gene therapy as an alternative to drug delivery, scaffolds for cancer immunotherapy and polymer-based composites of drug molecules. There are different types of polymers like poly anhydride poly [bis (p-carboxy-phenoxy) propane-sebacic acid] copolymer [p(CPP:SA)], fatty acid dimer-sebacic acid copolymer (FAD-SA), poly (lactic-co-glycolic acid) copolymer (PLGA), poly (ε-caprolactone) (PCL), poly (glycerol monostearate-co-caprolactone), alginate and silica, used in successively cancer therapy. In order to minimize the risk of unwanted side effect of different types of biomaterials implants, it’s biocompatible to reduce the ability to elicit the inflammatory effect to the implanted area or the site. Therefore, the key role of choosing the appropriate and biocompatible implants to particular therapy is an indispensable. This should be validated with respect to risk benefit ratio in case of cancers. Biomaterial based implant local delivery systems provide more versatile and tailorable approach to against treatment of different types of the cancer.