Paraoxonase activity in athletes with depleted iron stores and iron-deficient erythropoiesis

ObjectiveThe aim of this study was to evaluate how conditions that precede anaemia (iron store depletion and iron-deficient erythropoiesis) affect human serum paraoxonase PON1 activity.Design and methodsBased on haemoglobin, transferrin saturation and serum ferritin values 119 athletes were divided into three groups: with iron depletion, with deficient erythropoiesis and controls. The following parameters were measured: paraoxonase activity towards paraoxon (POase) and diazoxon (DZOase), lipid hydroperoxides (LOOH), the pro-oxidant-antioxidant balance (PAB), red blood cells (RBC) and lipid status.ResultsSignificant differences were found between athletes with different stages of iron deficiency and controls with respect to PON 1 activity and oxidative stress status parameters (Wilks' Lambda = 0.712, F = 5.241, p < 0.001, η² = 0.156). There was no significant difference between the PON1 192 Q and R polymorphism distribution in the two groups of athletes with different stages of iron deficiency and controls (χ² = 1.086; p = 0.896). PON1 activity was positively correlated with RBCs, haemoglobin, transferrin saturation (p < 0.001) and ferritin (p = 0.037) and negatively correlated with LOOH (p = 0.044) in all three study groups.ConclusionsDeficient erythropoiesis in athletes contributes to impaired PON1 activity. In contrast, iron depletion, regardless of increased oxidative stress, does not affect PON1 activity.     

Significantly elevated C-reactive protein serum levels are associated with very high 30-day mortality rates in hospitalized medical patients

Background:There is insufficient data regarding the differential diagnosis and the prognostic value of significantly elevated serum levels of C-reactive protein (CRP) in hospitalized medical patients.Design and methods:A retrospective review of medical charts of patients admitted to a tertiary hospital's Internal Medicine ward during a period of 1 year who had at least one CRP serum level measurement of 200 mg/L or more.Results:Overall, 341 patients with a mean age of 69.8 ± 1.0 years were included in the study. Acute infection was the most prevalent diagnosis (n = 293; 85.9%) with community-acquired pneumonia being the most common acute infection (n = 115; 33.7%). Non-infectious conditions accounted for 9.1% (n = 31) of the diagnoses and included mainly malignant metastatic diseases (n = 19; 5.6%). Overall, 70 (20.5%) patients died within 30 days of admission. Age and active malignancy, with metastasis or without metastasis, were independently associated with 30-day mortality.Conclusion:Significantly elevated CRP serum levels are associated with bacterial infections, malignant diseases, and very high rates of 30-day mortality in hospitalized medical patients.     

Process evaluation of physical activity counselling with and without the use of mobile technology: A mixed methods study

BackgroundA monitoring-and-feedback tool was developed to stimulate physical activity by giving feedback on physical activity performance to patients and practice nurses. The tool consists of an activity monitor (accelerometer), wirelessly connected to a Smartphone and a web application. Use of this tool is combined with a behaviour change counselling protocol (the Self-management Support Programme) based on the Five A's model (Assess–Advise–Agree–Assist–Arrange).ObjectivesTo examine the reach, implementation and satisfaction with the counselling protocol and the tool.DesignA process evaluation was conducted in two intervention groups of a three-armed cluster randomised controlled trial, in which the counselling protocol was evaluated with (group 1, n = 65) and without (group 2, n = 66) the use of the tool using a mixed methods design.SettingsSixteen family practices in the South of the Netherlands.ParticipantsPractice nurses (n = 20) and their associated physically inactive patients (n = 131), diagnosed with Chronic Obstructive Pulmonary Disease or Type 2 Diabetes, aged between 40 and 70 years old, and having access to a computer with an Internet connection.MethodsSemi structured interviews about the receipt of the intervention were conducted with the nurses and log files were kept regarding the consultations. After the intervention, questionnaires were presented to patients and nurses regarding compliance to and satisfaction with the interventions. Functioning and use of the tool were also evaluated by system and helpdesk logging.ResultsEighty-six percent of patients (group 1: n = 57 and group 2: n = 56) and 90% of nurses (group 1: n = 10 and group 2: n = 9) responded to the questionnaires. The execution of the Self-management Support Programme was adequate; in 83% (group 1: n = 52, group 2: n = 57) of the patients, the number and planning of the consultations were carried out as intended. Eighty-eight percent (n = 50) of the patients in group 1 used the tool until the end of the intervention period. Technical problems occurred in 58% (n = 33). Participants from group 1 were significantly more positive: patients: χ²(2, N = 113) = 11.17, p = 0.004, and nurses: χ²(2, N = 19) = 6.37, p = 0.040. Use of the tool led to greater awareness of the importance of physical activity, more discipline in carrying it out and more enjoyment.ConclusionsThe interventions were adequately executed and received as planned. Patients from both groups appreciated the focus on physical activity and personal attention given by the nurse. The most appreciated aspect of the combined intervention was the tool, although technical problems frequently occurred. Patients with the tool estimated more improvement of physical activity than patients without the tool.     

High density lipoprotein-anionic peptide factor effect on reverse cholesterol transport in type 2 diabetic patients with and without coronary artery disease

ObjectivesTo verify if HDL3 Anionic Peptide Factor (HDL3-APF) is as an apolipoprotein that promotes the reverse cholesterol transport.Design and methodsWe investigated a possible association between plasma HDL3-APF concentration, cholesterol efflux from Fu5AH cells and cholesteryl ester transfer protein (CETP) activity in type 2 diabetic patients with coronary artery disease (CAD) (n = 36), those without CAD (n = 20), and 37 healthy subjects.ResultsPlasma APF concentrations were decreased in diabetics with CAD compared to controls (p < 0.01). Cellular cholesterol efflux was decreased in diabetics without and with CAD, (p < 0.01 and p < 0.001 respectively). CETP activity was significantly elevated in all patient groups. Multiple linear regression analysis shows that cholesterol efflux was independently and positively related only to APF concentrations in controls.ConclusionsAPF is likely to be a key independent factor for promoting cellular cholesterol efflux in healthy subjects. However this association is altered in type 2 diabetes.     

Effect of valproic acid treatment on copper availability in adult epileptic patients

ObjectivesIn rats a significant increase of the copper biliary excretion is produced by valproic acid administration, however, a conclusive study on the possible appearance of copper deficiency in humans during treatment with this drug has still not been carried out.Design and methodsIn 101 adult epileptic patients treated in monotherapy (n = 75) and polytherapy (n = 26) with valproic acid, and 50 healthy controls, were determined serum copper, immunoreactive ceruloplasmin and its oxidase activity against o-dianisidine, in order to calculate the specific oxidase activity (activity per unit mass of enzyme protein) and copper/ceruloplasmin ratio.ResultsSpecific oxidase activity of ceruloplasmin and copper/ceruloplasmin ratio were significantly lower in the groups of patients treated with valproic acid than in the controls. Significant correlations were obtained between both biochemical variables (p < 0001). In 38% of the patients treated in mono or polytherapy, the specific oxidase activities of ceruloplasmin were smaller than the estimated lower limit of reference.ConclusionsThese results, possible due to a diminished copper content of the circulating ceruloplasmin, suggest that marginal or moderate copper deficiency may have a substantial prevalence among patients treated with valproic acid.     

CYP11B2 gene haplotypes independently and in concurrence with aldosterone and aldosterone to renin ratio increase the risk of hypertension

ObjectivesAldosterone synthase produces aldosterone, which regulates electrolytes and thereby blood pressure. Polymorphisms in aldosterone-synthase gene (CYP11B2) may associate with heterogeneous aldosterone production and hypertension. Hence, we investigated ? 344T/C, Iw/Ic polymorphisms of CYP11B2, plasma renin activity (PRA) and aldosterone concentration (PAC).Design and methodsConsecutive ethnically-matched 450 hypertensive patients and 360 controls were screened by PCR-RFLP for genotypes and haplotypes; PRA and PAC were measured.ResultsThe Iw/Ic polymorphism distribution differed significantly between the two groups (LRT χ² = 15.8, df = 2, P = 0.000). The mutant allele-Ic and genotype-Ic/Ic were overrepresented in patients (35% versus 27% and 13% versus 7%). Overrepresentation of T-Ic haplotype in patients was identified as risk haplotype (P = 0.000). Patients had significantly higher PAC and aldosterone-to-renin ratio (ARR; P  = 0.000), which was Ic-allele dependent.ConclusionsThe haplotype T-Ic associated with hypertension susceptibility. Correlation between Ic-allele and raised ARR likely serve in hypertension management.     

Enhanced cortical reperfusion protects coagulation factor XII-deficient mice from ischemic stroke as revealed by high-field MRI

Intrinsic coagulation factor XII deficient (FXII^?/?) mice are protected from ischemic stroke. To elucidate underlying mechanisms we investigated the early ischemic period in vivo by multimodal magnetic resonance imaging (MRI) at 17.6 Tesla.Cerebral ischemia was induced by either transient (60 min) or permanent occlusion of the middle cerebral artery (t/pMCAO). 10 FXII^?/? mice underwent t- , 10 FXII^?/? mice p- and 10 Wildtype (Wt) mice tMCAO. Cerebral blood flow (CBF), diffusion-weighted-imaging (DWI) and T2-relaxometry were measured at 2 h and 24 h after MCAO. Outcome measures were evaluated after motion correction and normalization to atlas space. 2 h after tMCAO CBF reduction was similar in FXII^?/? and Wt mice extending over cortical (CBF (ml/100 g/min) 33.6 ± 6.9 vs. 35.3 ± 4.6, p = 0.42) and subcortical regions (25.7 ± 4.5 vs. 31.6 ± 4.0, p = 0.17). At 24 h, recovery of cortical CBF by +36% was observed only in tMCAO FXII^?/? mice contrasting a further decrease of – 30% in Wt mice after tMCAO (p = 0.02, F_(1,18) = 6.24). In FXII^?/? mice in which patency of the MCA was not restored (pMCAO) a further decrease of ? 75% was observed. Cortical reperfusion in tMCAO FXII^?/? mice was related to a lower risk of infarction of 59% vs. 93% in Wt mice (p = 0.04). Subcortical CBF was similarly decreased in both tMCAO groups (Wt and FXII^?/?) relating to a similar risk of infarction of 89% (Wt) vs. 99% (FXII^?/?, p = 0.17).Deficiency of FXII allows neocortical reperfusion after tMCAO and rescues brain tissue by this mechanism. This study supports the concept of FXII as a promising new target for stroke prevention and therapy.     

Clinical significance of Stratifin,ERα and PR promoter methylation in tumor and serum DNA in Indian breast cancer patients

Objectives:The objective of this study was to determine the concordance of promoter methylation of stratifin, ERα and PR in tumor and circulating DNA in breast cancer patients and their association with clinicopathological parameters and disease prognosis.Design and methods:Methylation specific PCR were carried out to investigate the promoter methylation status of stratifin, ERα and PR in tumor and circulating DNA in 100 breast cancer patients in a prospective study. The effect of promoter methylation on protein expression was evaluated by immunohistochemistry.Results:Significant association was observed between promoter methylation of stratifin in tumors (61%) and paired sera (56%) (r = 0.78; p ≤ 0.001). Loss of stratifin expression was observed in 47% tumors and was associated with poor overall survival (p = 0.05). Significant correlation was observed between methylation status of ERα with PRB (p < 0.0001, OR = 20.8, 95% CI = 7.4–58.0) and stratifin (p = 0.003, OR = 2.0, 95% CI = 0.8–4.4).Conclusion:This study underscores the potential utility of serum DNA methylation of these genes as surrogate for tumor DNA methylation as a promising tool for cancer diagnosis.     

Circulating osteoprotegerin is increased in the metabolic syndrome and associates with subclinical atherosclerosis and coronary arterial calcification

ContextThe relationship between osteoprotegerin (OPG) a glycoprotein related to bone metabolism and the metabolic syndrome (MS) has not been established.ObjectiveThe aim of this study is to evaluate OPG concentration in patients with MS and its association with subclinical atherosclerosis and coronary arterial calcification (CAC).Materials/methodsThe study included 238 asymptomatic patients. MS was diagnosed according to the NCEP/ATPIII guidelines. OPG was measured by ELISA. All subjects underwent ultrasonography of the common carotid arteries to measure intima-media thickness (IMT) and evaluate the presence of atheroma plaques. In a subgroup (n = 39) CAC was quantified by ECG-triggered cardiac computed tomography. Adipose tissue was excised from 25 patients and OPG expression by RT-PCR and immunohistochemistry was studied.ResultsPatients with the MS (n = 60) had higher OPG than patients without (n = 178) (p < 0.05). OPG correlated with IMT (r = 0.2, p = 0.005) and patients with atheroma plaques had higher OPG (p = 0.008) and also those with coronary artery calcification (p < 0.05).OPG expression was confirmed in adipose tissue (n = 12) and the expression was significantly higher in patients with MS than in those without (p = 0.003).ConclusionsThis study shows that OPG may potentially be a biomarker for cardiovascular risk/damage in the MS and identifies adipose tissue as a potential source of OPG.     

Novel role of Sarco/endoplasmic reticulum calcium ATPase 2 in development of colorectal cancer and its regulation by F36, a curcumin analog

Sarco/endoplasmic reticulum calcium ATPase (SERCA) enzymes play important roles in several signal transduction pathways that control proliferation, differentiation and apoptosis. Here, we reported that SERCA2 expression was positively correlated with tumor node metastasis (TNM) stages (n = 75, P = 0.0251) and grades (n = 63, P = 0.0146) of patients with colorectal cancer. The animal experiments demonstrated that SERCA2 expression was consistent with PCNA staining of intestinal tissues of male C57BL/6J-Apc^Min/JNju mice. Besides, SERCA2 expression was also increased in undifferentiated HT-29 cells as compared with that in differentiated HT-29 gal cells. Moreover, SERCA2 overexpression promoted proliferation and migration of SW480 cells via activating MAPK and AKT signaling pathways, while silence of SERCA2 inhibited the proliferation and migration of SW480 cells. In addition, we identified that a curcumin analog, F36, exhibited more potent inhibitory effect in colorectal cancer cells than curcumin through inhibiting SERCA2 expression. Taken together, our findings indicate that SERCA2 is involved in the malignant progress of colorectal cancer and maybe a therapeutic target for colorectal cancer treatment. Curcumin analog F36 shows enhanced anti-cancer activity in colorectal cancer cells by targeting SERCA2.     

Multi-center evaluation of a commercial Kit for tacrolimus determination by LC/MS/MS

ObjectivesA multi-center evaluation (3 sites) of the LC/MS/MS MassTrak? tacrolimus Immunosuppressants Kit (Kit) was undertaken.Design and methodsTen aspects of the analytical performance of the Kit were investigated based on FDA and CLSI guidelines.ResultsThe linear analytical range of the procedure was between 0.68 and 31.7 ng/mL. Within-run and total imprecision were < 6% and < 8% (n = 240), respectively. Recoveries of tacrolimus added to clinical samples that contained between 5 and 10 ng/mL of tacrolimus before addition were 99, 102 and 105% at 5.0, 10 and 20 ng/mL, respectively. Comparison of in-house and Kit procedures in samples from liver (n = 50–58) or kidney (n = 50 or 51) transplant recipients yielded method mean biases between ? 2.0 and + 10.7% at 5 and 15 ng/mL.ConclusionsThis evaluation indicates that the Kit is suitable for the monitoring of tacrolimus in kidney and liver transplant recipients.     

Predictive value of osteocalcin in bone metastatic differentiated thyroid carcinoma

ObjectivesTo evaluate the diagnostic value of serum osteocalcin in the detection of bone metastases from differentiated thyroid carcinoma (DTC).Design and methodsSerum samples from DTC patients with (DTC BM+, n = 19) or without bone metastases (DTC BM?, n = 19), and matched healthy volunteers (n = 30) were tested for serum osteocalcin with electrochemiluminescent immunoassay.ResultsOsteocalcin was higher in DTC BM+ than in DTC BM? patients (+ 35.8%, p = 0.002), acting as an independent risk factor for bone metastases (R² = 0.142, p = 0.039). The sensitivity was 78.9% and the specificity was 63.2% at a cut-off value of 11.2 μg/L.ConclusionsSerial measurements of osteocalcin could be useful in the detection of bone metastases from DTC.     

BSA-binding properties and anti-proliferative effects of amino acids functionalized polyoxomolybdates

Glycine decorated heteropolymolybdates, K₂Na[AsMo₆O₂₁(O₂CCH₂NH₃)₃]·6H₂O 1 and K₂Na₂[γ-Mo₈O₂₆(O₂CCH₂NH₃)₂]·6H₂O 2, have been synthesized and evaluated for in vitro anti-proliferative effects. The identity and high purity of compounds 1 and 2 were confirmed by elemental analysis, FT-IR spectrum, UV–vis spectrum, and X-ray diffraction. Crystal data for 2: triclinic, P-1, a = 9.792(2) Å, b = 10.077(2) Å, c = 10.351(2) Å, α = 83.865(4)°, β = 71.110(4)°, γ = 62.284(3)°, V = 854.3(3) Å³, Z = 1, R(final) = 0.0486. The inhibitory effects of 1 and 2 on human non-small cell lung cancer cell line A549 were investigated by MTT assay, nuclear staining, and the flow cytometry. It indicated that compound 1 inhibited the proliferation of A549 cells in a dose-dependent and time-dependent manner, which is more effective than the positive control, 5-fluorouracil (5-FU) (P < 0.05). The staining and flow cytometry results showed that compound 1 induced the apoptosis and necrosis of A549 cells and inhibit cell proliferation, which is associated with S-phase arrest. Compound 2 showed a modest activity in a dose-dependent manner. In addition, the interaction between compound 1 and bovine serum albumin (BSA) was evaluated by spectroscopic methods. The results showed that the compound 1 effectively quenched the intrinsic fluorescence of BSA via static quenching and changed the conformation of BSA.     

Kallikrein-related peptidase 13 (KLK13) gene expressional status contributes significantly in the prognosis of primary gastric carcinomas

ObjectivesGastric cancer is a fatal human malignancy with poor prognosis. Modifications in gene expression, including those of the kallikrein-related peptidase family, have been portrayed in gastric carcinogenesis. Given KLK13 involvement in human malignancies, we aimed to uncover its prognostic strength in stomach cancer.Design and methodsQuantitative analysis of KLK13 profiles was accomplished in human gastric cancer cells and in a statistically significant sample size of stomach tissue specimens with the development of the highly sensitive real-time PCR methodology.ResultsDecreased KLK13 expression was demonstrated in cancerous compared with their matching non-malignant pairs (p = 0.002) and in poorly differentiated gastric tumors (p = 0.029). KLK13-positive patients were shown to live considerably longer (p = 0.014) and with low risk of disease recurrences (p = 0.043).ConclusionsThis is the first study disclosing the possible clinical utility of KLK13 as a new tumor biomarker capable of predicting a favorable outcome for gastric cancer patients.     

Soluble urokinase-type plasminogen activator receptor forms in plasma as markers of atherosclerotic plaque vulnerability

Objectives:To test if circulating forms of the soluble urokinase-type plasminogen activator receptor (suPAR) are potential biomarkers of plaque vulnerability.Design and methods:Plasma concentrations of suPAR(I–III), suPAR(II–III) and uPAR(I) were measured by time-resolved fluorescence immunoassays in Caucasian patients operated for symptomatic carotid atherosclerosis (n = 255). Local suPAR release from plaques into the circulation was assessed in plasma passing retrogradely over the plaque in the carotid artery, collected during surgery (n = 7).Results:The suPAR(I–III) (P = 0.03) and suPAR(II–III) (P = 0.006) concentrations were higher after ischemic strokes and transient ischemic attacks, i.e., clinical subgroups associated with poorer prognosis and a less stable plaque phenotype, than after amaurosis fugax. Slightly elevated suPAR(I–III) levels were found in plasma from the carotid lesion. However, refuting the hypothesis, the concentrations of the suPAR forms were not higher in patients with short intervals between clinical event and blood sampling compared with those with long intervals. Age, inflammatory markers and diabetes were confounding factors independently associated with suPAR forms.Conclusion:Circulating suPAR forms are probably not useful biomarkers of atherosclerotic plaque vulnerability.     

Video laryngoscopy versus direct laryngoscopy for tracheal intubation during in-hospital cardiopulmonary resuscitation

AimTracheal intubation during cardiopulmonary resuscitation (CPR) is a high-risk procedure. Here, we investigated the efficacy of video laryngoscopy for tracheal intubation during CPR.MethodsData regarding tracheal intubation during CPR from in-hospital cardiac arrests occurring between January 2011 and December 2013 (n = 229) were prospectively collected and retrospectively analyzed.ResultsThe initial laryngoscopy method was video laryngoscopy in 121 patients (52.8%) and direct laryngoscopy in 108 patients (47.2%). The rate of successful intubation at the first attempt was higher with video laryngoscopy (71.9%; 87/121) than with direct laryngoscopy (52.8%; 57/108; p = 0.003). The rate of success at the first attempt was higher for experienced (73.0%; 84/115) than inexperienced operators, including residents (52.6%; 60/114; p = 0.001). Mortality at day 28 after CPR was not significantly different between patients with successful tracheal intubation at the first attempt and without (68.1% [98/144] vs. 67.1% [57/85]; p = 0.876). In multivariate logistic regression analysis, a predicted difficult airway (odds ratio [95% confidence interval] = 0.22 [0.10–0.49]; p < 0.001), intubation by an experienced operator (2.63 [1.42–4.87]; p = 0.002), and use of video laryngoscopy rather than direct laryngoscopy (2.42 [1.30–4.45]; p = 0.005) were independently associated with a successful tracheal intubation at the first attempt.ConclusionUse of video laryngoscopy during CPR from in-hospital cardiac arrest is independently associated with successful tracheal intubation at the first attempt.     

Management of Moderate-to-Severe Dyspnea in Hospitalized Patients Receiving Palliative Care

ContextBenzodiazepines (BZDs) are commonly prescribed for relief of dyspnea in palliative care, yet few data describe their efficacy.ObjectivesTo describe the management of moderate-to-severe dyspnea in palliative care patients.MethodsChart review of inpatients with moderate or severe dyspnea on initial evaluation by a palliative care service. We recorded dyspnea scores at follow-up (24 hours later) and use of BZDs and opioids.ResultsThe records of 115 patients were reviewed. The mean age of patients was 64 years and primary diagnoses included cancer (64%, n = 73), heart failure (8%, n = 9), and chronic obstructive pulmonary disease (5%, n = 6). At initial assessment, 73% (n = 84) of the patients had moderate and 27% (n = 31) had severe dyspnea. At follow-up, 74% (n = 85) of patients reported an improvement in their dyspnea, of which 42% (n = 36) had received opioids alone, 37% (n = 31) had BZDs concurrent with opioids, 2% (n = 2) had BZDs alone, and 19% (n = 16) had received neither opioids nor BZDs. Logistic regression analysis identified that patients who received BZDs and opioids had increased odds of improved dyspnea (odds ratio 5.5, 95% CI 1.4, 21.3) compared with those receiving no medications.ConclusionMost patients reported improvement in dyspnea at 24 hours after palliative care service consultation. Consistent with existing evidence, most patients with dyspnea received opioids but only the combination of opioids and BZDs was independently associated with improvement in dyspnea. Further research on the role of BZDs alone and in combination with opioids may lead to better treatments for this distressing symptom.     

Urinary matrix metalloproteinase-8 and -9 activities in type 2 diabetic subjects: A marker of incipient diabetic nephropathy?

Matrix metalloproteinases (MMPs) may play a pathophysiological role in the development of diabetic nephropathy (DN). We hypothesized that urinary MMP activity in patients with type 2 diabetes mellitus (T2DM) is related to a decline in renal function. We determined MMP-2, -8 and -9 activity in 24-h urine collections in relation to risk factors for DN in T2DM patients with (UA, n = 27) and without albuminuria (NA, n = 48) and controls (CO, n = 28). MMP-8 and -9 levels were highest in UA patients (P < 0.01). Of UA patients, 93% had at least one MMP increased, compared to 78% of NA patients and 46% of CO (P = 0.001). Age, diabetes duration, BMI, systolic blood pressure, fasting plasma glucose, HbA1c and renal function were determinants of MMP-8 and -9 (P < 0.05). In summary, MMP-8 and -9 are highest in T2DM UA patients. MMP-9, showed the strongest associations with clinical parameters related to DN.