Molecular modeling and UV-vis spectroscopic studies on the mechanism of action of reversed chloroquine (RCQ)

Reversed chloroquine (RCQ) is a multiple ligand compound active against chloroquine-sensitive and resistant falciparum malaria. It is composed by a 4-aminoquinoline moiety (like that present in chloroquine (CQ)) joined to imipramine (IMP), a modulating agent that also showed intrinsic antiplasmodial activity against Brazilian Plasmodium falciparum isolates resistant to CQ. Molecular modeling and ultraviolet-visible spectroscopy (UV-vis) studies strongly suggest that the interaction between RCQ and heme is predominant through the quinoline moiety in a mechanism of action similar to that observed for CQ. (C) 2010 Elsevier Ltd. All rights reserved.

Chloroquine inhibits Paracoccidioides brasiliensis survival within human monocytes by limiting the availability of intracellular iron

The mechanisms used by Paracoccidioides brasiliensis (Pb 18) to survive into monocytes are not clear. Cellular iron metabolism is of critical importance to the growth of several intracellular pathogens, including P. brasiliensis, whose capacity to multiply in mononuclear phagocytes is dependent on the availability of intracellular iron. Chloroquine, by virtue of its basic properties, has been shown to prevent release of iron from holotransferrin by raising endocytic and lysosomal pH, and thereby interfering with normal iron metabolism. Then, in view of this, we have studied the effects of CHLOR on P. brasiliensis multiplication in human monocytes and its effect on the murine paracoccidioidomycosis. CHLOR induced human monocytes to kill P. brasiliensis. The effect of CHLOR was reversed by FeNTA, an iron compound that is soluble at neutral to alkaline pH, but not by holotransferrin, which releases iron only in an acidic environment. CHLOR treatment of Pb 18-infected BALB/c mice significantly reduced the viable fungi recovery from lungs, during three different periods of evaluation, in a dose-dependent manner. This study demonstrates that iron is of critical importance to the survival of P. brasiliensis yeasts within human monocytes and the CHLOR treatment in vitro induces Pb 18 yeast-killing by monocytes by restricting the availability of intracellular iron. Besides, the CHLOR treatment in vivo significantly reduces the number of organisms in the lungs of Pb-infected mice protecting them from several infections. Thus, CHLOR was effective in the treatment of murine paracoccidioidomycosis, suggesting the potential use of this drug in patients' treatment.

Reduced Plasma Angiotensin Ii Levels Are Reversed By Hydroxyurea Treatment In Mice With Sickle Cell Disease.

Sickle cell disease (SCD) pathogenesis leads to recurrent vaso-occlusive and hemolytic processes, causing numerous clinical complications including renal damage. As vasoconstrictive mechanisms may be enhanced in SCD, due to endothelial dysfunction and vasoactive protein production, we aimed to determine whether the expression of proteins of the renin-angiotensin system (RAS) may be altered in an animal model of SCD. Plasma angiotensin II (Ang II) was measured in C57BL/6 (WT) mice and mice with SCD by ELISA, while quantitative PCR was used to compare the expressions of the genes encoding the angiotensin-II-receptors 1 and 2 (AT1R and AT2R) and the angiotensin-converting enzymes (ACE1 and ACE2) in the kidneys, hearts, livers and brains of mice. The effects of hydroxyurea (HU; 50-75mg/kg/day, 4weeks) treatment on these parameters were also determined. Plasma Ang II was significantly diminished in SCD mice, compared with WT mice, in association with decreased AT1R and ACE1 expressions in SCD mice kidneys. Treatment of SCD mice with HU reduced leukocyte and platelet counts and increased plasma Ang II to levels similar to those of WT mice. HU also increased AT1R and ACE2 gene expression in the kidney and heart. Results indicate an imbalanced RAS in an SCD mouse model; HU therapy may be able to restore some RAS parameters in these mice. Further investigations regarding Ang II production and the RAS in human SCD may be warranted, as such changes may reflect or contribute to renal damage and alterations in blood pressure.

Analytical solutions for Tokamak equilibria with reversed toroidal current

In tokamaks, an advanced plasma confinement regime has been investigated with a central hollow electric current with negative density which gives rise to non-nested magnetic surfaces. We present analytical solutions for the magnetohydrodynamic equilibria of this regime in terms of non-orthogonal toroidal polar coordinates. These solutions are obtained for large aspect ratio tokamaks and they are valid for any kind of reversed hollow current density profiles. The zero order solution of the poloidal magnetic flux function describes nested toroidal magnetic surfaces with a magnetic axis displaced due to the toroidal geometry. The first order correction introduces a poloidal field asymmetry and, consequently, magnetic islands arise around the zero order surface with null poloidal magnetic flux gradient. An analytic expression for the magnetic island width is deduced in terms of the equilibrium parameters. We give examples of the equilibrium plasma profiles and islands obtained for a class of current density profile. (C) 2011 American Institute of Physics. [doi: 10.1063/1.3624551]

Escape patterns of chaotic magnetic field lines in a tokamak with reversed magnetic shear and an ergodic limiter

The existence of a reversed magnetic shear in tokamaks improves the plasma confinement through the formation of internal transport barriers that reduce radial particle and heat transport. However, the transport poloidal profile is much influenced by the presence of chaotic magnetic field lines at the plasma edge caused by external perturbations. Contrary to many expectations, it has been observed that such a chaotic region does not uniformize heat and particle deposition on the inner tokamak wall. The deposition is characterized instead by structured patterns called magnetic footprints, here investigated for a nonmonotonic analytical plasma equilibrium perturbed by an ergodic limiter. The magnetic footprints appear due to the underlying mathematical skeleton of chaotic magnetic field lines determined by the manifold tangles. For the investigated edge safety factor ranges, these effects on the wall are associated with the field line stickiness and escape channels due to internal island chains near the flux surfaces. Comparisons between magnetic footprints and escape basins from different equilibrium and ergodic limiter characteristic parameters show that highly concentrated magnetic footprints can be avoided by properly choosing these parameters. (c) 2008 American Institute of Physics.

Analysis of Single-Nucleotide Polymorphisms in the crt-o and mdr1 Genes of Plasmodium vivax among Chloroquine-Resistant Isolates from the Brazilian Amazon Region

Plasmodium vivax parasites with chloroquine resistance (CQR) are already circulating in the Brazilian Amazon. Complete single-nucleotide polymorphism (SNP) analyses of coding and noncoding sequences of the pvmdr1 and pvcrt-o genes revealed no associations with CQR, even if some mutations had not been randomly selected. In addition, striking differences in the topologies and numbers of SNPs in these transporter genes between P. vivax and P. falciparum reinforce the idea that mechanisms other than mutations may explain this virulent phenotype in P. vivax.

Development of a Stability-Indicating LC Assay Method for Determination of Chloroquine

A simple and rapid development of a stability-indicating LC method for determination of chloroquine diphosphate in the presence of its hydrolysis, oxidative and photolysis degradation products is described. Stress testing showed that chloroquine diphosphate was degraded under basic conditions and by photolytic treatment but was stable under the other stress conditions investigated. Separation of the drug from its degradation products was achieved with a Nova Pack C18 column, 0.01 M PIC B7 and acetonitrile (40:60 v/v) pH 3.6, as mobile phase. Response was linear over the range 0.08-5.70 mu g mL(-1) (r = 0.996), with limits of detection and quantification (LOD and LOQ) of 0.17 and 0.35 mu g mL(-1), respectively.

Enantioselective determination of chloroquine and its n-dealkylated metabolites in plasma using liquid-phase microextraction and LC-MS

A selective method using three-phase liquid-phase microextraction (LPME) in conjunction with LC-MS-MS was devised for the enantioselective determination of chloroquine and its n-dealkylated metabolites in plasma samples. After alkalinization of the samples, the analytes were extracted into n-octanol immobilized in the pores of a polypropylene hollow fiber membrane and back extracted into the acidic acceptor phase (0.1 M TFA) filled into the lumen of the hollow fiber. Following LPME, the analytes were resolved on a Chirobiotic V column using methanol/ACN/glacial aceti acid/diethylamine (90:10:0.5:0.5 by volume) as the mobile phase. The MS detection was carried out using multiple reaction monitoring with ESI in the positive ion mode. The optimized LPME method yielded extraction recoveries ranging from 28 to 66%. The method was linear over 5 - 500 ng/mL and precision (RSD) and accuracy (relative error) values were below 15% for all analytes. The developed method was applied to the determination of the analytes in rat plasma samples after oral administration of the racemic drug.

Chloroquine - Induced Hair Depigmentation

A case of hair depigmentation induced by chloroquine diphosphate subacute overdosage in an 11-year-old patient with dermatomyositis is presented. Normal coloured hair growth occurred after normalisation of chloroquine dosage. A discussion on possible pathomechanisms of this phenomenon is made based on experimental data and previously reported patients with the same condition. Lupus (2009) 18, 264-266.

Ductus venosus Doppler and postnatal outcomes in fetuses with absent or reversed end-diastolic flow in the umbilical arteries

Objective: To evaluate the relationship between ductus venosus Doppler findings on the day of delivery and postnatal outcomes in pregnancies with absent or reversed end-diastolic (ARED) flow in the umbilical arteries. Study design: Postnatal outcomes of 103 newborns of pregnancies with a diagnosis of ARED flow on Doppler velocimetry of the umbilical arteries were analyzed retrospectively between January 1997 and December 2004. Single pregnancies and fetuses without malformations were included. The cases were divided into two groups according to the flow during atrial contraction (a-wave) in the ductus venosus on the day of delivery: group A, 20 cases with absent or reversed flow in the ductus venosus and group B, 83 cases with positive flow. The results were analyzed statistically using the chi-square test, Fisher`s exact test and the Mann-Whitney U test with the level of significance set at 5%. Results: All newborns were delivered by cesarean section. Gestational age was similar in the two groups (group A: 30 weeks and group B: 30.9 weeks, P = 0.23). Absent or reversed ductus venosus flow was associated with the following adverse postnatal outcomes: lower birthweight (P < 0.001), lower Apgar scores in the first (P = 0.001) and fifth minute (P = 0.001), a higher frequency of orotracheal intubation (P = 0.001) and pH at birth less than 7.20 (P < 0.001), pulmonary hemorrhage (P = 0.03), thrombocytopenia (P = 0.02), hypoglycemia (P = 0.01), intracranial hemorrhage (P = 0.02), and postnatal death (P = 0.007). Conclusion: The study of ductus venosus flow may provide additional information regarding the best time for interruption of pregnancies with ARED flow in the umbilical arteries characterized by extreme prematurity. (C) 2008 Elsevier Ireland Ltd. All rights reserved.

Compromised Bond Strength after Root Dentin Deproteinization Reversed with Ascorbic Acid

Introduction: The present study evaluated the effect of a reducing agent on the bond strength of deproteinized root canal dentin surfaces when using a self-adhesive versus dual-cured cement. Regional differences were also evaluated. Methods: A total of 45 bovine incisor roots were divided into 3 groups: irrigation with physiologic solution (control), 10-minute deproteinization with 5% NaOCl, and 10-minute deproteinization with 5% NaOCl followed by 10 minutes of 10% ascorbic acid. Fiber posts were cemented with either RelyX 0100 or RelyX ARC (with SingleBond 2 or Clearfil SE Bond). The push-out bond strength was evaluated after 24 hours of storage. Data were submitted to three-way analyses of variance and Dunnett 13 tests (alpha = 0.05). Results: No differences between cements were observed within the testing conditions, regardless of the adhesive (P < .05). Deproteinization reduced bond strengths. Subsequent treatment with ascorbic acid was capable of reversing bond strength value changes to levels similar to those of controls. Regional radicular differences were also found, where coronal > middle > apical. Conclusions: The reducing agent was capable. of reversing the effect of dentin deproteinization, and RelyX U100 behaved similarly to RelyX ARC. (J Endod 2010;36:130-134)

Effects of accentuated and reversed curve of Spee on apical root resorption

Introduction: In this study, we evaluated the influence of intrusion mechanics with accentuated and reversed curve of Spee on root resorption of the maxillary and mandibular incisors. Methods: A sample of 60 patients with Class I and Class II Division 1 malocclusions having nonextraction treatment was divided into 2 groups with the following characteristics: group 1 comprised 30 deepbite patients, treated with accentuated and reversed curve of Spee intrusion mechanics, with an initial mean age of 12.8 +/- 1.23 years (range, 10.01-15.32 years), and group 2 comprised 30 patients with normal overbite treated without intrusion mechanics, with an initial mean age of 12.87 +/- 1.43 years ( range, 10.02-15.36 years). Pretreatment and posttreatment periapical radiographs were used to evaluate root resorption. The groups were compared by using the Mann-Whitney U test. Correlation between root resorption and tooth movement was investigated with the Spearman correlation coefficient. Results: The deepbite group treated with accentuated and reversed curve of Spee had statistically greater root resorption ( 1.87) than the normal overbite group ( 1.54), at P=.017. Changes in overbite and vertical displacements of the maxillary central incisor apices had significant correlations to root resorption ( r = 0.30, P =.019; r = 0.27, P =.037, respectively). Conclusions: Accentuating and reversing the curve of Spee in the archwires to correct deep overbite causes more root resorption than nonintrusive mechanics.

Short report: Molecular evaluation of the efficacy of chloroquine treatment of uncomplicated Plasmodium falciparum malaria in East Timor

The efficacy of chloroquine treatment of uncomplicated Plasmodium falciparum malaria in East Timor was investigated via molecular tools. Genotyping of the polymorphic markers msp1 and msp2 was performed to investigate the number and type of parasite alleles in pre- and posttreatment blood samples collected from 48 patients. Patients were infected with a minimum of 8 msp1 and 14 msp2 allelic types of parasite, and 43% of the patients had more than one allelic type before treatment. The genotyping also revealed that 66.7% of the patients were infected with at least one identical allelic type of parasite before and after treatment and therefore were likely to have experienced recrudescence. All parasites in pre- and posttreatment blood samples carried the K76T mutation in pfcrt, regardless of the clinical response to chloroquine. The sequence polymorphism patterns in pfcrt in the majority of parasites examined were identical to those observed in Bougainville, Papua New Guinea.

Chloroquine or sulfadoxine-pyrimethamine for the treatment of uncomplicated, Plasmodium falciparum malaria during an epidemic in Central Java, Indonesia

A recent malaria epidemic in the Menoreh Hills of Central Java has increased concern about the re-emergence of endemic malaria on Java, which threatens the island's 120 million residents. A 28-day, in-vivo test of the efficacy of treatment of malaria with antimalarial drugs was conducted among 16 7 villagers in the Menoreh Hills. The treatments investigated, chloroquine (CQ) and sulfadoxine pyrimethamine (SP), constitute, respectively, the first- and second-line treatments for uncomplicated malaria in Indonesia. The prevalence of malaria among 1389 residents screened prior to enrollment was 33%. Treatment outcomes were assessed by microscopical diagnoses, PCR-based confirmation of the diagnoses, measurement of the whole-blood concentrations of CQ and desethylchloroquine (DCQ), and identification of the Plasmodium falciparum genotypes. The 28-day cumulative incidences of therapeutic failure for CQ and SP were, respectively, 47% (N = 36) and 22% (N = 50) in the treatment of P. falciparum, and 18% (N = 77) and 67% (N = 6) in the treatment of P. vivax. Chloroquine was thus an ineffective therapy for P. falciparum malaria, and the presence of CQ-resistant P. vivax and SP-resistant P. falciparum will further compromise efforts to control resurgent malaria on Java.