Kinetics and apoptotic profile of circulating endothelial cells as prognostic factors for induction treatment failure in newly diagnosed acute myeloid leukemia patients

The circulating endothelial cells (CEC) are proposed to be a noninvasive marker of angiogenesis. Recent data suggest that endothelial cells may enhance the survival and proliferation of leukemic blasts and mediate chemotherapy resistance in acute myeloid leukemia (AML). We analyzed CEC count by the four-color flow cytometry in AML and healthy subjects. We evaluated the kinetics of mature CEC, both resting (rCEC) and activated (aCEC), as well as progenitor (CEPC) and apoptotic CEC (CEC^AnnV+) in AML patients treated with standard chemotherapy and their influence on response to treatment and overall survival. We found significantly higher numbers of aCEC, rCEC, CEPC, and CEC^AnnV+ in AML patients than in healthy controls. The elevated CEPC and absolute blood counts in peripheral blood as well as the low CEC^AnnV+ number were associated with higher probability of induction treatment failure. aCEC, rCEC, CEPC, and CEC^AnnV+ counts determined in complete remission (CR) were significantly lower than those found at diagnosis. In those CR patients, a significant decrease in the CEC count and increase in the number of CEC^AnnV+ were observed already 24h after the first dose of chemotherapy. In refractory AML, the aCEC, rCEC, CEPC, and CEC^AnnV+ counts assessed before and after induction chemotherapy did not differ significantly, and a significant decrease in CEC count and increase in CEC^AnnV+ number were noted only after the last dose of chemotherapy. The number of CEC is significantly higher in AML patients than in healthy subjects and correlates with response to treatment. The evaluation of CEC kinetics and apoptotic profile may be a promising tool to select AML patients with poor response to chemotherapy who may benefit from antiangiogenic therapies.     

Circulating endothelial cells in patients with acute myeloid leukemia

OBJECTIVES: The circulating endothelial cells (CEC) are proposed to be a non-invasive marker of angiogenesis. The level of CEC in peripheral blood (PB) of acute myeloid leukemia (AML) patients has not been investigated prior to this study. We evaluated the count of resting (rCEC), activated (aCEC) and endothelial progenitor cells (CEPC) in the PB of AML and healthy subjects. In addition we correlated the levels of CEC with disease status, known prognostic factors and response to treatment. METHODS: CEC were quantified by utilizing four-color flow cytometry procedures in 48 AML patients at the time of diagnosis and 29 healthy controls. Additionally, measurements were again taken after the first course of induction treatment in 12 of the patients. RESULTS: The numbers of aCEC, rCEC and CEPC were significantly higher in the AML patients than in the controls (P < 0.0001, P < 0.0001 and P < 0.001, respectively). The CEC count was significantly higher in the AML patients with white blood cell count (WBC) >15 G/L, elevated lactic dehydrogenase (LDH) levels and a higher (over median) absolute blasts count (ABC) in PB than in the group with WBC <15 G/L (P < 0.03), a normal LDH level (P < 0.03) and a lower (相似文献   

Markers of endothelial damage and repair in Takayasu arteritis: Are they associated with disease activity?

The current clinical and laboratory parameters of Takayasu arteritis (TA) are insufficient for proper assessment of disease activity. The aims of this study were to investigate the markers of endothelial injury and repair, including circulating endothelial cell (CEC), circulating endothelial progenitor cell (CEPC) and vascular endothelial growth factor (VEGF), and evaluate their associations with disease activity in patients with TA. Thirty-two patients with TA and 30 healthy age- and sex-matched controls were included in this study. Disease activity was assessed in TA patients using various tools, including Kerr’s criteria, the Indian Takayasu’s Arteritis Scoring (ITAS2010) and physician’s global assessment (PGA). CECs and CEPCs were measured by flow cytometry, and VEGF was measured using an enzyme-linked immunosorbent assay. The CEC level was found to be higher in TA patients than in the healthy controls (HC) (p < 0.001). There was no significant difference in CEC level between the active and inactive patients, but its level was slightly correlated with C-reactive protein (CRP) level. CEPC and VEGF levels in TA patients with active disease were higher than those in the inactive patients and HC. CEPC and VEGF levels were positively correlated with ITAS-CRP and PGA scores. This study shows increased level of CEC in patients with TA. It also suggests that the CEPC and VEGF levels may be correlated with disease activity.     

Effect of smoking cessation on the number and adhesive properties of early outgrowth endothelial progenitor cells

Background

Endothelial progenitor cells participate in angiogenesis and vascular repair, and cardiovascular risk factors may reduce their numbers or impair their functional properties. Cigarette smoking is a leading cause of preventable cardiovascular death, however, the functional properties of these cells before and after discontinuation of tobacco use have not been systematically analyzed.

Methods

We examined changes in the number and function of early outgrowth endothelial progenitor cells (EPC), isolated from individuals (n = 144; mean age, 47.8 ± 12.0 years; 43% males; more than 50% with additional cardiovascular risk factors or disease) who successfully completed a 5-week smoking cessation (SC) programme.

Results

SC significantly reduced total white blood cell count (WBC; P < 0.0001), plasma LDL cholesterol (P = 0.0002) and fibrinogen (P < 0.0001) levels, but did not alter the number of circulating CD34⁺, VEGFR2⁺ or CD34⁺, CD133⁺ cells (P = 0.14 and 0.57, respectively). Fewer acLDL⁺, lectin⁺ cells could be expanded from peripheral blood mononuclear cells in comparison to baseline (P < 0.001). Furthermore, SC was associated with reduced EPC adhesion to fibronectin (P < 0.001) or TNFα-activated endothelial cells (P = 0.003), and a diminished incorporation of EPC into endothelial cell networks (P = 0.035). Mechanistically, significantly reduced β1- and β2-integrin expression (P < 0.001 and 0.007) and lower contents of intracellular reactive oxygen species (P < 0.007) were detected in EPC following SC, in addition to reduced plasma asymmetric dimethyl-L-arginine (ADMA) levels (P = 0.0003).

Conclusions

Our findings suggest that the oxidative and inflammatory stress reduction associated with smoking cessation impair the adhesiveness of monocyte-derived EPC.     

Comparative study on circulating endothelial progenitor cells in systemic lupus erythematosus patients at active stage

Circulating endothelial progenitor cells (CEPCs) play an important role in the process of atherosclerosis. Most previous studies on CEPC in systemic lupus erythematosus (SLE) patients were on their number and some functions and the results were not consistent. No studies on their anti-inflammatory function and integrated status were reported. The purpose of this study was to determine the number, function (including anti-inflammatory function), and the integrated status of CEPCs in active SLE patients. The study was performed in 35 active SLE patients (28 females, 7 males) and 35 age-and gender-matched healthy controls. CEPC number was determined by Fluorescence-Activated Cell Sorting. Proliferation capacity of CEPC was assessed by PCNA staining. Adhesion capacity of CEPC to fibronectin and adhesion capacity of THP1 cell to CEPC were determined by cell adhesion assay. Migratory capacity of CEPC was measured by transwell chamber assay and the potential to form tubes on Matrigel of CEPC was determined by in vivo tube formation on Matrigel test. The expression of inducible nitric oxide synthase (iNOS) and interleukin-6 (IL-6) assessed by quantitative PCR as well as the expression of intercellular adhesion molecule-1 (ICAM-1) and phosphorylated-Akt (p-Akt) assessed by western-blotting were used to evaluate the anti-inflammatory function and cell status of CEPCs. The number of CEPC in SLE patients was not different from that in control (p > 0.05). Proliferation capacity of CEPC was decreased in active SLE patients (p = 0.027). Adhesion capacity of CEPC to fibronectin was decreased (p = 0.04) in SLE patients and adhesion capacity of THP1 cell to CEPC was increased in SLE patients (p < 0.001). Migratory activity was reduced in patient CEPCs (p < 0.001). Capacity of CEPCs to form tube on Matrigel was decreased in SLE patients (p < 0.001). Expression of iNOS and IL-6 (p < 0.001, p = 0.006, respectively) and ICAM-1 were increased in CEPC of SLE patients and expression of p-Akt was decreased in CEPC of SLE patients. Our data show that CEPC number in active SLE patients was not significantly different from healthy controls, but their functions were partly impaired, including proliferation, adhesion, migration, and tube formation. Bad cell status and increased susceptibility to inflammatory process of CEPCs in active SLE were also observed in our study.     

Prognosis of patients with haematological malignancies admitted to the intensive care unit: Sequential Organ Failure Assessment (SOFA) trend is a powerful predictor of mortality

Background

The prognosis of patients with haematological malignancies who are admitted to the ICU is generally poor. In order to optimize care, it is important to be able to determine which patients are most likely to benefit from continuation of treatment after ICU admission.

Methods

Data of 86 patients with a haematological malignancy consecutively admitted to the ICU of Maastricht University Medical Centre were examined in a retrospective cohort study in order to identify clinically useful prognostic parameters.

Results

ICU mortality was 56% and in-hospital mortality was 65%. Non-survivors had higher APACHE-II and SOFA scores compared with survivors (32 ± 8.0 versus 25 ± 6.5 and 11.5 ± 3.1 versus 8.5 ± 3.0, respectively). The mortality rate was significantly higher in patients with an increasing SOFA score (≥ 2 points) compared with patients with an unchanged or decreasing SOFA score (72% versus 58% and 21%, respectively). Mortality was also higher in patients requiring invasive mechanical ventilation or inotropic/vasopressor therapy.

Conclusion

The mortality rate among patients with haematological malignancies who are admitted to the ICU is high and mainly associated with the severity of illness, as reflected by more severe and worsening organ failure and a need for mechanical ventilation or inotropic/vasopressor therapy. Several factors appear to be associated with a poor outcome, but no absolute predictors of mortality could be identified, although the results suggest that changes in the SOFA score during the stay in the ICU can be helpful in the decision making about the continuation or discontinuation of treatment.     

SOFA score in septic patients: incremental prognostic value over age,comorbidities, and parameters of sepsis severity

Several widely used scoring systems for septic patients have been validated in an ICU setting, and may not be appropriate for other settings like Emergency Departments (ED) or High-Dependency Units (HDU), where a relevant number of these patients are managed. The purpose of this study is to find reliable tools for prognostic assessment of septic patients managed in an ED-HDU. In 742 patients diagnosed with sepsis/severe sepsis/septic shock, not-intubated, admitted in ED between June 2008 and April 2016, SOFA, qSOFA, PIRO, MEWS, Charlson Comorbidity Index, MEDS, and APACHE II were calculated at ED admission (T0); SOFA and MEWS were also calculated after 24 h of ED-High-Dependency Unit stay (T1). Discrimination and incremental prognostic value of SOFA score over demographic data and parameters of sepsis severity were tested. Primary outcome is 28-day mortality. Twenty-eight day mortality rate is 31%. The different scores show a modest-to-moderate discrimination (T0 SOFA 0.695; T1 SOFA 0.741; qSOFA 0.625; T0 MEWS 0.662; T1 MEWS 0.729; PIRO: 0.646; APACHE II 0.756; Charlson Comorbidity Index 0.596; MEDS 0.674, all p < 0.001). At a multivariate stepwise Cox analysis, including age, Charlson Comorbidity Index, MEWS, and lactates, SOFA shows an incremental prognostic ability both at T0 (RR 1.165, IC 95% 1.009–1.224, p < 0.0001) and T1 (RR 1.168, IC 95% 1.104–1.234, p < 0.0001). SOFA score shows a moderate prognostic stratification ability, and demonstrates an incremental prognostic value over the previous medical conditions and clinical parameters in septic patients.     

Effectiveness of polymyxin B-direct hemoperfusion (PMX-DHP) therapy using a polymyxin B-immobilized fiber column in patients with post-esophagectomy sepsis

Background

Post-esophagectomy complications have an extremely poor prognosis. Recently, polymyxin B-direct hemoperfusion (PMX-DHP) therapy using a polymyxin B-immobilized fiber column was reported to be beneficial in gram-negative and/or gram-positive bacterial sepsis. The present retrospective study investigated the effectiveness and safety of PMX-DHP therapy in severe sepsis or septic shock after esophagectomy.

Methods

Fifteen severe sepsis or septic shock patients were included. Seven (four, pneumonia; two, anastomotic leakage; and one, reconstructed colon necrosis) patients received 2–5 h of PMX-DHP therapy (PMX-DHP therapy group), whereas 8 (three, pneumonia; three, anastomotic leakage; and two, gastric tube necrosis) received conventional therapy (control group).

Results

Length of stay in the intensive care unit (ICU) was significantly shorter in the PMX-DHP therapy group than in the conventional therapy group (P = 0.040). In the comparison of pre- and post-PMX-DHP therapy groups, the total Sequential Organ Failure Assessment (SOFA) score, respiratory system score, and P/F ratio improved (P = 0.0027, P = 0.025, and P = 0.0087, respectively) in the post-PMX-DHP therapy group. In the comparison of conventional and PMX-DHP therapy groups, the variations in the total SOFA score, respiratory system score, and P/F ratio improved (P = 0.019, P = 0.0063, and P = 0.0015, respectively) in the PMX-DHP therapy group. Moreover, the respiratory system score was lower (P = 0.0062) in the PMX-DHP therapy group at the time of discharge from the ICU. No adverse effects were observed during the course of PMX-DHP therapy.

Conclusions

PMX-DHP therapy was safe and effective in improving respiratory and general conditions of patients with severe sepsis and septic shock after esophagectomy and decreased the length of stay in the ICU.     

Pioglitazone delays proximal tubule dysfunction and improves cerebral vessel endothelial dysfunction in normoalbuminuric people with type 2 diabetes mellitus

Aim

The renal and cerebral protective effects of pioglitazone were assessed in normoalbuminuric patients with type 2 diabetes mellitus (DM).

Methods

A total of 68 normoalbuminuric type 2 DM patients were enrolled in a one-year open-label randomized controlled trial: 34 patients (pioglitazone-metformin) vs. 34 patients (glimepiride-metformin). All patients were assessed concerning urinary albumin: creatinine ratio (UACR), urinary alpha1-microglobulin, urinary beta2-microglobulin, plasma asymmetric dymethyl-arginine (ADMA), GFR, hsC-reactive protein, fibrinogen, HbA1c; pulsatility index, resistance index in the internal carotid artery and middle cerebral artery, intima-media thickness in the common carotid artery; cerebrovascular reactivity was evaluated through the breath-holding test.

Results

At 1 year there were differences between groups regarding ADMA, urinary beta2-microglobulin, urinary alpha1-microglobulin, parameters of inflammation, serum creatinine, GFR, UACR, the cerebral haemodynamic indices. Significant correlations were found between alpha 1-microglobulin-UACR (R² = 0.143; P = 0.001) and GFR (R² = 0.081; P = 0.01); beta2-microglobulin-UACR (R² = 0.241; P = 0.0001) and GFR (R² = 0.064; P = 0.036); ADMA-GFR (R² = 0.338; P = 0.0001), parameters of inflammation, HbA1c, duration of DM, cerebral indices. There were no correlations between ADMA-UACR, urinary alpha1-microglobulin and beta2-microglobulin.

Conclusion

Proximal tubule (PT) dysfunction precedes albuminuria and is dissociated from endothelial dysfunction in patients with type 2 DM. Pioglitazone delays PT dysfunction and improves cerebral vessels endothelial dysfunction in normoalbuminuric patients with type 2 DM. Key words: proximal tubule; endothelial dysfunction, albuminuria; pioglitazone.     

Temporal and venepuncture-related decline in circulating endothelial cell capture from mixed venous blood

Background: The quantification of circulating endothelial cells (CECs) in whole blood has evolved as a novel method for the assessment of endothelial function, although major methodological issues remain. We hypothesized that there is a temporal decline in CEC counts in static venesected blood and that venepuncture itself may lead to increased CEC detachment. Methods: CEC isolation was performed using the immunobead method. For the temporal decline experiment, we included 52 patients presenting with acute coronary syndrome (ACS). We performed CEC counts immediately and at 4 and 24 h later. For the venepuncture decline experiment, we studied 40 patients with stable cardiovascular disease (CVD). CEC counts were determined from the first 4 mL of aspirated venous blood and compared with counts obtained from a subsequent 4 mL sample of blood after at least 7.5 mL of blood had been collected. Results: Among the ACS patients there was a significant temporal decline in CEC counts in static venous blood over a 24 h period (p = 0.013). Among the patients with stable CVD, the median CEC counts obtained from the initial 4 mL of aspirated venous blood were significantly higher (by 32%) than that obtained from the later 4 mL of aspirated venous blood (p = 0.041). Conclusions: We demonstrated a significant temporal fall in CEC numbers in static venous blood over 24 h following venesection. Furthermore, we have shown that CEC counts are higher in the initial aspirated blood compared with that aspirated from the same needle subsequently. These data would have implications for how CEC determination is undertaken by researchers in studies related to ACS or CVD.     

Coronary flow reserve is associated with tissue ischemia and is an additive predictor of intensive care unit mortality to traditional risk scores in septic shock

Background

Reduced coronary velocity flow reserve (CFR) is associated with poor outcome in patients with cardiovascular disease. We investigated whether CFR is associated with tissue ischemia and acidosis, impaired myocardial deformation and adverse outcome in patients with septic shock.

Methods

In 70 mechanically-ventilated patients with septic shock, we examined: a) S′ and E′ mitral annular velocities using tissue Doppler imaging (TDI), b) CFR of the left anterior descending artery after adenosine infusion using transesophageal Doppler echocardiography and c) lactate, pyruvate and glycerol in tissue by means of a microdialysis (MD) catheter inserted into the subcutaneous adipose tissue as markers of tissue ischemia and acidosis. SOFA and APACHE II prognostic scores and mortality in the intensive care unit (ICU) were recorded.

Results

Reduced CFR, S′ and E′ as well as increased E/E′ correlated with increased SOFA, APACHE II and MD lactate to pyruvate ratio (p < 0.05 for all correlations). Impaired TDI markers also correlated with increased MD glycerol (p < 0.05). Reduced CFR correlated with decreased E′ (p < 0.05). CFR was 1.8 ± 0.42 in non-survivors (n = 34) versus 2.08 ± 0.44 in survivors (p = 0.007). A CFR < 1.90 predicted mortality with sensitivity of 70% and specificity of 69% (area under the curve 77%; p = 0.003). CFR had an additive value to APACHE (chi-square change: 4.358, p = 0.03) and SOFA (chi-square change: 3.692, p = 0.04) for the prediction of mortality.

Conclusion

Tissue ischemia and acidosis is a common pathophysiological link between decreased CFR and impaired LV myocardial deformation in septic shock. CFR is an additive predictor of ICU mortality to traditional risk scores in septic shock.     

Effects of continuous positive airway pressure on endothelial function and circulating progenitor cells in obstructive sleep apnoea: A randomised sham-controlled study

Objective

Obstructive sleep apnoea (OSA) is characterised by reoccurring apnoeas and hypopneas, causing repetitive hypoxia and reoxygenation, and is associated with endothelial dysfunction and reduced levels of circulating progenitor cells (CPCs). The potential to improve endothelial function and CPC levels in people with OSA by preventing hypoxic episodes with Continuous Positive Airway Pressure (CPAP) was investigated in a sham-controlled CPAP study.

Methods

Men with moderate-to-severe OSA (mean ± SD: age = 49 ± 12 y, apnoea hypopnea index (AHI) = 37.6 ± 16.4 events/h, body mass index = 31.5 ± 5.7 kg/m²) who were CPAP naïve without diabetes mellitus were randomised in a 12-week double-blind sham-controlled parallel group study to receive either active (n = 25) or sham (n = 21) CPAP. CPCs, isolated from blood, were measured by flow cytometry and by co-staining cultured cells (7 days) with acetylated low-density lipoprotein (acLDL) and lectin. Endothelial function was assessed by peripheral arterial tonometry (PAT).

Results

Compared to sham, CPAP significantly decreased AHI (mean between-group difference − 36.0 events/h; 95%CI, − 49.7 to − 22.3, p < 0.0001) after 12 weeks. Despite this improvement in AHI, CPAP had no effect on change in CPC levels (including CD34⁺/KDR⁺ (565 cells/mL; − 977 to 2106, p = 0.45), CD34⁺/KDR⁺/CD45⁻ (37.0 cells/mL; − 17.7 to 85.7, p = 0.13), acLDL⁺/lectin⁺ (− 43.1 cells/field, − 247 to 161, p = 0.67)) or change in endothelial function (0.27; − 0.14 to 0.67, p = 0.19) compared to sham therapy.

Conclusions

Despite the improvement in OSA parameters and ablation of apnoeic events by CPAP, CPC counts and endothelial function in men with moderate-to-severe OSA were not significantly improved after 12 weeks of therapeutic CPAP when compared to sham control.     

Effects of levothyroxine treatment on insulin sensitivity,endothelial function and risk factors of atherosclerosis in hypothyroid women

Objectives

Contradictory results are encountered in literature regarding the effects of hypothyroidism on the risk factors of atherosclerosis. We aimed to explore the changes in atherosclerotic risk factors and insulin sensitivity before and after levothyroxine replacement therapy in women with primary hypothyroidism and compare with that of healthy controls.

Patients and methods

Twelve patients (mean age of 34 ± 11.7 years) without an evident disease except for primary hypothyroidism (TSH ≥ 20 mIU/L) and eleven euthyroid, age-matched (33.8 ± 8.4 years) female volunteers as controls were included. Baseline thyroid hormones, lipid parameters, homocysteine, fibrinogen levels were measured in both groups. Flow-mediated endothelial-dependent vasodilatation (FMD) method was used to evaluate endothelial dysfunction. Insulin sensitivity was assessed by M values based on euglycemic hyperinsulinemic clamp technique. The same measurements were performed after 6 months of levothyroxine treatment and recovery of euthyroid state in hypothyroid patients.

Results

Treatment reduced total cholesterol (P < 0.005), LDL-cholesterol (P < 0.005), lipoprotein(a) (P < 0.01), fibrinogen (P < 0.0001) and homocysteine (P < 0.0005) levels. Treatment significantly improved M values of hypothyroid patients (3.68 ± 1.53 mg/kg.min vs 6.02 ± 1.21 mg/kg.min, P < 0.0001) and FMD (9.1 ± 3.7% vs 16.4 ± 4.4%, hypothyroid vs euthyroid, P < 0.0001). Significant correlations were found between M values and TSH (r = −0.6, P < 0.005), fibrinogen (r = −0.53, P < 0.01) measurements, free T3 (r = 0.51, P < 0.02) and free T4 (r = 0.49, P < 0.02) levels. FMD was significantly correlated with fibrinogen levels (r = −0.49, P < 0.05).

Conclusion

Insulin resistance, endothelial dysfunction, atherosclerotic risk markers improves with treatment of hypothyroidism.     

Genetic variability on adiponectin gene affects myocardial infarction risk: The role of endothelial dysfunction

Background

Adiponectin is an adipokine with an important role in cardiovascular system conferring anti-inflammatory and anti-atherogenic effects. Two common single nucleotide polymorphisms (SNP) on adiponectin gene, rs2241766 and rs1501299, have been associated with insulin resistance and diabetes mellitus risk however their effects on cardiovascular risk remain unclear. We examined the impact of rs2241766 and rs1501299 on circulating adiponectin levels, endothelial function and cardiovascular disease risk.

Methods

We recruited in total 594 subjects; 462 patients with angiographically confirmed coronary artery disease (CAD) and 132 controls matched for age and gender. rs2241766 and rs1501299 were genotyped by polymerase chain reaction and restriction endonuclease digestion. Serum adiponectin levels were determined by enzyme-linked immunosorbent assay. Endothelial function was assessed by the flow mediated dilatation (FMD) of the brachial artery.

Results

rs2241766 had no effects on circulating adiponectin levels or FMD. In subjects without CAD, carriers of the T/T alleles at rs1501299 had lower adiponectin levels (p = 0.001) and impaired endothelial function (p < 0.05). After multivariate adjustment none of the SNPs had any effect on CAD risk. However, carriers of the T allele at rs1501299 were at increased myocardial infarction (MI) risk, independently of classic risk factors (OR = 2.558 [95%CI = 1.587–4.123], p = 0.0001). The number of T alleles in both SNPs was strongly associated with MI history (p = 0.0001).

Conclusions

rs1501299 polymorphism of adiponectin gene affects circulating adiponectin levels and endothelial function in subjects without CAD. Presence of the T variant at rs1501299 on adiponectin gene is independently associated with increased myocardial infarction risk.     

Anti-endothelial and anti-neuronal effects from auto-antibodies in subsets of adult diabetes having a cluster of microvascular complications

Aims

To test autoantibodies from subsets of diabetes with painful neuropathy, maculopathy and nephropathy for effects in neurons.

Methods

Protein-A eluates from plasma of 27 diabetic and 19 age-matched controls were tested for effects on endothelial cell survival, and neurite outgrowth in rat pheochromocytoma PC12 cells. Painful diabetic neuropathy or control autoantibodies were compared for binding to PC12-derived heparan sulfate proteoglycans. The mechanism of the effects from pathologic autoantibodies was investigated by changes in intracellular calcium in endothelial cells, whole cell current in neurons, or using the Rho kinase inhibitor Y27632.

Results

Autoantibodies from diabetic patients with maculopathy, nephropathy, and painful neuropathy (n = 5) caused significantly greater mean inhibition of neurite outgrowth (p < 0.005) than diabetic or control patients with fewer or no complications (n = 30). Painful diabetic autoantibodies (3 μg/mL) bound neuronal heparan sulfate proteoglycan (HSPG) more than autoantibodies from diabetic or control subjects without painful neuropathy (p < .0001). Inhibition of PC12 neurite outgrowth by the painful neuropathy antibodies was completely prevented by 1 μM concentrations of Y27632.

Conclusion

These results suggest anti-endothelial and anti-neuronal effects from auto-antibodies in a subset of diabetic patients with a cluster of microvascular complications.     

Risk factors for 28-day mortality in elderly patients with extended-spectrum β-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae bacteremia

Gram-negative bacteremia is common in elderly patients and, compared with younger patients, mortality rates in bacteremic elderly patients are high. ESBL-producing organisms were one of the most important risk factors associated with mortality. In addition, older age is one of risk factors for colonization or infection with ESBL-producing organisms. We conducted a retrospective cohort study to evaluate risk factors of all-cause 28-day mortality in elderly patients with ESBL-producing Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) bacteremia. Patients aged 65 years or older, who had one or more blood cultures positive for E. coli and K. pneumoniae and who were hospitalized between January 2006 and December 2010 at a tertiary-care teaching hospital, were included. 191 bacteremic elderly patients were eligible for the study. The all-cause 28-day mortality rate was 24.6% (47/191). In multivariate analysis, prior antimicrobial therapy (p = 0.014) and an elevated SOFA score (p < 0.001) were independent risk factors for increased mortality, while urinary tract infection (UTI) was an independent determinant for non-mortality (p = 0.011). In the current study, prior antimicrobial therapy within 30 days, an elevated SOFA score and nonurinary source of infection were significantly associated with adverse outcomes in elderly patients with ESBL-producing gram-negative bacteremia.     

Expression of miR-126 and miR-508-5p in endothelial progenitor cells is associated with the prognosis of chronic heart failure patients

Background

MicroRNA (miRNA) expression profiles in endothelial progenitor cells (EPCs) contribute to EPC dysfunction in patients suffering from coronary artery disease. However, it remains unclear whether miRNA expression in EPCs is associated with the prognosis of chronic heart failure (CHF) secondary to ischemic cardiomyopathy (ICM) or non-ischemic cardiomyopathy (NICM).

Methods and results

One hundred six patients with CHF (55 ICM and 51 NICM) and 30 healthy controls were followed until the end of 24 months or when the end point was obtained (cardiovascular death). The miRNA expression profile was analyzed by TaqMan Human MicroRNA Array Set v2.0 in 30 randomly assigned samples (ICM = 10, NICM = 10, and healthy controls = 10). During the 24-month follow-up, 26 patients died from cardiovascular disease. Sixteen miRNAs (miR-126, miR-508-5p, miR-34a, miR-210, miR-490-3p, miR-513-5p, miR-517c, miR-518e, miR-589, miR-220c, miR-200a*, miR-186*, miR-7i*, miR-200b*, miR-595, and miR-662) were found to be differentially expressed between ICM and NICM patients. Survival analysis showed that miR-126 and miR-508-5p levels in EPCs were independent prognostic factors (P = 0.003; HR (hazard ratio): 0.19; 95% CI (confidence intervals): 0.06–0.58, P = 0.002; HR: 2.292; 95% CI: 1.37–3.84) for the outcome of ICM or NICM patients with CHF. Pathway enrichment analysis showed that the angiogenesis pathway was the most likely pathway regulated by miR-126 and miR-508-5p.

Conclusions

The miRNAs miR-126 and miR-508-5p are associated with the outcome of ICM and NICM patients with CHF. These two miRNAs could be useful in the diagnosis of CHF patients, and might provide novel targets for prevention and treatment of CHF.     

The kinetics and apoptotic profile of circulating endothelial cells in autologous hematopoietic stem cell transplantation in patients with lymphoproliferative disorders

In neoplastic disorders, endothelial cells take part in tumor progression and also influence the recovery of hematopoiesis after high-dose chemotherapy. Measurements of circulating endothelial cells (CEC), their subsets and kinetics were taken in patients with lymphoid malignancies (37 multiple myeloma, ten lymphoma) during autologous hematopoietic stem cell transplantation (HSCT). CEC were evaluated by four-color flow cytometry at different time points. Additionally levels of angiopoietins 1 and 2 were evaluated by ELISA assay. The baseline number of CECs and their subsets in patients were higher than in the control group. The median CEC number dropped significantly after transplantation (from 9.5/μL to 6.2/μL, p?<?0.001). Apoptosis of CECs 24 h after chemotherapy was enhanced in comparison to baseline values (median apoptotic CEC number 4.15/μL vs 3.1/μL; p?<?0,001). The time for neutrophil engraftment was shorter for patients with a low apoptotic CEC count at baseline as compared to those with a high apoptotic CEC count (median time to engraftment 13 vs. 16 days respectively, p?=?0.04). We observed an adverse correlation of progenitor CEC numbers measured 1 h after transplantation with the time to neutrophil engraftment (r?=??0.49, p?=?0.008). We also found a negative correlation between the number of CECs originating from microvessels measured 1 h after transplantation, and the time to neutrophil engraftment (r?=??0.39, p?=?0.04). Baseline angiopoietins 1 and 2 concentration did not influence the post-transplant regeneration time. CEC numbers significantly change during autologous HSCT. Our results suggest that progenitor CECs and CECs derived from microvessels both take part in successful engraftment.     

Insulin resistance and endothelial function in children and adolescents

Aims

Insulin resistance (IR) impairs cellular response to insulin due to a dysfunction in glucose metabolism, associated with an increased cardiovascular risk. The aim of our study was to investigate the relationship among homeostasis model assessment index (HOMA index), endothelial function and vascular morphology in order to better stratify cardiovascular risk in children and adolescents.

Methods

A total of 150 children and adolescents (55 pre-pubertal, mean age 10.4 ± 3.1 years) were enrolled. Anthropometric [body mass index (BMI), waist circumference (WC)], laboratory [blood lipids, inflammatory markers, insulinemia, glycemia], HOMA index and ultrasound parameters [flow-mediated dilatation (FMD), common carotid intima–media thickness (cIMT) and antero-posterior diameter of infra-renal abdominal aorta (APAO)] were assessed.

Results

cIMT was positively related to age (r = 0.274, p < 0.01), BMI (r = 0.318, p < 0.01), WC (r = 0.315, p < 0.01) and triglycerides (r = 0.230, p < 0.01). APAO measurements showed a linear positive correlation with age (r = 0.435, p < 0.01), BMI (r = 0.505, p < 0.01), WC (r = 0.487, p < 0.01), triglycerides (r = 0.280, p < 0.01), C-reactive protein (r = 0.209, p < 0.05), fasting insulin (r = 0.378, p < 0.01) and HOMA index (r = 0.345, p < 0.01). FMD was inversely related to age (r = − 0.251, p < 0.01), rough BMI (r = − 0.318, p < 0.01), WC (r = − 0.340, p < 0.01), fasting insulin (r = − 0.281, p < 0.01) and HOMA index (r = − 0.282, p < 0.01). Multiple regression analysis found no influence of HOMA index on APAO and cIMT. HOMA index was an independent predictor for brachial artery FMD worsening after the statistical adjustment.

Conclusion

HOMA index increase induced a worsening in endothelial function since childhood.     

Circulating endothelial cells in patients with septic shock

The vascular endothelium has a central role in the control of microvascular tone, and it has been proposed that vascular endothelial damage occurs in septic shock, producing multiorgan failure. We have developed a method of detecting circulating endothelial cells (EC) that provides direct evidence of EC shedding in human sepsis. Human umbilical vein endothelial cells (HUVEC) were seeded in whole blood and recovered by isopycnic centrifugation to validate the technique. Blood samples were subsequently taken from 11 healthy volunteers, nine ventilated intensive care unit (ICU) control patients without sepsis, eight patients with sepsis but without shock, and 15 patients with septic shock. EC were identified by indirect immunofluorescence, using antibodies to von Willebrand factor (vWf) and the vascular endothelial growth factor receptor KDR. Mean HUVEC recovery was 86% for 20 to 100 seeded cells/ml of blood. vWf-positive EC counts per milliliter were significantly higher (analysis of variance [ANOVA], p < 0.0001) in patients with sepsis (16.1 +/- 2.7 [mean +/- SEM]) and septic shock (30.1 +/- 3.3) than in healthy (1.9 +/- 0.5) or ICU controls (2.6 +/- 0.6). KDR-positive EC counts per milliliter were also significantly higher (ANOVA, p < 0.0001) in patients with sepsis (4.2 +/- 1.1/ml) and septic shock (10.4 +/- 1.2/ml) than in healthy (0.7 +/- 0.3/ml) or ICU controls (0.5 +/- 0.2/ml). Cell counts made with anti-vWf antibody were consistently higher than those made with anti KDR antibody, but correlation between the two counts was high (r(2) = 0.93). The number of circulating KDR-positive EC was significantly higher in patients who died of septic shock than in survivors (12.0 +/- 1.6/ml versus 7.1 +/- 1.2/ml, p = 0.026). An increase in circulating EC can be identified during sepsis and septic shock. This supports the hypothesis that endothelial damage occurs in human sepsis.