Aims: Skin burns remain a noteworthy general medical issue throughout the world, as it boosts a condition of immuno-suppression. The present research aimed to evaluate the efficacy of Syzygium aromaticum extracts, silver sulphadiazine ointment, and different commercially available topical antibiotics against pathogenic bacteria, isolated from the skin of burn patients. Methodology and results: A total of 124 clinical pus samples were collected from the skin of burn patients, admitted to two different tertiary care burn units at Peshawar, Pakistan. From these pus samples, 6 bacterial isolates from burned skin (Staphylococcus epidermidis, Streptococcus, Klebsiella, Enterobacter, Bacillus and Pseudomonas spp.) were isolated, while 4 different bacterial isolates (Staphylococcus epidermidis, Staphylococcus aureus, Bacillus and Streptococcus spp.) were isolated from unburned skin via conventional culturing techniques. Further, antibacterial assays were performed to compare the efficacy of S. aromaticum extracts (methanolic and aqueous extract), silver sulphadiazine ointment, and different commercially available antibiotics against tested bacteria. It was observed that both methanolic and aqueous extracts of S. aromaticum were effective at all concentrations against all the tested bacteria. In addition, all the tested antibiotics expressed substantial activity against most of the bacterial isolates. While silver sulphadiazine ointment was observed to be less potent against isolated bacteria as compared to S. aromaticum extracts. Conclusion, significance and impact of study It was concluded that both aqueous and methanolic extracts of S. aromaticum were effective antimicrobial agents and could be used as an alternative to control bacterial infections of burn patients. This study would help to distinguish the risk factors of bacterial pathogenicity in burn patients and would also provide a guideline to utilize medicinal plants and their extracts to minimize the chances of antibiotic resistance phenomenon in burn patients.
Anti-Bacterial Agents ; Cell Membrane ; Oxidative Stress ; Permeability ; Plant Extracts ; Syzygium

The kidney collecting duct (CD) is a tubular segment of the kidney where the osmolality and final flow rate of urine are established, enabling urine concentration and body water homeostasis. Water reabsorption in the CD depends on the action of arginine vasopressin (AVP) and a transepithelial osmotic gradient between the luminal fluid and surrounding interstitium. AVP induces transcellular water reabsorption across CD principal cells through associated signaling pathways after binding to arginine vasopressin receptor 2 (AVPR2). This signaling cascade regulates the water channel protein aquaporin-2 (AQP2). AQP2 is exclusively localized in kidney connecting tubules and CDs. Specifically, AVP stimulates the intracellular translocation of AQP2-containing vesicles to the apical plasma membrane, increasing the osmotic water permeability of CD cells. Moreover, AVP induces transcription of the Aqp2 gene, increasing AQP2 protein abundance. This review provides new insights into the transcriptional regulation of the Aqp2 gene in the kidney CD with an overview of AVP and AQP2. It summarizes current therapeutic approaches for X-linked nephrogenic diabetes insipidus caused by AVPR2 gene mutations.
Aquaporin 2 ; Arginine Vasopressin ; Body Water ; Cell Membrane ; Diabetes Insipidus, Nephrogenic ; Gene Expression Regulation ; Homeostasis ; Kidney ; Kidney Tubules, Collecting ; Osmolar Concentration ; Permeability ; Phenobarbital ; Receptors, Vasopressin ; Water

Antifungal drug resistance is a significant clinical problem, and antifungal agents that can evade resistance are urgently needed. In infective niches, resistant organisms often co-existed with sensitive ones, or a subpopulation of antibiotic-susceptible organisms may evolve into resistant ones during antibiotic treatment and eventually dominate the whole population. In this study, we established a co-culture assay in which an azole-resistant Candida albicans strain was mixed with a susceptible strain labeled with green fluorescent protein to mimic in vivo conditions and screen for antifungal drugs. Fluconazole was used as a positive control to verify the validity of this co-culture assay. Five natural molecules exhibited antifungal activity against both susceptible and resistant C. albicans. Two of these compounds, retigeric acid B (RAB) and riccardin D (RD), preferentially inhibited C. albicans strains in which the efflux pump MDR1 was activated. This selectivity was attributed to greater intracellular accumulation of the drugs in the resistant strains. Changes in sterol and lipid compositions were observed in the resistant strains compared to the susceptible strain, and might increase cell permeability to RAB and RD. In addition, RAB and RD interfered with the sterol pathway, further aggregating the decrease in ergosterol in the sterol synthesis pathway in the MDR1-activated strains. Our findings here provide an alternative for combating resistant pathogenic fungi.
ATP-Binding Cassette Transporters ; genetics ; metabolism ; Antifungal Agents ; chemistry ; metabolism ; pharmacology ; Azoles ; pharmacology ; Biosynthetic Pathways ; drug effects ; genetics ; Candida albicans ; chemistry ; drug effects ; metabolism ; Cell Membrane ; chemistry ; metabolism ; Coculture Techniques ; Drug Resistance, Fungal ; drug effects ; Ergosterol ; metabolism ; Fungal Proteins ; genetics ; metabolism ; Lipids ; chemistry ; Molecular Structure ; Permeability ; Phenyl Ethers ; chemistry ; metabolism ; pharmacology ; Sterols ; chemistry ; metabolism ; Stilbenes ; chemistry ; metabolism ; pharmacology ; Triterpenes ; chemistry ; metabolism ; pharmacology

Cyclosporin A (CsA) does not only exert a toxic effect on kidney parenchymal cells, but also protects them against necrotic cell death by inhibiting opening of mitochondrial permeability transition pore. However, whether CsA plays a role in hydrogen peroxide-induced kidney proximal tubular cell death is currently unclear. In the present study, treatment with CsA further increased apoptosis and necrosis in HK-2 human kidney proximal tubule epithelial cells during exposure to hydrogen peroxide. In addition, hydrogen peroxide-induced p53 activation and BH3 interacting-domain death agonist (BID) expression were higher in CsA-treated cells than those in non-treated cells, whereas hydrogen peroxide-induced activation of mitogen-activated protein kinases including p38, c-Jun N-terminal kinase, and extracellular signal-regulated kinase and activation of protein kinase B were not significantly altered by treatment with CsA. In oxidant-antioxidant system, reactive oxygen species (ROS) production induced by hydrogen peroxide was further enhanced by treatment with CsA. However, expression levels of antioxidant enzymes including manganese superoxide dismutase, copper/zinc superoxide dismutase, and catalase were not altered by treatment with hydrogen peroxide or CsA. Treatment with CsA further enhanced mitochondrial membrane potential induced by exposure to hydrogen peroxide, although it did not alter endoplasmic reticulum stress based on expression of glucose-regulated protein 78 and 94. Taken together, these data suggest that CsA can aggravate hydrogen peroxide-induced cell death through p53 activation, BID expression, and ROS production.
Apoptosis ; Catalase ; Cell Death ; Cyclosporine ; Endoplasmic Reticulum Stress ; Epithelial Cells ; Humans ; Hydrogen Peroxide ; Hydrogen ; JNK Mitogen-Activated Protein Kinases ; Kidney ; Membrane Potential, Mitochondrial ; Mitogen-Activated Protein Kinases ; Necrosis ; Permeability ; Phosphotransferases ; Proto-Oncogene Proteins c-akt ; Reactive Oxygen Species ; Superoxide Dismutase

The development of low-frequency ultrasound imaging technology and the improvement of ultrasound contrast agent production technology mean that they play an increasingly important role in tumor therapy. The interaction between ultrasound and microbubbles and their biological effects can transfer and release microbubbles carrying genes and drugs to target tissues, mediate the apoptosis of tumor cells, and block the embolization of tumor microvasculature. With the optimization of ultrasound parameters, the development of targeted microbubbles, and the emergence of various composite probes with both diagnostic and therapeutic functions, low-frequency ultrasound combined with microbubble contrast agents will bring new hope for clinical tumor treatment.
Antineoplastic Agents/therapeutic use* ; Apoptosis ; Autophagy ; Cell Membrane Permeability ; Cell Proliferation ; Contrast Media/administration & dosage* ; Drug Delivery Systems ; Humans ; Microbubbles ; Microcirculation ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasms/therapy* ; Patient Safety ; Transfection ; Ultrasonic Therapy/methods*

The study is aimed to explore the effects of stress at different temperatures( 35,45,55 ℃) on membrane permeability,active oxygen metabolism and accumulation of effective substances in Lonicera japonica,and provide theoretical basis for reducing deterioration and revealing browning mechanism during postharvest processing of L. japonica. The cell membrane permeability( relative conductivity,MDA content),active oxygen metabolism( SOD,POD,PPO,CAT activity) and the accumulation of effective substances( chlorogenic acid,luteolin,neochlorogenic acid,cryptochlorogenic acid,3,5-dicaffeoylquinic acid,3,4-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid) of L. japonica were all studied by constant temperature drying method,and the results were analyzed by the SPSS 17. 0 statistical software. The results showed that MDA content in L. japonica was increased by 151. 14% at 35 ℃,SOD,POD,PPO and CAT activity were 29. 73%,42. 86%,105. 02% and 10. 74% higher than at 45 ℃,respectively. The order of effective substance content in L. japonica was 35 ℃ >45 ℃ >55 ℃. The changes of membrane permeability,activity of active oxygen metabolizing enzyme and accumulation of active components were significantly affected by different temperature stress. The indexes showed that physiological and active oxygen metabolizing enzyme activity of L. japonica was the highest under 35 ℃ stress,chlorogenic acid and luteolin were effectively accumulated,which provides basic data for solving browning problem in the postharvest processing of L. japonica.
Cell Membrane Permeability ; Chlorogenic Acid/metabolism* ; Hot Temperature ; Lonicera/physiology* ; Luteolin/metabolism* ; Oxygen/metabolism* ; Stress, Physiological

OBJECTIVES: As a first step to study the anticaries effect of ethanol alone, we investigated the effects of ethanol on the expression levels of the atpB gene and proton permeability of Streptococcus mutans in suspension cultures. METHODS: S. mutans UA159 was grown in brain heart infusion medium at either pH 4.8 or 6.8. The total extracted RNA was reverse-transcribed into cDNA using a Superscript™ First-Strand Synthesis System. The resulting cDNA and negative controls were amplified by ABI PRISM 7700 real-time PCR system with SYBR Green PCR Master Mix. For proton flux assay, bacterial suspensions were titrated to pH 4.6 with 0.5 M HCl, and then additional 0.5 M HCl was added to decrease the pH values by approximately 0.4 units. The subsequent increase in pH was monitored using a glass electrode. Ten percent (v/v) butanol was added to the suspensions at 80 min to disrupt the cell membrane. RESULTS: In a concentration-dependent manner, ethanol alone not only decreased the growth rate of S. mutans and the expression of the atpB gene but also increased the proton permeability at both pH 4.8 and 6.8. CONCLUSIONS: These findings suggest that ethanol has the potential for an anticaries ingredient. We believe that ethanol may be used together with fluoride and/or other cariostatic agents in order to develop better anticaries toothpastes and/or mouthrinses.
Brain ; Cariostatic Agents ; Cell Membrane ; DNA, Complementary ; Electrodes ; Ethanol ; Fluorides ; Gene Expression ; Glass ; Heart ; Hydrogen-Ion Concentration ; Permeability ; Polymerase Chain Reaction ; Protons ; Real-Time Polymerase Chain Reaction ; RNA ; Streptococcus mutans ; Streptococcus ; Suspensions ; Toothpastes

Vasicinone, a quinazoline alkaloid from Adhatoda vasica Nees. is well known for its bronchodilator activity. However its antiproliferative activities is yet to be elucidated. Here-in we investigated the anti-proliferative effect of vasicinone and its underlying mechanism against A549 lung carcinoma cells. The A549 cells upon treatment with various doses of vasicinone (10, 30, 50, 70 µM) for 72 h showed significant decrease in cell viability. Vasicinone treatment also showed DNA fragmentation, LDH leakage, and disruption of mitochondrial potential, and lower wound healing ability in A549 cells. The Annexin V/PI staining showed disrupted plasma membrane integrity and permeability of PI in treated cells. Moreover vasicinone treatment also lead to down regulation of Bcl-2, Fas death receptor and up regulation of PARP, BAD and cytochrome c, suggesting the anti-proliferative nature of vasicinone which mediated apoptosis through both Fas death receptors as well as Bcl-2 regulated signaling. Furthermore, our preliminary studies with vasicinone treatment also showed to lower the ROS levels in A549 cells and have potential free radical scavenging (DPPH, Hydroxyl) activity and ferric reducing power in cell free systems. Thus combining all, vasicinone may be used to develop a new therapeutic agent against oxidative stress induced lung cancer.
Apoptosis ; Cell Membrane ; Cell Survival ; Cell-Free System ; Cytochromes c ; DNA Fragmentation ; Down-Regulation ; Justicia ; Lung Neoplasms ; Lung* ; Oxidative Stress ; Permeability ; Receptors, Death Domain ; Up-Regulation ; Wound Healing

OBJECTIVETo investigate the effect of c-Met inhibitor cabozantinib (XL-184) in inhibiting Listeria monocytogenes (LM) from invading Caco-2 cells to reduce the cell injury.

METHODSThe cell invasion capacity of LM was assayed in Caco-2 cells incubated with different doses of XL-184 for different durations. Caco-2 cells incubated with XL-184 were seeded on the upper room of the transwell chamber, and the cell monolayer was exposed to LM infection followed by addition of horseradish peroxidase (HRP). The trans-epithelial electric resistance (TEER), HRP concentration and LM colony-forming unit (CFU) were measured in the cell monolayer. Fluorescent staining was used to evaluate the cell viability, and LDH release from the cells was examined to assess the changes in cell membrane permeability.

RESULTSXL-184 significantly decreased LM invasion rate in Caco-2 cells in a dose- and time-dependent manner (P=0.000), and this effect was enhanced by co-incubation of the cells with ampicillin (P<0.05). In the cell membrane permeability assay in the monolayer cells, XL-184 markedly inhibited LM-induced reduction of TEER (P<0.05) and significantly suppressed LM-induced enhancement of cell membrane permeability shown by reduced HRP concentration and LM count in the lower chamber (P=0.000). The cells infected with LM showed significantly lowered cell viability, which was rescued by XL-184 (P<0.01); XL-184 also dose-dependently reduced LDH release from the cells (P<0.05).

CONCLUSIONSXL-184 can suppress LM invasion in Caco-2 cells to reduce the cell injury, suggesting its value as a promising candidate agent for prevention and treatment of LM infections.

Anilides ; pharmacology ; Caco-2 Cells ; Cell Membrane Permeability ; drug effects ; Cell Survival ; Humans ; Listeria monocytogenes ; drug effects ; Pyridines ; pharmacology

OBJECTIVES: The aim of this research was to determine the pH-dependent changes in F-ATPase activity and proton fluxes in Streptococcus mutans (S. mutans) as induced by varying the concentration of fluoride ±10 mM (0.058% (v/v)) ethanol. METHODS: S. mutans UA159 was grown in Brain Heart Infusion medium at pH 4.8, 6.8, or 8.8. The F-ATPase assay was initiated by the addition of ATP, and stopped by adding 10% trichloroacetic acid. For the proton flux assay, bacterial suspensions were titrated to pH 4.6 with 0.5 M HCl, and then 0.5 M HCl was added to decrease the pH values in units of approximately 0.4 pH. The subsequent increase in pH was monitored using a glass electrode. To disrupt the cell membrane, 10% (v/v) butanol was added to the suspensions after 80 minutes. RESULTS: At all pH levels, fluoride ±10 mM ethanol not only decreased F-ATPase activity but also increased the proton permeability of S. mutans. The largest effects were observed at pH 4.8. Ethanol enhanced these effects only at pH 4.8. CONCLUSIONS: A very low concentration of ethanol enhanced the action of fluoride on F-ATPase activity and the proton permeability in S. mutans at acidic pH levels. We expect that low concentrations of ethanol may be used together with fluoride and/or other anticaries agents to develop more effective anticaries preparations.
Adenosine Triphosphate ; Brain ; Cell Membrane ; Electrodes ; Ethanol* ; Fluorides* ; Glass ; Heart ; Hydrogen-Ion Concentration ; Permeability* ; Protons* ; Streptococcus mutans* ; Streptococcus* ; Suspensions ; Trichloroacetic Acid