Evidence for non‐functional Dickkopf‐1 (DKK‐1) signaling in chronic lymphocytic leukemia (CLL)

Purpose: Wnt signaling was demonstrated to be activated in chronic lymphocytic leukemia (CLL). It is thought to be responsible for the extended survival of CLL cells in vivo. Dickkopf1 (DKK1) is known to antagonize Wnt signaling by direct high‐affinity binding to the extracellular domain of WNT coreceptor lipoprotein receptor‐related protein 6 (LRP6). The purpose of this study was to investigate the effect of DKK1 in B‐CLL cells in vitro. Methods: Expression of DKK1 was estimated by Western blot and real‐time PCR. B cells from patients with CLL and healthy donors were incubated with recombinant DKK1. Survival was measured by flow cytometry. Primers for real‐time PCR were designed for extracellular domain of LRP6, responsible for DKK1 binding, and the intracellular region, essential for inhibiting GSK3 β. Results: Healthy and CLL cells express equivalent mRNA levels of DKK1 and LRP6. After treatment of CLL cells with recombinant DKK1 (1 μg/mL) for 3 h, there was no change in the levels of phosphorylated β‐catenin and total β‐catenin. Healthy B cells proved to have significantly higher levels of extracellular, DKK1 binding domain of LRP6. We estimated that in CLL cells every 6th LRP6 receptor is lacking the extracellular domain. Discussion: For the first time we show the expression of DKK1 in CLL cells. Unlike in similar tumors, the addition of DKK1 to culture of CLL cells does not inactivate WNT pathway. The reason for this could be the absence of the binding domain of LRP6. On the other hand, a truncated LRP6 without extracellular DKK1 binding domain could lead to an uncontrollable activation of WNT signaling.     

All‐oral therapy with nucleotide inhibitors sofosbuvir and GS‐0938 for 14 days in treatment‐naive genotype 1 hepatitis C (NUCLEAR)

Sofosbuvir and GS‐0938 are distinct nucleotide analogues with activity against hepatitis C virus (HCV) in vitro. We evaluated the antiviral activity and safety of sofosbuvir and GS‐0938 alone and in combination in HCV genotype 1 patients. In this double‐blind study, 40 treatment‐naïve patients were randomly assigned to 4 treatment cohorts: (i) GS‐0938 for 14 days, (ii) GS‐0938 for 7 days followed by GS‐0938 plus sofosbuvir for 7 days, (iii) sofosbuvir for 7 days followed by GS‐0938 plus sofosbuvir for 7 days and (iv) GS‐0938 plus sofosbuvir for 14 days. In each arm, 8 patients received active drug and 2 placebo. After 7 days of dosing, patients in all 4 dose groups experienced substantial reductions in HCV RNA, with median declines (Q1, Q3) of ?4.50 (?4.66, ?4.24) in Cohort 1, ?4.55 (?4.97, ?4.13) in Cohort 2, ?4.65 (?4.78, ?4.17) in Cohort 3 and ?4.43 (?4.81, ?4.13) in Cohort 4; patients receiving placebo had essentially no change in HCV RNA (+0.07 log₁₀ IU/mL). Seven days after the end of treatment, the proportions of patients with HCV RNA <15 IU/mL were 4 (50%), 8 (100%), 7 (88%) and 5 (63%) for Cohorts 1–4, respectively, vs 0 for placebo. No viral breakthrough or resistance mutations were observed. No serious adverse events or Grade 3 or 4 adverse events were reported. Sofosbuvir and GS‐0938—alone and in combination—were well tolerated and led to substantial reductions in viral load. Sofosbuvir is undergoing further investigation as a possible backbone of an all‐oral regimen for chronic HCV.     

Cytotoxicity of Acetaldehyde‐Derived Advanced Glycation End‐Products (AA‐AGE) in Alcoholic‐Induced Neuronal Degeneration

Abstract : Background: The Maillard reaction that leads to the formation of advanced glycation end‐products (AGEs) plays an important role in the pathogenesis of angiopathy in diabetic patients, in aging and in neurodegenerative processes. We hypothesize that acetaldehyde (AA), one of the main metabolites of alcohol, may be involved in alcohol‐induced neurotoxicity in vivo by formation of AA‐derived AGEs (AA‐AGE) with brain proteins. Methods: AA‐AGE‐bovine serum albumin (BSA) and AA‐AGE‐rabbit serum albumin (RSA) were prepared as described previously. Antibody specific for AA‐AGE was isolated from rabbit antiserum by affinity chromatography. Primary cortical neuronal cell cultures were prepared as described previously. Results: Incubation of cortical neurons with AA‐AGE produced a dose‐dependent increase in neuronal cell‐death, and the neurotoxicity of AA‐AGE was neutralized by the addition of an anti‐AA‐AGE specific antibody, but not by anti‐N‐ethyllysine (NEL) antibody. The AA‐AGE epitope was detected in human brain of alcoholism. Conclusions: We propose that the structural epitope AA‐AGE is an important toxic moiety for neuronal cells in alcoholism.     

Estimation of monocyte‐chemoattractantprotein‐1 (Mcp‐1) level in patients with lupus nephritis

Aim: To evaluate the use of non‐invasive estimation of CP‐1 in urine as a good indicator for lupus nephritis activity. Methods: The study was conducted on 30 patients with systemic lupus erythematosus (SLE) (group I): 15 of these patients were selected without renal involvement (group I [A]), and the other 15 were selected with evidence of renal involvement (group I [B]). Further 10 age‐ and sex‐matched healthy subjects were taken as a control group (group II). The SLE disease activity index (SLEDAI) was applied. Laboratory investigations done for the studied group of patients included: renal function tests (antinuclear antibody) titer, (anti‐double‐stranded DNA) titer, and monocyte chemotactic protein 1 (MCP‐1) level in serum and urine samples. Results: Serum MCP‐1 was significantly higher in SLE patients with nephritis than in the control group, while no significant difference was found between SLE patients without nephritis and the control group. Urinary MCP‐1 in patients with active lupus nephritis (LN) were significantly higher than both patients with inactive LN and control the group. Urinary MCP‐1 in SLE patients with nephritis was significantly higher than both group I (A) and group II. Urinary MCP‐1 correlated positively with proteinuria, and negatively with creatinine clearance and hemoglobin; thus, urinary MCP‐1 correlates with the severity of nephritis. Conclusion: Urinary and not serum MCP‐1 is a useful invasive technique for the assessment of renal disease activity in patients with LN.     

Structure‐oriented review of 14‐3‐3 protein isoforms in geriatric neuroscience

This review focuses on the possible relevance of 14‐3‐3 proteins in geriatric neuroscience. 14‐3‐3 proteins are mainly localized in the synapses and neuronal cytoplasm. These proteins regulate intracellular signal cascades for differentiation, development, growth, apoptosis and survival. Seven isoforms have so far been identified in mammals. The binding motifs and potential functions of 14‐3‐3 proteins are now recognized to have a wide range of functional relevance. First, we provide a brief summary of the molecular structure and multiple functions of 14‐3‐3 proteins. Second, we review the involvement of 14‐3‐3 proteins in common diseases of geriatric neurology, such as Alzheimer's disease and tauopathies, Parkinson's disease and α‐synucleinopathies, Huntington's disease and polyglutamine diseases, Creutzfeldt–Jakob disease and prion diseases, cerebral infarction, and atherosclerosis. Finally, we discuss the immunohistochemical localization of 14‐3‐3 proteins and its isoforms during the postnatal development of rat brains as a basis for understanding adult neurogenesis. The elucidation of the isoform‐dependent functions of 14‐3‐3 proteins with regard to brain development might be promising for the future development of novel therapeutic interventions for common diseases of geriatric neurology. Geriatr Gerontol Int 2012; ??: ??–??.     

Single‐operator double‐balloon endoscopy (DBE) is as effective as dual‐operator DBE

Background and Study Aims: Double‐balloon endoscopy (DBE) is a new device that allows diagnosis and treatment throughout the entire small intestine. Although the originally described method requires two operators, we have recently developed a method to perform DBE by a single operator. We here assessed the clinical usefulness of this one‐person method in comparison to the conventional two‐person DBE. Patients and Methods: One hundred sixty‐two patients (102 men and 60 women, mean age 59 years) underwent 303 DBE procedures. Total observation time, completion rate of total intestinal and colonic observation, lesion‐discovery rate, and complication rate were retrospectively compared between the one‐person method and the conventional two‐person method of DBE. The one‐person method consists of the Grip and Pinch technique and Keep (or Hold) and Slide technique. Results: The total observation times were 95.5 ± 35.1 min and 96.7 ± 47.5 min by one‐person and two‐person antegrade DBE, respectively, and 103 ± 29.8 min and 111 ± 30.1 min by one‐person and two‐person retrograde DBE, respectively. The completion rate for examination of the entire small intestine was 74.2% in one‐person DBE and 76.5% in two‐person DBE, respectively. The lesion‐discovery rate was 69.0% in one‐person DBE and 65.5% in two‐person DBE, respectively. No significant differences between two methods were found in all measures. Also, no difference was observed in complication rate of the two methods. Conclusions: The single‐operator method for DBE was as efficient as the dual‐operator DBE without any higher risk of complications and, therefore, could replace the conventional dual‐operator method in the future.     

C‐Reactive Protein (CRP)‐Lowering Agents

C‐reactive protein (CRP) plays a role in the pathogenesis of cardiovascular disease. It is a marker and predictor of cardiovascular disease. CRP possesses numerous cardiovascular effects (clotting, generation of oxygen radicals, increase in the expression of adhesion molecules and plasminogen activator inhibitor‐1, plaque destabilization) that could result in cardiovascular disease. This review describes the effects of various cardiovascular drugs on the levels of CRP in health and disease. Cyclooxygenase inhibitors (aspirin, rofecoxib, celecoxib), platelet aggregation inhibitors (clopidogrel, abciximab), lipid lowering agents (statins, ezetimibe, fenofibrate, niacin, diets), β‐adrenoreceptor antagonists and antioxidants (vitamin E), as well as angiotensin converting enzyme (ACE) inhibitors (ramipril, captopril, fosinopril), reduce serum levels of CRP; while enalapril and trandolapril have not been shown to have the same effect. Angiotensin receptor blockers (ARBs) (valsartan, irbesartan, olmesartan, telmisartan) markedly reduce serum levels of CRP. The findings with other ARBs (losartan and candesartan) were inconsistent. Antidiabetic agents (rosiglitazone and pioglitazone) reduce CRP levels, while insulin is ineffective. Calcium channel antagonists have variable effects on CRP levels. Hydrochlorothiazide and oral estrogen do not affect CRP. The CRP‐lowering effect of statins is more pronounced than their lipid lowering effect and is not dependent on their hypolipemic activity. The effect of atorvastatin on CRP seems to be dose‐dependent. CRP‐lowering effect of statins is likely to contribute to the favorable outcome of statin therapy. The data suggest that lipid lowering agents, ACE inhibitors, ARBs, antidiabetic agents, antiinflammatory and antiplatelet agents, vitamin E, and β‐adrenoreceptor antagonists lower serum or plasma levels of CRP, while vitamin C, oral estrogen and hydrochlorothiazide do not affect CRP levels.     

Japan Clinical Oncology Group (JCOG) prognostic index and characterization of long‐term survivors of aggressive adult T‐cell leukaemia‐lymphoma (JCOG0902A)

This study evaluated the clinical features of 276 patients with aggressive adult T‐cell leukaemia‐lymphoma (ATL) in 3 Japan Clinical Oncology Group (JCOG) trials. We assessed the long‐term survivors who survived >5 years and constructed a prognostic index (PI), named the JCOG‐PI, based on covariates obtained by Cox regression analysis. The median survival time (MST) of the entire cohort was 11 months. In 37 patients who survived >5 years, no disease‐related deaths in 10 patients with lymphoma‐type were observed in contrast to the 10 ATL‐related deaths in other types. In multivariate analysis of 193 patients, the JCOG‐PI based on corrected calcium levels and performance status identified moderate and high risk groups with an MST of 14 and 8 months respectively (hazard ratio, 1·926). The JCOG‐PI was reproducible in an external validation. Patients with lymphoma‐type who survived >5 years might have been cured. The JCOG‐PI is valuable for identifying patients with extremely poor prognosis and will be useful for the design of future trials combining new drugs or investigational treatment strategies.     

Risk of dipeptidyl peptidase‐4 (DPP‐4) inhibitors on site‐specific cancer: A systematic review and meta‐analysis

The long‐term impact of dipeptidyl peptidase‐4 (DPP‐4) inhibition is unknown, and there are concerns about the influence of DPP‐4 inhibition on carcinogenesis of the pancreas and thyroid. As DPP‐4 is a rather unselective enzyme present in many tissues, we focused on all specific cancer types. PubMed and EMBASE were searched between January 2005 and April 2017 to identify studies comparing DPP‐4 inhibitors with either placebo or active drugs on cancer risk. Studies were included if they reported on at least one specific cancer outcome and had a follow‐up of at least 1 year after start of drug use. Methodological quality of the studies was assessed by the Cochrane Collaboration's tool and the Newcastle‐Ottawa Scale. Twenty‐five studies met the inclusion criteria (12 randomized controlled trials and 13 observational studies). Sample sizes of the DPP‐4 inhibitor groups ranged from 29 to 8212 patients for randomized controlled trials and from 2422 to 71 137 patients for observational studies. Mean age ranged from 51 to 76 years, and mean follow‐up was 1.5 years. None of the pooled (sensitivity) analyses, except the observational studies studying breast cancer (hazard ratio [95% CI]: 0.76 [0.60‐0.96]), showed evidence for an association between DPP‐4 inhibitors and site‐specific cancer. Also for pancreatic and thyroid cancer, no statistically significant risk was found. Based on the current literature, it is not possible to conclude whether DPP‐4 inhibitors were associated with an increased risk of site‐specific cancer. Future studies should address the methodological limitations and follow patients for a longer period to determine the long‐term cancer risk of DPP‐4 inhibitors.     

Translocations t(X;14)(q28;q11) and t(Y;14)(q12;q11) in T-cell prolymphocytic leukemia

We report a case of T-cell prolymphocytic leukemia (T-PLL) in a 41-year-old male. Classical cytogenetic, spectral karyotyping (SKY) and fluorescence in situ hybridization (FISH) studies of a blood sample obtained at diagnosis revealed the co-existence of t(X;14)(q28;q11), t(Y;14)(q12;q11) and a ring chromosome derived from i(8)(q10). Immunophenotypic studies revealed involvement of T-cell lineage, with proliferation of CD4(-) CD8+. The co-existence of two translocations involving both sex chromosomes in a case of T-PLL is rare. Chromosomal instability associated with the disease progression may have allowed the emergence of cell clones with translocations involving the sex chromosomes and the ring chromosome observed.     

Carbohydrate‐deficient isoforms of transferrin (%CDT) and sialic acid (SA) in iron‐deficiency anemia

This study has investigated the serum levels of carbohydrate‐deficient isoforms of transferrin (CDT) and sialic acid (SA) in iron‐deficiency anemia (IDA). Blood samples were collected from 60 women with IDA and from 20 healthy controls. CDT was estimated by anion‐exchange chromatography on minicolumns followed by photometric detection of transferrin and was expressed as a percentage of total transferrin (%CDT). SA was measured by an enzymatic method. There was no difference in the mean level of %CDT between patients with IDA (2.26%) and control patients (2.05%). SA increased significantly from control level 0.61 to 0.69 g/l in anemic patients. We concluded that elevated concentration of total transferrin in IDA did not change the relative value of low sialylated isoforms (%CDT) and the increase of total SA level in the sera of anemic patients is not related to the increase of total transferrin.     

Ten‐Eleven‐Translocation 2 (TET2) is downregulated in myelodysplastic syndromes

TET2, a member of the ten‐eleven‐translocation (TET) family genes that modify DNA by converting 5‐methylcytosine (5‐mC) to 5‐hydroxymethylcytosine (5‐hmC), is located in chromosome 4q24 and is frequently mutated in myeloid malignancies. The impact of TET2 mutation on survival outcomes is still controversial; however, functional studies have proved that it is a loss‐of‐function mutation that impairs myeloid cell differentiation and contributes to the phenotype of myeloid neoplasia. We, herein, aimed to investigate TET2 expression in patients with myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML). A significantly decreased TET2 expression was observed in bone marrow cells from AML (n = 53) and patients with MDS (n = 64), compared to normal donors (n = 22). In MDS, TET2 expression was significantly reduced in RAEB‐1/RAEB‐2 compared to other WHO 2008 classifications, and a lower TET2 expression was observed at the time of MDS disease progression in four of five patients. In multivariate analysis, low TET2 expression (P = 0.03), male gender (P = 0.02), and WHO 2008 classification (P < 0.0001) were independent predictors of poorer overall survival. These results suggest that defective TET2 expression plays a role in the MDS pathophysiology and predicts survival outcomes in this disease.