T‐cell CD38 expression in B‐chronic lymphocytic leukaemia

B‐cell chronic lymphocytic leukaemia (B‐CLL) is a heterogeneous disease with some patients having an indolent course never needs treatment, while others having rapidly progressive one requires intensive treatment. In recent decades, numerous prognostic markers, such as immunoglobulin variable region heavy‐chain (IgVH) mutational status, ZAP‐70 and the expression of CD38 on leukaemic cells were introduced to screen for patients likely to have progressive course of B‐CLL bearing the potential to facilitate risk‐adapted treatment strategies. In B‐CLL, T cell function is shown to be dysregulated. CD38 has been demonstrated to be an important transmembrane signalling molecule of T cell with a direct effect on its function. The present study was conducted to analyse CD38 expression on T cells by flow cytometry to evaluate its impact on the clinical course of 88 unselected B‐CLL patients and correlate it with other risk factors. CD38 expression level on T cells was shown to predict the clinical course of B‐CLL in male patients but not in female patients. Male patients showed CD38 expression on T cells in a stage‐dependent manner, in contrast to female patients who showed higher expression irrespective to clinical staging. CD38 expression on T cells negatively interacted with treatment‐free survival in male patients. Multivariate analysis revealed that CD38 expression level on T cells is an independent prognostic factor in B‐CLL male patients. Simultaneous evaluation of CD38 expression on both B‐CLL cells and T cells allowed predicting male patient groups with the most favourable prognosis as well as those with the worst. Copyright © 2009 John Wiley & Sons, Ltd.     

Glutathione S‐Transferase M1 and T1 Gene Polymorphisms and the Outcome of Chronic Hepatitis C Virus Infection in Egyptian Patients

We analysed the distribution of GSTM1 and GSTT1 gene polymorphisms in Egyptian patients with chronic hepatitis C, and investigated their relationship to the clinical outcome of chronic hepatitis C virus (HCV) infection. This study included 169 patients with chronic HCV infection and 145 healthy and matched controls.GSTM1 and GSTT1 polymorphisms were genotyped by multiplex polymerase chain reaction. Individual GSTM1 null and GSTT1 null genotypes were more frequent in patients versus control subjects [OR, 4 (95% CI, 2.5–6.4); P ? 0.001] and [OR, 1.7 (95% CI, 1.1–2.6); P = 0.025], respectively. The patient group showed a higher frequency of the combined GSTM1/GSTT1 double‐null genotype than the control group [OR, 1.8 (95% CI, 1.1–2.9); P = 0.016]. The distribution frequencies of the combined GSTM1/GSTT1 double‐null genotype were significantly different [OR, 0.5 (95% CI, 0.25–0.99); P = 0.049] between F0–F3 and F4. There were no significant differences between the two groups with regard to other genotypes. The combined GSTM1/GSTT1 double‐null genotype was significantly increased in Child‐Pugh C patients in comparison to Child‐Pugh A+B (P = 0.02). There was no significant difference between different classes with regard to other genotypes. In conclusion, we identified an association between the combined GSTM1/GSTT1 double‐null genotype and advanced liver fibrosis and outcome of chronic HCV infection in Egyptian patients.     

Co-administration of ketoconazole and tacrolimus therapy: a transplanted rat model

Background/aim The aim of this work is to study the effect of addition of ketoconazole to experimental kidney transplanted rat treated with tacrolimus and precludes the percentage of tacrolimus dose reduction. Material and methods The material of this work included 60 male Sprague Dawely rats subjected to renal allotransplantation. They were equally divided into five groups: Group I: served as control group, Group II: received FK506 3.2 mg/kg/bw, Group III: received FK506 2 mg/kg/bw, Group IV: received FK506 1 mg/kg/bw, Group V: received FK506 1 mg/kg/bw plus ketoconazole 20 mg/kg/day. FK506 trough level and laboratory investigations were determined at 0, 3, 7, 10, 14, and 27 days post-transplantation. Results In all groups loss of body weight was observed at day 27 after treatment compared to that before transplantation. Serum creatinine significantly increased at day 27 compared to the basal level in groups treated with 1.0 mg and 3.2 mg FK506 (1.80 ± 0.50 vs. 0.39 ± 0.06 P = 0.001) and (1.03 ± 0.26 vs. 0.50 ± 0.07 P = 0.001) respectively. While for 2.0 mg or 1.0 mg plus keto groups, no significant differences in serum creatinine levels over time (0.56 ± 0.22 vs. 0.44 ± 0.10 P = 0.106) and (0.55 ± 0.30 vs. 0.42 ± 0.08 P = 0.160) were observed. Conclusion Concomitant administration of ketoconazole and FK506 is safe and results in increase blood trough level concentration of FK506 with 50% dose reduction in transplanted rat model.     

Synthesis, in vitro characterization, and radiolabeling of N,N-dimethyl-2-(2'-amino-4'-substituted-phenylthio)benzylamines: potential candidates as selective serotonin transporter radioligands

A series of N,N-dimethylated and N-monomethylated analogues of N,N-dimethyl-2-(2'-amino-4'-iodophenylthio)benzylamine substituted at the 4'-phenyl position have been prepared and evaluated in vitro for serotonin transporter (SERT) selectivity. Several derivatives were prepared where the 4'-position was either unsubstituted 13 and 33a or substituted with methyl 14a and 33b, ethenyl 14b and 34, ethyl 16 and 35, hydroxymethyl 20 and 41, hydroxyethyl 22, fluoroethyl 23, hydroxypropyl 27, and fluoropropyl 28. Competition binding in cells stably expressing the transfected human SERT, dopamine transporter (DAT), and norepinephrine transporter (NET) using [(3)H]citalopram, [(3)H]WIN 35,428 or [(125)I]RTI-55, and [(3)H]nisoxetine, respectively, demonstrated the following order of SERT affinity (K(i) (nM)): 14a (0.25) > 16 (0.49) > 20 (0.57) > 14b (1.12) > 13 (1.59) > 33b (1.94) = 35 (2.04) > 23 (8.50) = 28 (8.55) > 41 (15.11) > 22 (51) > 33a (83.43) > 27 (92). The K(i) values revealed that most of these derivatives displayed a high affinity for the SERT and a high selectivity over the DAT and NET. Moreover, substitution at the 4'-position of the dimethylated and monomethylated benzylamines differently influenced SERT binding: (i) the dimethylated benzylamines exhibited higher SERT affinity than the monomethylated ones, (ii) alkyl, alkenyl, or hydroxymethyl functions at the 4'-position afford compounds with high SERT affinity, and (iii) omega-hydroxy and fluoro-substituted ethyl and propyl groups at the 4'-position decrease the SERT affinity. From this series, the dimethylated derivatives 13, 14a, 14b, 16, and 20 were radiolabeled with carbon-11 and their log P(7.4) was calculated as a measure of their potential brain penetrance as positron emission tomography SERT imaging agents.     

Co-administration of Ketoconazole and Tacrolimus Therapy: A Transplanted Rat Model

Background/Aim: The aim of this work is to study the safety and the effect of addition of ketoconazole to experimental kidney transplanted rat treated with tacrolimus and predicts the percentage of tacrolimus dose reduction. Material and Methods: The material of this work included 60 male Sprague Dawely rats subjected to renal allotransplantation. They were equally divided into five groups: Group I: served as control group, Group II: received FK506 3.2 mg/kg/bw, Group III: received FK506 2 mg/kg/bw, Group IV: received FK506 1 mg/kg/bw, Group V: received FK506 1 mg/kg/bw plus Ketoconazole 20 mg/kg/day. FK506 trough level and laboratory investigations were determined at 0, 3, 7, 10, 14, and 27 days post-transplantation. Results: In all groups loss of body weight was observed at day 27 after treatment compared to that before transplantation. Serum creatinine significantly increased at day 27 compared to the basal level in groups treated with 1.0 and 3.2 mg FK506 (1.80 ± 0.50 versus 0.39 ± 0.06 P = 0.001) and (1.03 ± 0.26 versus 0.50 ± 0.07 P = 0.001) respectively, while for 2.0 mg or 1.0 mg plus keto groups, no significant differences in serum creatinine levels over time (0.56 ± 0.22 versus 0.44 ± 0.10 P = 0.106) and (0.55 ± 0.30 versus 0.42 ± 0.08 P=0.160) were observed. Conclusion: Concomitant administration of Ketoconazole and FK506 in transplanted rat model is safe and results in increase of blood trough level concentration of FK506 with 50% reduction of its dose.     

Carbon-11 HOMADAM: a novel PET radiotracer for imaging serotonin transporters

Carbon-11-labeled N,N-dimethyl-2-(2'-amino-4'-hydroxymethylphenylthio)benzylamine (HOMADAM) was synthesized as a new serotonin transporter (SERT) imaging agent. METHODS: Carbon-11 was introduced into HOMADAM by preparation of N-methyl-2-(2'-amino-4'-hydroxymethylphenylthio)benzylamine followed by alkylation with carbon-11 iodomethane. Binding affinities of HOMADAM and the radiolabeling substrate, N-methyl-2-(2'-amino-4'-hydroxymethylphenylthio)benzylamine, were determined in cDNA transfected cells expressing human SERT, dopamine transporters (DAT) and norepinephrine transporters NET using [3H]citalopram, [(125)I]RTI-55 and [3H]nisoxetine, respectively. MicroPET brain imaging was performed in monkeys. Arterial plasma metabolites of HOMADAM were analyzed in a rhesus monkey by high-performance liquid chromatography (HPLC). RESULTS: HOMADAM displayed high affinity for the SERT (Ki = 0.6 nM). N-methyl-2-(2'-amino-4'-hydroxymethylphenylthio)benzylamine displayed moderate affinity for the SERT (Ki = 15.11 nM). The affinities of HOMADAM for the DAT and NET were 2000- and 253-fold lower, respectively, than for the SERT. [11C]HOMADAM was prepared from [11C]iodomethane in approximately 25% radiochemical yield (decay-corrected to end of bombardment). MicroPET brain imaging studies in monkeys demonstrated that [11C]HOMADAM uptake was selectively localized in the midbrain, thalamus, pons, caudate, putamen and medulla. The midbrain-to-cerebellum, pons-to-cerebellum, thalamus-to-cerebellum and putamen-to-cerebellum ratios at 85 min were 4.2, 2.8, 2.3 and 2.0, respectively. HOMADAM binding achieved quasi-equilibrium at 45 min. Radioactivity in the SERT-rich regions of monkey brain was displaceable with R,S-citalopram. Radioactivity in the DAT-rich regions of monkey brain was not displaceable with the DAT ligand RTI-113. Radioactivity in the SERT-rich regions of monkey brain was displaceable with the R,S-reboxetine, a NET ligand with a high nanomolar affinity for SERT. Arterial plasma metabolites of HOMADAM were analyzed in a rhesus monkey by HPLC and displayed a single peak that corresponded to unmetabolized HOMADAM. CONCLUSION: HOMADAM is an excellent candidate for PET primate imaging of brain SERTs.     

Synthesis, radiosynthesis, and biological evaluation of carbon-11 labeled 2beta-carbomethoxy-3beta-(3'-((Z)-2-haloethenyl)phenyl)nortropanes: candidate radioligands for in vivo imaging of the serotonin transporter with positron emission tomography

2beta-carbomethoxy-3beta-(3'-((Z)-2-iodoethenyl)phenyl)nortropane (mZIENT, 1) and 2beta-carbomethoxy-3beta-(3'-((Z)-2-bromoethenyl)phenyl)nortropane (mZBrENT, 2) were synthesized and evaluated for binding to the human serotonin, dopamine, and norepinephrine transporters (SERT, DAT, and NET, respectively) using transfected cells. Both 1 and 2 have a high affinity for the SERT (Ki=0.2 nM) and are approximately 160 times more selective for the SERT than the DAT. Compound 2 has a significantly higher affinity for the NET than 1, and this may be a result of the different size and electronegativity of the halogen atoms. MicroPET imaging in nonhuman primates with [11C]1 and [11C]2 demonstrated that both tracers behave similarly in vivo with high uptake being observed in the SERT-rich brain regions and peak uptake being achieved in about 55 min postinjection. Chase studies with citalopram and methylphenidate demonstrated that this uptake is the result of preferential binding to the SERT.     

Therapeutic plasma exchange a potential strategy for patients with advanced heart failure

Background: Previous reports had emphasized the importance of humoral immunity in heart failure in humans, primarily determined by the presence of circulating antibodies. However, there is little or no information about the frequency of anticardiac antibodies present in failing human myocardium. Methods: Clinical data and myocardial tissue samples were analyzed to determine the role of humoral immunity in patients with chronic heart failure (CHF) in different settings. Results: Anticardiac antibodies were found present in failing hearts but not in normal control hearts. Further, the level of expression of these anticardiac antibodies changed with the severity of the disease state; and in patients with acute heart failure, we found selective activation of B cells. Finally, treatment of CHF patients with therapeutic plasma exchange, a strategy that removes circulating antibodies, resulted in a reduction in anticardiac antibody deposition and improvements in cardiac function. Conclusion: These data collectively suggest a role of humoral immunity in the progression of heart failure. J. Clin. Apheresis, 2010. © 2010 Wiley‐Liss, Inc.     

Erratum to: Enthesitis in seronegative spondyloarthropathies with special attention to the knee joint by MRI: a step forward toward understanding disease pathogenesis

Clinical Rheumatology -