A randomized, proof-of-concept clinical trial on repurposing chlorcyclizine for the treatment of chronic hepatitis C.

Background & Aims: Chlorcyclizine HCl (CCZ) is a piperazine-class antihistamine with anti-hepatitis C virus (HCV) activity in vitro and in vivo. In a first-in-humans study for HCV, we evaluated the antiviral effects and safety of CCZ±ribavirin (RBV), characterized pharmacokinetic (PK) and viral kinetic (VK) patterns, and provide insights into CCZs mode of action against HCV.

Methods: Chronic HCV patients were randomized to CCZ (75 mg twice daily) or CCZ+weight-based RBV (1000/1200 mg daily) for 28 days.

Interaction of dipalmitoyl phosphatidylserine with ethanol: induction of an ordered gel phase at room temperature.

Using differential scanning calorimetry and small and wide-angle X-ray diffraction, we show that, unlike the saturated phosphatidylcholines, for which ethanol induces chain interdigitation in the gel state, and unlike natural phosphatidylserine in which the gel state is almost unaffected by the addition of ethanol, dipalmitoyl phosphatidylserine (DPPS) assumes an ordered structure after incubation at room temperature in the presence of as little as 5% (v/v) ethanol. In the liquid crystalline state, a progressive decrease in the interbilayer spacing is observed as a function of ethanol concentration, similar to what is found for natural phosphatidylserine (PS) and 1-palmitoyl-2-oleoyl-phosphatidylserine (POPS). The 0.

Phosphatidylcholine structure determines cholesterol solubility and lipid polymorphism.

In the present work, we demonstrate that phosphatidylcholine with (16:1)9 acyl chains undergoes polymorphic rearrangements in mixtures with 0.6-0.8 mol fraction cholesterol.

Phase separation of cholesterol from phosphatidylserine-cholesterol mixtures in the presence of the local anesthetic tetracaine.

Addition of the local anesthetic tetracaine (TTC) to multilamellar dispersions of natural phosphatidylserine (PS) causes changes in the thermotropic properties of the membrane, which can be detected by differential scanning calorimetry, and in the structure of the membrane as detected by X-ray diffraction. At molar ratio [PS]/ [TTC] approximately 8.5, the melting temperature of the phospholipid shifts downwards by approximately 2.

Novel properties of cholesterol-dioleoylphosphatidylcholine mixtures.

We have studied the properties of mixtures of cholesterol with dioleoylphosphatidylcholine (DOPC), and with several other phospholipids, including 1-stearoyl-2-oleoylphosphatidylcholine (SOPC) and dioleoleoylphosphatidylserine (DOPS), as a function of cholesterol molar fraction and of temperature. Mixtures of DOPC with a cholesterol molar fraction of 0.4 or greater display polymorphic behavior.

Phase separation of cholesterol and the interaction of ethanol with phosphatidylserine-cholesterol bilayer membranes.

Thermotropic and structural effects of ethanol on phosphatidylserine (PS) membranes containing up to 0.4 mol fraction cholesterol were investigated by differential scanning calorimetry, X-ray diffraction and fluorescence spectroscopy. It was found that in the presence of cholesterol, 10% (v/v) added ethanol depresses the melting temperature of the phospholipid by approximately 2 degrees C, similar to what was observed in the absence of cholesterol.

A new high-temperature transition of crystalline cholesterol in mixtures with phosphatidylserine.

Phosphatidylserine and cholesterol are two major components of the cytoplasmic leaflet of the plasma membrane. The arrangement of cholesterol is markedly affected by the presence of phosphatidylserine in model membranes. At relatively low mol fractions of cholesterol in phosphatidylserine, compared with other phospholipids, cholesterol crystallites are formed that exhibit both thermotropic phase transitions as well as diffraction of x-rays.

Cholesterol crystalline polymorphism and the solubility of cholesterol in phosphatidylserine.

There is a marked hysteresis between the heating and cooling polymorphic phase transition of anhydrous cholesterol. At a scan rate of 0.05 degrees C/min the difference in transition temperatures between heating and cooling scans is approximately 10 degrees C.

Effect of dehydroepiandrosterone on phosphatidylserine or phosphatidylcholine bilayers: DSC and X-ray diffraction study.

The effect of dehydroepiandrosterone (DHEA) on the thermotropic and structural properties of phosphatidylserine or phosphatidylcholine membranes was investigated by differential scanning calorimetry and X-ray diffraction. At molar fractions of sterol, X (sterol), less than approximately 0.2, DHEA interacts with both types of model membranes, depressing the melting temperature and reducing the enthalpy of melting.

The effect of ethanol on the structure of phosphatidylserine bilayers.

Thermotrophic and structural effects of ethanol on phosphatidylserine (PS) membranes were investigated by differential scanning calorimetry (DSC) and X-ray diffraction. It was found that up to 15% (v/v) added ethanol, there is little change in the melting temperature of the phospholipid and no change in the interbilayer (d) spacing in the gel phase, indicating that there is no interdigitation of the hydrocarbon chains. Above the melting temperature of the phospholipid, a large decrease of the d spacing, due primarily to a decrease in the thickness of the bilayer, was found.

Ligand-specific oligomerization of T-cell receptor molecules.

T cells initiate many immune responses through the interaction of their T-cell antigen receptors (TCR) with antigenic peptides bound to major histocompatibility complex (MHC) molecules. This interaction sends a biochemical signal into the T cell by a mechanism that is not clearly understood. We have used quasielastic light scattering (QELS) to show that, in the presence of MHC molecules bound to a full agonist peptide, TCR/peptide-MHC complexes oligomerize in solution to form supramolecular structures at concentrations near the dissociation constant of the binding reaction.

Phase behavior of mixtures of cholesterol and saturated phosphatidylglycerols.

The interaction of cholesterol with a series of saturated phosphatidylglycerols was investigated using differential scanning calorimetry and X-ray diffraction. We find that the miscibility of cholesterol in phosphatidylglycerol bilayers is lower than in the corresponding phosphatidylcholine bilayers and decreases with increasing acyl chain length of the phospholipid. The influence of the negative charge of the phosphatidylglycerol on cholesterol miscibility is discussed.

Cholesteryl hemisuccinate-cholesterol interaction: miscibility properties of the sterols.

Miscibility of cholesterol and cholesteryl hemisuccinate was investigated by differential scanning calorimetry and small-angle X-ray scattering. In an excess of cholesterol, above 2:1 mol ratio, phase separation takes place into a mixed phase and an almost pure cholesterol phase.

The effect of protons or calcium ions on the phase behavior of phosphatidylserine-cholesterol mixtures.

The influence of protons or calcium ions on the miscibility of cholesterol in phosphatidylserine has been examined using differential scanning calorimetry and X-ray diffraction. At pH 2.6, where the carboxyl group of the serine moiety is protonated, two endothermic transitions are observed in cholesterol-phosphatidylserine mixtures.

Stabilization of halophilic malate dehydrogenase.

Malate dehydrogenase from the extreme halophile, Halobacterium marismortui, is stable only in highly concentrated solutions of certain salts. Previous work has established that its physiological environment is saturated in KCl; it remains soluble is saturated NaCl or KCl solutions; also it unfolds in solutions containing less than 2.5 M-NaCl or -KCl, salt concentrations which are still relatively high.

Monomers, dimers, and minifilaments of vertebrate skeletal myosin in the presence of sodium pyrophosphate.

The self-assembly of myosin in the presence of sodium pyrophosphate was studied in the pH range between 7.0 and 8.5.

Macromolecular assemblies of myosin.

The self-assembly of myosin into filamentous structures is a highly cooperative and rapid process. Nevertheless, the presence of nonequivalent bonding interactions within the filament permits differential stabilization of several macromolecular assemblies of myosin under well-controlled ionic conditions in citrate/Tris buffer at pH 8.0.

Interaction of chromatin with NaCl and MgCl2. Solubility and binding studies, transition to and characterization of the higher-order structure.

Chicken erythrocyte chromatin containing histones H1 and H5 was carefully separated into a number of well-characterized fractions. A distinction could be made between chromatin insoluble in NaCl above about 80 mM, and chromatin soluble at all NaCl concentrations. Both chromatin forms were indistinguishable electrophoretically and both underwent the transition from the low salt "10 nm" coil to the "30 nm" higher-order structure solenoid by either raising the MgCl2 concentration to about 0.

Interaction and conformational changes of chromatin with divalent ions.

We have investigated the interaction of divalent ions with chromatin towards a closer understanding of the role of metal ions in the cell nucleus. The first row transition metal ion chlorides MnCl2, CoCl2, NiCl2 and CuCl2 lead to precipitation of chicken erythrocyte chromatin at a significantly lower concentration than the alkali earth metal chlorides MgCl2, CaCl2 and BaCl2. A similar distinction can be made for the compaction of chromatin to the "30 nm" solenoid higher order structure which occurs at lower MeCl2 concentration in the first group but at the same MeCl2 concentration within each group.

Isolation and physical studies of the intact supercoiled, the open circular and the linear forms of ColE1-plasmid DNA.

For the study of DNA conformations, conformational transitions, and DNA-protein interactions, covalently closed supercoiled ColE1-plasmid DNA has been purified from cultures of Escherichia coli harboring this plasmid and grown in the presence of chloramphenicol according to the method of D.B. Clewell [J.