Multidisciplinary utilization of dimethyl sulfoxide: pharmacological, cellular, and molecular aspects

Robert E. Chapin, Jane Adams, Kim Boekelheide, L. Earl Gray Jr, Simon W. Hayward, Peter S.J. Lees, Barry S. McIntyre, Kenneth M. Portier, Teresa M. Schnorr, Sherry G. Selevan, John G. Vandenbergh, and Susan R. Woskie Pfizer, Inc., Groton, CT University of Massachusetts, Boston, MA Brown University, Providence, RI U.S. Environmental Protection Agency, Research Triangle Park, NC Vanderbilt University Medical Center, Nashville, TN Johns Hopkins University, Baltimore, MD Schering Plough Research Institute, Summit, NJ American Cancer Society, Atlanta, GA National Institute for Occupational Safety and Health, Cincinnati, OH U.S. Public Health Service (Ret), Silver Spring, MD North Carolina State University, Raleigh, NC University of Massachusetts, Lowell, MA

https://onlinelibrary.wiley.com/doi/pdf/10.1002/bdrb.20147

NTP‐CERHR expert panel report on the reproductive and developmental toxicity of bisphenol A

Purpose To study the toxicity and pharmacokinetic-pharmacodynamic profile of 17-allylamino, 17-demethoxygeldanamycin (17-AAG) and to recommend a dose for phase II trials. Patients and Methods This was a phase I study examining a once-weekly dosing schedule of 17-AAG. Thirty patients with advanced malignancies were treated. Results The highest dose level reached was 450 mg/m(2)/week. The dose-limiting toxicities (DLTs) encountered were grade 3 diarrhea in three patients (one at 320 mg/m(2)/week and two at 450 mg/m(2)/week) and grade 3 to 4 hepatotoxicity (AST/ALT) in one patient at 450 mg/m(2)/week. Two of nine DLTs were at the highest dose level. Two patients with metastatic melanoma had stable disease and were treated for 15 and 41 months, respectively. The dose versus area under the curve-relationship for 17-AAG was linear (r(2) = .71) over the dose range 10 to 450 mg/m(2)/week, with peak plasma concentrations of 8,998 mu g/L (standard deviation, 2,881) at the highest dose level. After the demonstration of pharmacodynamic changes in peripheral blood leukocytes, pre- and 24 hours post-treatment, tumor biopsies were performed and demonstrated target inhibition (c-RAF-1 inhibition in four of six patients, CDK4 depletion in eight of nine patients and HSP70 induction in eight of nine patients) at the dose levels 320 and 450 mg/m(2)/week. It was not possible to reproducibly demonstrate these changes in biopsies taken 5 days after treatment. Conclusion It has been possible to demonstrate that 17-AAG exhibits a tolerable toxicity profile with therapeutic plasma concentrations and target inhibition for 24 hours after treatment and some indications of clinical activity at the dose level 450 mg/m(2)/week. We recommend this dose for phase II clinical trials.

/pdf/phase-i-pharmacokinetic-and-pharmacodynamic-study-of-17-47bibycex0.pdf

Phase I Pharmacokinetic and Pharmacodynamic Study of 17-Allylamino, 17-Demethoxygeldanamycin in Patients With Advanced Malignancies

Bone marrow stromal cells (MSC), which represent a population of multipotential mesenchymal stem cells, have been reported to undergo rapid and robust transformation into neuron-like phenotypes in vitro following treatment with chemical induction medium including dimethyl sulfoxide (DMSO; Woodbury et al. [2002] J. Neurosci. Res. 96:908). In this study, we confirmed the ability of cultured rat MSC to undergo in vitro osteogenesis, chondrogenesis, and adipogenesis, demonstrating differentiation of these cells to three mesenchymal cell fates. We then evaluated the potential for in vitro neuronal differentiation of these MSC, finding that changes in morphology upon addition of the chemical induction medium were caused by rapid disruption of the actin cytoskeleton. Retraction of the cytoplasm left behind long processes, which, although strikingly resembling neurites, showed essentially no motility and no further elaboration during time-lapse studies. Similar neurite-like processes were induced by treating MSC with DMSO only or with actin filament-depolymerizing agents. Although process formation was accompanied by rapid expression of some neuronal and glial markers, the absence of other essential neuronal proteins pointed toward aberrantly induced gene expression rather than toward a sequence of gene expression as is required for neurogenesis. Moreover, rat dermal fibroblasts responded to neuronal induction by forming similar processes and expressing similar markers. These studies do not rule out the possibility that MSC can differentiate into neurons; however, we do want to caution that in vitro differentiation protocols may have unexpected, misleading effects. A dissection of molecular signaling and commitment events may be necessary to verify the ability of MSC transdifferentiation to neuronal lineages.

Reevaluation of in vitro differentiation protocols for bone marrow stromal cells: disruption of actin cytoskeleton induces rapid morphological changes and mimics neuronal phenotype.

https://flore.unifi.it/bitstream/2158/941731/1/JOURNAL%20OF%20FUNCTIONAL%20FOODS%202014%20-%201-s2.0-S1756464613003009.pdf

Nutraceuticals and dyslipidaemia: Beyond the common therapeutics

Lipopolysaccharides (LPS) are cell wall components of Gram-negative bacteria. These molecules behave as bacterial endotoxins and their release into the bloodstream is a determinant of the development of a wide range of pathologies. These amphipathic molecules can self-aggregate into supramolecular structures with different shapes and sizes. The formation of these structures occurs when the LPS concentration is higher than the apparent critical micelle concentration (CMC(a)). Light scattering spectroscopy (both static and dynamic) was used to directly characterize the aggregation process of LPS from Escherichia coli serotype 026:B6. The results point to a CMC(a) value of 14 microg mL(-1) and the existence of premicelle LPS oligomers below this concentration. Both structures were characterized in terms of molecular weight (5.5 x 10(6) and 16 x 10(6) g mol(-1) below and above the CMC(a), respectively), interaction with the aqueous environment, gyration radius (56 and 105 nm), hydrodynamic radius, (60 and 95 nm) and geometry of the supramolecular structures (nearly spherical). Our data indicates that future in vitro experiments should be carried out both below and above the CMC(a). The search for drugs that interact with the aggregates, and thus change the CMC(a) and condition LPS interactions in the bloodstream, could be a new way to prevent certain bacterial-endotoxin-related pathologies.

Evaluation of Lipopolysaccharide aggregation by light scattering spectroscopy

Purpose of the study: To determine the effects of acetylcholine and SpermineNONOate - a NO donor - on RBC membrane and oxygen carrying properties. Material and methods: Aliquots of venous blood from eleven heathy subjects were incubated with ACh 10 −3 M, ACh 10 −5 M, SpermineNONOate 10 −5 M and SpermineNONOate 10 −4 M. The follow- ing parameters were determined: erythrocyte aggregation and deformability, hematocrit, plasma pH, osmolality, K + ,N a + , Ca 2+ concentrations, hemoglobin, oxyhemoglobin, carboxihemoglobin and methemoglobin concentrations, oxygen saturation of hemoglobin, oxygen and carbon dioxide parcial pressures and p50. Results: In presence of ACh there is an increase of erythrocyte deformability, decrease of erythrocyte aggregation, plasma pH, K + and Na + concentration, increase of Ca 2+ con- centration and p50. In presence of SpermineNONOate there an increase of erythrocyte deformability, plasma pH, decrease of Na + and Ca 2+ concentration, increase of metHb concentration and decrease of p50. Conclusion: Acetylcholine and Spermi- neNONOate are able to induce changes on RBC membrane and oxygen carrying properties.

Effects of acetylcholine and spermineNONOate on erythrocyte hemorheologic and oxygen carrying properties.

Acetylcholine has been detected in human blood. Acetylcholine receptors and acetylcholinesterase are present in erythrocyte membranes. We tested the acetylcholine and choline effects on nitric oxide metabolites (NOx), namely nitrites and nitrates, and observed if they are dependent on interactions with muscarinic receptors and acetylcholinesterase. Human erythrocyte suspensions were incubated with acetylcholine and choline in the absence or presence of 10 microM atropine or 10 microM velnacrine maleate. The nitrite and nitrate concentrations were determined by the Griess method. Acetylcholine or choline increased NOx control concentrations (P <0.001). The nitrite concentrations decreased in the presence of atropine or velnacrine maleate (P <0.03). The nitrate concentrations only decreased when velnacrine maleate was incubated with acetylcholine or choline (10 microM, P <0.03). These results demonstrated that acetylcholine and choline modulate nitric oxide metabolites on erythrocytes and this effect is mediated by interactions with erythrocyte membrane muscarinic receptors and membrane enzyme acetylcholinesterase. A hypothesis for the signal transduction mechanism has been discussed for acetylcholinesterase and muscarinic receptor (M1) participation.

J. Martins-Silva

Papers

Multidisciplinary utilization of dimethyl sulfoxide: pharmacological, cellular, and molecular aspects

Evaluation of Lipopolysaccharide aggregation by light scattering spectroscopy

Effects of acetylcholine and spermineNONOate on erythrocyte hemorheologic and oxygen carrying properties.

Acetylcholine and choline effects on erythrocyte nitrite and nitrate levels.

Amperometric measurements of nitric oxide in erythrocytes.