Concentration-dependent Profile Research Articles

Sesquiterpene-evoked phytochemical toxicity in PC12 neuronal cells reveals a variable degree of oxidative stress and alpha-tocopherol and glutathione-dependent protection

Phytochemicals are often promoted generally as antioxidants and demonstrate variable levels of reactive oxygen species (ROS) sequestration in vitro, which attributes to their neuroprotective bioactivity. Sesquiterpenes from cannabis and essential oils may demonstrate bifunctional properties towards cellular oxidative stress, possessing pro-oxidant activities by generating ROS or scavenging ROS directly. Sesquiterpenes can also oxidize forming sesquiterpene oxides, however the relative contribution they make to the bioactivity or cytotoxicity of complex botanical extracts more generally is unclear, while selected cannabis-prevalent terpenes such as β-caryophyllene may also activate cannabinoid receptors as part of their biological activity. In the present study, we investigated selected sesquiterpenes β-caryophyllene and humulene and their oxidized forms (β-caryophyllene oxide and zerumbone, respectively) against established antioxidants (ascorbic acid, α-tocopherol, and glutathione) and in the presence of cannabinoid receptor 1 and cannabinoid receptor 2 antagonists, to gain a better understanding of the molecular and cellular mechanisms of neuroprotection versus neurotoxicity in semi-differentiated rat neuronal phaeochromocytoma (PC12) cells. Our results demonstrate that the sesquiterpenes β-caryophyllene, humulene and zerumbone possess concentration-dependent neurotoxic effects in PC12 cells. Both β-caryophyllene- and humulene-evoked toxicity was unaffected by CB1 or CB2 receptor antagonism, demonstrating this occurred independently of cannabinoid receptors. Both glutathione and α-tocopherol were variably able to alleviate the concentration-dependent loss of PC12 cell viability from exposure to β-caryophyllene, humulene and zerumbone. During 4-hour exposure to sesquiterpenes only modest increases in ROS levels were noted in PC12 cells, with glutathione co-incubation significantly inhibiting intracellular ROS production. However, significant increases in ROS levels in PC12 cells were demonstrated during 24-hour incubation with either antioxidants or sesquiterpenes individually, and with additive toxicity exhibited in combination. Overall, the results highlight a concentration-dependent profile of sesquiterpene neurotoxicity independent of cannabinoid receptors and dissociated from the formation of reactive oxygen species as a marker or correlate to the loss of cell viability.

Adsorption of Chromium, Copper, Lead, Selenium, and Zinc ions into ecofriendly synthesized magnetic iron nanoparticles.

The iron nanoparticles (Fe-NPs) have been synthesized using an environmentally friendly and simple green synthesis method. This study aims to obtain an aqueous extract from natural material wastes for synthesizing Fe-NPs. The produced Fe-NPs were evaluated as adsorbents for removing Pb, Se, Cu, Zn, and Cr from aqueous solutions. The formation of Fe-NPs was observed on exposure of the aqueous extract to the ferrous chloride and ferric chloride solutions. The characterization of the synthesized Fe-NPs was carried out using different instrumental techniques. As a function of the initial metal ion concentration, contact time, and various doses, the removal of the heavy metal ions was investigated. The UV-Vis spectrum of Fe-NPs showed a peak at 386 nm, 386 nm, 400 nm, 420 nm, 210 nm, 215 nm, and 272 nm of banana, pomegranate, opuntia, orange, potato, and onion, respectively. The FT-IR spectra confirmed the attachment of bioactive molecules from plants on the Fe-NPs surface. The effective reduction of metal ions was greatly aided by the -OH functional groups. The functional groups were examined and responsible for adsorption process by nanoparticle powder sample, these peaks are 3400 cm-1, 2900 cm-1, 1600 cm-1,1000 cm-1, and 1550 cm-1. The magnetization measurements revealed superparamagnetic behavior in the produced iron oxide nanoparticles. Heavy metal ions uptake followed a time, dose, and initial concentration-dependent profile, with maximum removal efficiency at 45 min, 0.4 g, and 3.0 mg/L of metal concentration, respectively.

Biosynthesis of ZnO Nanoparticles Using the Aqueous Extract of Mucuna pruriens (utilis): Structural Characterization, and the Anticancer and Antioxidant Activities

A simple, green, and cost-effective synthesis of ZnO nanoparticles particles (NPs) using an extract of Mucuna pruriens utilis is reported. The nanoparticles were characterized by X-ray diffraction, UV–vis spectroscopy, SEM, and TEM measurements. XRD results showed diffraction patterns that are consistent with the hexagonal phase of the wurtzite ZnO structure. Spherical morphology with irregular size and particle distribution was confirmed by the microscopic characterization. The antioxidant activity of the nanoparticles showed a concentration-dependent profile with an IC50 of 4.10 µg mL− 1, which was quite lower than that of the standard ascorbic acid (4.72 µg mL− 1), and indicated a significant free radical scavenging activity of the nanomaterials. The cytotoxicity properties of the nanoparticles were evaluated against human cancer cell lines HeLa and HEK 293 by the MTT assay, and the anticancer drug (5-Fluorouracil) was used as a control. The results showed selective toxicity of the nanoparticles towards cancerous cell lines and non-toxicity to normal cells. The study provides a simple and non-toxic protocol for biosynthesis of ZnO nanoparticles with potential biomedical applications as anticancer and antioxidant agents. However, further studies are necessary to ascertain the biochemical reactions and mechanisms responsible for the antioxidant and anticancer activities.

Synthesis, characterization, and biological evaluation of some metal complexes containing N and S donor atoms

Eight new bipyridine adducts of Zn(II) and Ni(II) dithiocarbamate complexes have been prepared from the reaction of Zn(II) and Ni(II) complexes of N-ethyl-N-phenyldithiocarbamate and 4,4′ dipyridine, 4,4′-dimethyl-2,2′-bipyridine, 4,4′-dimethoxy-2,2′-bipyridine, 4,4′-ditertbutyl-2,2′-bipyridine. The resulting adducts were represented as [M(L1)2(DP)], [M(L1)2(DMeB)], [M(L1)2(DMxB)] and [M(L1)2(DTB)] respectively (where M = Zn or Ni). The structural configuration of these adducts was established using various spectroscopic techniques. The obtained data from the spectroscopic studies revealed a change in the tetrahedral configuration of the parent complexes to octahedral coordination in the adducts due to an additional metal-nitrogen bond. The formation of this bond was established by a considerable shift in the peaks around the metal-nitrogen (M−N) coordination upon the formation of the adducts. The study of biological properties including cytotoxicity, antioxidant, and anti-inflammatory properties were carried out. No noticeable trends were reported in all the studies despite the concentration-dependent profile shown in the assays and the structural similarities of the compounds. These adducts, nevertheless, showed good antioxidant activity in the DPPH assay, with IC50 ranging between 3.78 and 4.87 µg/mL which is better than the standard ascorbic acid (4.96 µg/mL). Also, in the cytotoxicity study, the adducts [Zn(L)2DMeBp] (1) and [Ni(L)2 DTBp] (6) showed the best activity against the mutating human cervical cancer (HeLa) and embryo kidney (HEK 293) cell lines and better than the standard 5-Flurouracil. The molecular docking studies further revealed that the adducts had a good binding affinity to the drug targets used for the study.

Atomic transport properties and liquid–liquid phase separation of Zn x Bi liquid monotectic alloys

Atomic transport properties specifically the shear viscosity and the diffusion coefficient for Zn x Bi liquid monotectic segregating alloys are theoretically investigated by using the Rice–Allnatt theory. The essential ingredient for the microscopic description of the metals and their alloys is the interionic interaction which in the present work is described by a widely used local pseudopotential. The temperature dependent behaviour of the above mentioned physical properties is also examined. The overall agreement of our calculated results with the available experimental data is found to be good for the full range of concentration. More interestingly, the temperature dependent results for the viscosity and the diffusion coefficient apparently exhibit a signature of liquid–liquid phase separation through a sudden bending in their concentration dependent profiles. Onset of this bending also provides information about the critical temperature and the critical concentration, and also provides a value for the critical exponent of liquid–liquid phase separation.

The concentration dependent SERS studies of a bioactive 4-chlorobenzylidene derivative: Experimental and DFT investigations

In this work, the vibrational spectroscopy of the compound (E)-4-(tert-butyl)-Nˈ-(4-chlorobenzylidene)benzyhydrazide (TCB), which was studied in conjunction with theoretical calculations, is applied for Surface Enhanced Raman Scattering (SERS) studies. The assignments of the vibrational modes were completed with Density Functional Theory (DFT)/B3LYP/6-311++G*. The information gathered is utilized to determine the chemical and electrical characteristics in addition to wavenumber measurements. The TCB molecule’s chemisorbed status on colloidal nanoparticles is confirmed by acquired red shift and decreased intensity behavior in the UV–VIS spectrum analysis. Additionally, the SERS spectra at varied concentrations and the chemisorption of the molecule with metal cluster were recorded and examined. Additionally, a silver cluster-based theoretical SERS model is put forth. The findings presented here will help in the construction of more dependable SERS sensors by thoroughly describing the concentration-dependent profile of analyte orientation.

Synthesis and Biological Evaluation of PEGylated MWO4 Nanoparticles as Sonodynamic AID Inhibitors in Treating Diffuse Large B-Cell Lymphoma.

Sonodynamic therapy (SDT) triggered by ultrasound (US) has attracted increasing attention owing to its ability to overcome critical limitations, including low tissue-penetration depth and phototoxicity in photodynamic therapy (PDT). Biogenic metal oxide nanoparticles (NPs) have been used as anti-cancer drugs due to their biocompatibility properties with most biological systems. Here, sonosensitizer MWO4-PEG NPs (M = Fe Mn Co Ni) were synthesized as inhibitors to activation-induced cytidine deaminase (AID), thus neutralizing the extensive carcinogenesis of AID in diffuse large B-cell lymphoma (DLBCL). The physiological properties of these nanomaterials were examined using transmission electron microscopy (TEM). The inhibition of NPs to AID was primarily identified by the affinity interaction prediction between reactive oxygen species (ROS) and AID through molecular dynamics and molecular docking technology. The cell apoptosis and ROS generation in US-triggered NPs treated DLBCL cells (with high levels of AID) were also detected to indicate the sonosensitivity and toxicity of MWO4-PEG NPs to DLBCL cells. The anti-lymphoma studies using DLBCL and AID-deficient DLBCL cell lines indicated a concentration-dependent profile. The synthesized MWO4-PEG NPs in this study manifested good sonodynamic inhibitory effects to AID and well treatment for AID-positive hematopoietic cancers.

Bipyridine adducts of Zn(II) and Ni(II) bis (N-methyl-N-phenyl dithiocarbamate): Synthesis, characterization, and biological applications

Substituted bipyridine adducts of Zn(II) and Ni(II) bis (N-methyl-N-phenyl dithiocarbamate) have been synthesized by the reaction of the dithiocarbamate complexes with 4, 4′ bipyridine, 4, 4′-dimethyl- 2,2 bipyridine, 4,4′-dimethoxy-2,2′-bipyridine, 4,4′-ditertbutyl-2,2′ bipyridine. The resulting adduct were represented as [M(L)2(B)], [M(L)2(DMeB)], [M(L)2(DMxB)] and [M(L)2(DTB)] respectively, (where M= Zn/Ni), and characterized using elemental analysis, various spectroscopic techniques and X-ray single crystallography. The spectroscopic data suggest octahedral coordination around the metal center due to the formation of a new M-N bond with the nitrogen of the Lewis bases. A considerable shift occurred in the peaks found in the vicinity of the M-N coordination upon the formation of the adducts. Single crystal data obatined for [Zn(L)2(B)] and [Zn(L)2(DMeB)] revealed a five and six geometry respectively around the Zn center in the adducts. Furthermore, the compounds were evaluated for cytotoxicity, antioxidant, and anti-inflammatory properties. Although all the biological studies conducted generally revealed a concentration-dependent profile, no noticeable trends were found even with the compounds that exhibited structural similarities. Nevertheless, the compounds showed good activity, with IC50 ranging from 0.0003 to 17.38 µg/mL, which was better than the standard 5-Fluorouracil (5 FU) used in the cytotoxicity study. The adducts had a better antioxidant property in DPPH assay than in all the used antioxidant assays. Upon evaluation in a Bovine serum albumin denaturation assay, they also possessed a better anti-inflammatory property than diclofenac.

Antileishmanial activity, cytotoxicity and cellular response of amphotericin B in combination with crotamine derived from Crotalus durissus terrificus venom using in vitro and in silico approaches

ObjectiveTo investigate the in vitro activity, synergism, cytotoxicity and cellular immunological response, as well as the molecular affinity between amphotericin B (AmB) and crotamine (CTA), derived from Crotalus durissus terrificus venom against Leishmania amazonensis. MethodsThis study performed the inhibition of promastigotes and amastigotes’ growth under different concentrations of the drug and pharmacological combinations (AmB + CTA) based on the Berimbaum method (synergism study). The lactate dehydrogenase (LDH) quantification method was used to determine the cytotoxicity of the drug and combinations employing four cell lines (J774, HepG2, VERO, and C2C12). Following, the levels of Tumour Necrose Factor-alpha (TNF-α) and Interleukin-12 (IL-12) cytokines, using enzyme-linked immunosorbent assay (ELISA) and nitrites, as an indirect measure of Nitric Oxide (NO), using the Griess reaction were assessed in the supernatants of infected macrophages. In silico approach (molecular docking and dynamics) and binding affinity (surface plasmon resonance) between the drug and toxin were also investigated. ResultsCTA enhanced AmB effect against promastigote and amastigote forms of L. amazonensis, decreased the drug toxicity in different cell lines and induced the production of important Th1-like cytokines and NO by infected macrophages. The pharmacological combination also displayed consistent molecular interactions with low energy of coupling and a concentration-dependent profile. ConclusionOur data suggest that this pharmacological approach is a promising alternative treatment against L. amazonensis infection due to the improved activity (synergistic effect) achieved against the parasites’ forms and to the decreased cytotoxic effect.

Resettable sweat-powered wearable electrochromic biosensor

Wearable biofuel cells (BFCs) are being widely studied as self-powered sensors to monitor one's physiological status non-invasively. However, the energy generated by such devices is often insufficient to power accompanying measurement read-out systems and communication protocols. Here, we report a reversible fully printed electrochromic self-powered sensor (ESPS) toward on-body sweat lactate monitoring. The device was realized by integrating a lactate-oxidizable BFC anode, oxygen-reducible BFC cathode, and a reversible Prussian blue (PB) electrochromic display on a single wearable patch. Through modification of the BFC electrodes, the formal potential of the PB display (0.14 V) could be positioned in between the potentials of the lactate oxidation (−0.05 V) and oxygen reduction (0.35 V). Based on this system, the ESPS enables reversible and robust color changes with distinct time- and concentration-dependent profiles. The results demonstrate that the self-powered device has practical on-body applications for continual lactate monitoring with several advantages, including facile reversibility, ease of fabrication, and naked-eye colorimetric read-out.

Biogenic Synthesis of CuO, ZnO, and CuO-ZnO Nanoparticles Using Leaf Extracts of Dovyalis caffra and Their Biological Properties.

Biogenic metal oxide nanoparticles (NPs) have emerged as a useful tool in biology due to their biocompatibility properties with most biological systems. In this study, we report the synthesis of copper oxide (CuO), zinc oxide (ZnO) nanoparticles (NPs), and their nanocomposite (CuO–ZnO) prepared using the phytochemical extracts from the leaves of Dovyalis caffra (kei apple). The physicochemical properties of these nanomaterials were established using some characterization techniques including X-ray diffraction analysis (XRD), ultraviolet-visible spectroscopy (UV-vis), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and energy-dispersive X-ray spectroscopy (EDX). The XRD result confirmed the presence of a monoclinic CuO (Tenorite), and a hexagonal ZnO (Zincite) nanoparticles phase, which were both confirmed in the CuO–ZnO composite. The electron microscopy of the CuO–ZnO, CuO, and ZnO NPs showed a mixture of nano-scale sizes and spherical/short-rod morphologies, with some agglomeration. In the constituent’s analysis (EDX), no unwanted peak was found, which showed the absence of impurities. Antioxidant properties of the nanoparticles was studied, which confirmed that CuO–ZnO nanocomposite exhibited better scavenging potential than the individual metal oxide nanoparticles (CuO, and ZnO), and ascorbic acid with respect to their minimum inhibitory concentration (IC50) values. Similarly, the in vitro anticancer studies using MCF7 breast cancer cell lines indicated a concentration-dependent profile with the CuO–ZnO nanocomposite having the best activity over the respective metal oxides, but slightly lower than the standard 5-Fluorouracil drug.

Assessments in early growth of corn seedlings after hausmanite (Mn3O4) nanoscale seed priming

Nanofertilizer application is becoming a sustainable alternative for plants micronutrients supply. Seed nutrient priming before seeding reduces non-target dispersion; although, applying nanofertilizer in correct concentration must be narrowly chosen to prevent germination and development issues. Here, we evaluated corn seedlings development and germination after seed priming with Mn3O4 nanoparticle (NP), Mn3O4 bulk and MnCl2. Sterile seeds were soaked for 8 hours in priming solutions of 0, 20, 40, 80 and 160 mg L−1 for each Mn sources. The seeds vigor and germination were evaluated after 7 days on germination paper. Root, shoot and total lengths were measured as well as root, shoot and total dry biomass. Compared to the control, the Mn3O4 NP and Mn3O4 bulk promoted beneficial effects. Mn3O4 NP seed-priming exhibited a concentration dependent profile in improving seedling growth, with greatest benefit around 20 mg L−1, providing higher germination, vigor, dry biomass and length than control and the other source tested. Particle size plays an important role in the reactivity of Mn3O4 NP. On the other hand, seeds primed with soluble source did not differ from the control. These findings support NP-seed priming as an alternative to delivery micronutrients.

Targeted uranium recovery from complex alloys using fluoride volatility

A single step separation of molybdenum from a multicomponent uranium alloy, 10 wt.% Mo in uranium metal, (U-10Mo)–Zr is explored using nitrogen trifluoride. The separation takes advantage of an unusually low temperature behavior of NF3. Exposure of the alloy to flowing NF3 between 100–200 °C caused surface area dependent, NF3 concentration, and temperature dependent thermal runaway that threatened the viability of the separation. Gravimetric methods were used to acquire temperature and NF3 concentration dependent profiles to understand the nature of the self-heating and to establish practical conditions for the separation. Scale-up to multigram quantities of the composite is described. In this effort, the use of larger product masses helped to divulge potential metal/MoF6 redox behavior of during the fluorination and acknowledgement of some unexpected chemistries as the metals evolved from the metallic state to their higher fluorides in the presence of what appears to be a catalytic or otherwise activated NF3-uranium metal surface.

Initial Development of Corn Seedlings after Seed Priming with Nanoscale Synthetic Zinc Oxide

Nanofertilizers are increasingly explored for sustainable micronutrient delivery in agriculture. Pre-treating seeds with nanofertilizers prior to planting (i.e., seed priming) reduces concerns about nanoparticle (NP) fertilizer non-target dispersion; however, priming formulations and concentrations must be carefully selected to avoid germination inhibition and toxicity. Here we investigate changes in corn seed germination and seedling development after seed priming with ZnO NPs, ZnO bulk and ZnCl2. To evaluate the effects sterile seeds were immersed in priming solutions of 0, 20, 40, 80, 160 mg L−1 Zn for the three Zn sources. Following an 8 h priming the seeds were evaluated for germination and vigor for 5 days on germination paper. Root and shoot lengths were measured as well as fresh and dry biomass. Compared to the control, the ZnO NP and ZnCl2 seed priming promoted beneficial effects. ZnO NP seed-priming exhibited a concentration dependent profile in improving seedling growth, with greatest benefit around 80 mg L−1, providing 17%, 25% and 12% higher values than control for germination, root length, and dry biomass production, respectively. In contrast, seeds primed with bulk ZnO did not differ from the control. These findings support NP-seed priming as an alternative to delivery of essential micronutrients, such as zinc, to corn seedlings.

Inhibitors of dual-specificity tyrosine phosphorylation-regulated kinases (DYRK) exert a strong anti-herpesviral activity

Infection with human cytomegalovirus (HCMV) is a serious medical problem, particularly in immunocompromised individuals and neonates. The success of (val)ganciclovir therapy is hampered by low drug compatibility and induction of viral resistance. A novel strategy of antiviral treatment is based on the exploitation of cell-directed signaling, e. g. pathways with a known relevance for carcinogenesis and tumor drug development. Here we describe a principle for putative antiviral drugs based on targeting dual-specificity tyrosine phosphorylation-regulated kinases (DYRKs). DYRKs constitute an evolutionarily conserved family of protein kinases with key roles in the control of cell proliferation and differentiation. Members of the DYRK family are capable of phosphorylating a number of substrate proteins, including regulators of the cell cycle, e.g. DYRK1B can induce cell cycle arrest, a critical step for the regulation of HCMV replication. Here we provide first evidence for a critical role of DYRKs during viral replication and the high antiviral potential of DYRK inhibitors (SC84227, SC97202 and SC97208, Harmine and AZ-191). Using established replication assays for laboratory and clinically relevant strains of HCMV, concentration-dependent profiles of inhibition were obtained. Mean inhibitory concentrations (EC50) of 0.98 ± 0.08 μM/SC84227, 0.60 ± 0.02 μM/SC97202, 6.26 ± 1.64 μM/SC97208, 0.71 ± 0.019 μM/Harmine and 0.63 ± 0.23 μM/AZ-191 were determined with HCMV strain AD169-GFP for the infection of primary human fibroblasts. A first analysis of the mode of antiviral action suggested a block of viral replication at the early-late stage of HCMV gene expression. Moreover, rhesus macaque cytomegalovirus (RhCMV), varicella-zoster virus (VZV) and herpes simplex virus (HSV-1) showed a similarly high sensitivity to these compounds. Thus, we conclude that DYRK signaling represents a promising target pathway for the development of novel anti-herpesviral strategies.

Liposomal Entrapment of 4-Nerolidylcatechol: Impact on Phospholipid Dynamics, Drug Stability and Bioactivity

Liposomes containing 4-nerolidylcatechol (4-NC), the major metabolite isolated from Pothomorphe umbellata, were obtained and characterized. Influence of liposomal encapsulation on chemical stability of 4-NC and on cytotoxicity profile of this drug was evaluated. Soybean phosphatidylcholine liposomes were prepared by lipid film hydration followed by extrusion. Entrapment efficiency for 4-NC was approximately 92%. Mean diameter of liposomes was 100 nm with a polydispersity index below 0.13. Liposomal 4-NC (L4-NC) and free drug (F4-NC) were submitted to forced degradation assays, monitored by HPLC. Photodegradation assay followed ICH Guidelines, using a photostability chamber equipped with both UV and white light sources. Liposomal encapsulation was able to markedly reduce 4-NC degradation rates under all the conditions tested. L4-NC showed a half-live approximately 15% higher than F4-NC under light exposure. After 72 hours, acid and base hydrolysis of F4-NC lead to 13 and 16% of degradation, respectively. However, no degradation was observed in L4-NC. EPR spectra of liposomal membrane showed that greatest changes in membrane properties were obtained when 5-doxyl stearic acid was used as the spin label, indicating a marked decrease in the fluidity of the bilayer. Following incubation with K562 cells, 4-NC showed a concentration-dependent cytotoxicity profile, while L4-NC exhibited a time and concentration-dependent profile, consistent with a controlled drug release system. F4-NC induced extensive hemolysis under isotonic conditions; conversely liposomal encapsulation protected erythrocytes from 4-NC induced lysis. Liposomal 4-NC resulted in a hemocompatibility and stable formulation, representing a viable drug delivery system to further investigate in vivo performances of 4-NC in pre clinical studies.

Drug repositioning: auranofin as a prospective antimicrobial agent for the treatment of severe staphylococcal infections.

Auranofin, (AF), a gold(I) complex in clinical use for the therapy of rheumatoid arthritis, is reported here to produce remarkable bactericidal effects in vitro against Staphylococcus sp. Noticeably, a similar antimicrobial action and potency are also noticed toward a few methicillin-resistant Staphylococcus aureus strains but not toward Escherichia coli. The time and concentration dependencies of the antimicrobial actions of AF have been characterized through recording time kill curves, and a concentration dependent profile highlighted. Overall, the present results point out that auranofin might be quickly and successfully repurposed for the treatment of severe bacterial infections due to resistant Staphylococci.

A Proposal of a Pharmacokinetic/pharmacodynamic (PK/PD) Index Map for Selecting an Optimal PK/PD Index from Conventional Indices (AUC/MIC, Cmax/MIC, and TAM) for Antibiotics

A pharmacokinetic/pharmacodynamic (PK/PD) analysis is important in antibiotic chemotherapy. Basically, the in vivo efficacy of antibiotics that exert concentration-dependent effects can be predicted using conventional PK/PD indices such as the ratio of the area under the curve to the minimum inhibitory concentration (AUC/MIC) and/or the ratio of the maximum plasma concentration to MIC (Cmax/MIC), whereas that of antibiotics with time-dependent effects can be determined using the period of time for which the drug concentration exceeds the MIC (time above MIC [TAM]). However, an optimal PK/PD index remains to be established for some antibiotics. Thus, a PK/PD model which describes the PK profile and effect of an antibiotic was developed, and the results obtained from this model were interpreted to form a PK/PD index map to assess the optimal PK/PD index for the antibiotic. The findings from the map were generally consistent with clinical outcomes even for the antibiotics which proved to be exceptions to the conventional classification. For example, AUC/MIC was an optimal index for azithromycin despite its time-dependent bactericidal activity, and Cmax/MIC was a poor index for arbekacin despite its concentration-dependent profile. Thus, the map would be useful for selecting the appropriate PK/PD index for an antibiotic.

Concentration- and time-dependent genotoxicity profiles of isoprene monoepoxides and diepoxide, and the cross-linking potential of isoprene diepoxide in cells

Isoprene, a possible carcinogen, is a petrochemical and a natural product being primarily produced by plants. It is biotransformed to 2-ethenyl-2-methyloxirane (IP-1,2-O) and 2-(1-methylethenyl)oxirane (IP-3,4-O), both of which can be further metabolized to 2-methyl-2,2′-bioxirane (MBO). MBO is mutagenic, but IP-1,2-O and IP-3,4-O are not. While IP-1,2-O has been reported being genotoxic, the genotoxicity of IP-3,4-O and MBO, and the cross-linking potential of MBO have not been examined. In the present study, we used the comet assay to investigate the concentration- and time-dependent genotoxicity profiles of the three metabolites and the cross-linking potential of MBO in human hepatocyte L02 cells. For the incubation time of 1 h, all metabolites showed positive concentration-dependent profiles with a potency rank order of IP-3,4-O > MBO > IP-1,2-O. In human hepatocellular carcinoma (HepG2) and human leukemia (HL60) cells, IP-3,4-O was still more potent in inducing DNA breaks than MBO at high concentrations (>200 μM), although at low concentrations (≤200 μM) IP-3,4-O exhibited slightly lower or similar potency to MBO. Interestingly, their time-dependent genotoxicity profiles (0.5–4 h) in L02 cells were different from each other: IP-1,2-O and MBO (200 μM) exhibited negative and positive profiles, respectively, with IP-3,4-O lying in between, namely, IP-3,4-O-caused DNA breaks did not change over the exposure time. Further experiments demonstrated that hydrolysis of IP-1,2-O contributed to the negative profile and MBO induced cross-links at high concentrations and long incubation times. Collectively, the results suggested that IP-3,4-O might play a significant role in the toxicity of isoprene.

Interaction of silver nanoparticles with proteins: A characteristic protein concentration dependent profile of SPR signal

Silver nanoparticles are finding increasing applications in biological systems, for example as antimicrobial agents and potential candidates for control drug release systems. In all such applications, silver nanoparticles interact with proteins and other biomolecules. Hence, the study of such interactions is of considerable importance. While BSA has been extensively used as a model protein for the study of interaction with the silver nanoparticles, studies using other proteins are rather limited. The interaction of silver nanoparticles with light leads to collective oscillation of the conducting electrons giving rise to surface plasmon resonance (SPR). Here, we have studied the protein concentration dependence of the SPR band profiles for a number of proteins. We found that for all the proteins, with increase in concentration, the SPR band intensity initially decreased, reaching minima and then increased again leading to a characteristic “dip and rise” pattern. Minimum point of the pattern appeared to be related to the isoelectric point of the proteins. Detailed dynamic light scattering and transmission electron microscopy studies revealed that the consistency of SPR profile was dependent on the average particle size and state of association of the silver nanoparticles with the change in the protein concentration. Fluorescence spectroscopic studies showed the binding constants of the proteins with the silver nanoparticles were in the nano molar range with more than one nanoparticle binding to protein molecule. Structural studies demonstrate that protein retains its native-like structure on the nanoparticle surface unless the molar ratio of silver nanoparticles to protein exceeds 10. Our study reveals that nature of the protein concentration dependent profile of SPR signal is a general phenomena and mostly independent of the size and structure of the proteins.