Endostatin gene therapy inhibits intratumoral macrophage M2 polarization

BackgroundRenal cell carcinoma (RCC) is a highly vascularized cancer resistant to chemotherapy and radiotherapy. RCC is frequently infiltrated with immune cells, with macrophages being the most abundant cell type. Alternatively activated M2 macrophages are known to contribute to tumor progression. Endostatin (ES) is a fragment of collagen XVIII that possesses antiangiogenic activity. In this study, we investigated the impact of ES gene therapy on the polarization of tumor-associated macrophages (TAMs) in lung metastases from tumor-bearing mice.MethodsBALB/c mice divided into three groups: Normal, Control and ES-treated. Tumor-bearing mice were treated with ES-transduced cells or control cells over ten days. At the end of the study, plasma was collected, and pulmonary macrophages were isolated and used for FACS or RT-PCR. ELISA tests were used to analyze plasma and cell culture supernatant cytokines.ResultsES treatment significantly reduced the levels of anti-inflammatory and pro-angiogenic cytokines, including IL4, IL-10, IL-13 and VEGF. Gene expression of M2 markers, such as IL-10, Arg-1, VEGF and YM-1, declined significantly. Flow cytometry showed a reduction in the number of M2 F4/80 + CD36 + CD206 + CD209+ macrophages and in IL-10 secretion by these cells. Reduced levels of IL-10 were also found in the culture supernatants of the ES-treated group.ConclusionsOur research corroborates previous observations that ES has an important anti-tumoral role. However, aside from promoting interferon-ɤ secretion and an effective T cell response, we show here that this switch is extended to TAMs, complicating the maintenance of pro-tumorigenic M2 macrophages and thus favoring tumor elimination.     

Increased uptake of oxidized LDL by macrophages from type 2 diabetics is inhibited by polyamines

The aim of this study was to evaluate the effect of polyamines putrescine, spermidine and spermine on human LDL oxidation and to assess the ability of macrophages derived from type 2 diabetic patients to uptake oxLDL. Polyamine effect was compared with α-tocopherol. Four healthy subjects and eight type 2 diabetic patients were included in this study. To characterize type 2 diabetic patients and non-diabetic subjects, laboratory test were carried out. Glucose, glycated haemoglobin (HbA_1C), triglycerides, low (LDL) and high density lipoproteins (HDL) and serum lipid peroxidation were measured in blood. The study was performed in three stages. For each stage, ten experimental conditions comparing the effect of polyamines with α-tocopherol (10 μM solutions) on LDL oxidation and the uptake of oxLDL by macrophages were analyzed. MDA concentration was found to be significantly higher in type 2 diabetic patients compared to healthy subjects (5.6 ± 0.58 vs. 2.66 ± 0.31 μM MDA, respectively, (P < 0.05)). Percent of macrophages containing oxLDL was determined by means of red oil staining. The uptake of oxLDL by macrophages derived from diabetic patients was clear. The uptake of oxLDL was inhibited when the oxidation was prevented by polyamines or α-tocopherol. Spermine showed high antioxidant capacity (96.67 ± 1.53% vs. 25.67 ± 2.30%) compared to α-tocopherol (96.67 ± 1.53% vs. 47.00 ± 7.20%) at the concentration tested.In conclusion, polyamines especially spermine, has a potent antioxidant effect compared to α-tocopherol on human LDL oxidation, followed by spermidine and putrescine. The results have clinical relevance in the diabetic complications and add knowledge on the role of polyamines as natural antioxidants. This research is not a clinical evaluation rather a functional analysis utilizing clinical samples.     

Alteration of HDL functionality and PON1 activities in acute coronary syndrome patients

ObjectiveThe functionality of HDL has been suggested as an important factor in the prevention of cardiovascular and coronary artery diseases. The objective of the present study was to investigate the functionality of HDL and the factors that may affect the anti-atherogenic properties of HDL in ACS patients.Methods and resultsOne hundred healthy subjects and 205 ACS patients were recruited. HDL functionality was evaluated by measuring their capacity to mediate cholesterol efflux from J774 macrophages. Oxidative stress status was determined by measuring plasma malondialdehyde (MDA), protein carbonyl, and vitamin E levels by HPLC. The PON1 Q192R polymorphism status and PON1 paraoxonase and arylesterase activities of the healthy subjects and ACS patients were also determined. The HDL of ACS patients displayed a limited capacity to mediate cholesterol efflux, especially via the ABCA1-pathway. MDA (7.06 ± 0.29 μM) and protein carbonyl (9.29 ± 0.26 μM) levels were significantly higher in ACS patients than in healthy subjects (2.29 ± 0.21 μM and 3.07 ± 0.17 μM, respectively, p < 0.0001), while α- and γ-tocopherol (vitamin E) levels in ACS patients were 8-fold (p < 0.001) and 2-fold (p < 0.05) lower than in healthy subjects. Paraoxonase, arylesterase and HDL-corrected PON1 activities (PON1 activity/HDL ratio) were significantly lower in ACS patients. Logistic regression analyses showed that high PON1 paraoxonase and arylesterase activities had a significant protective effect (OR = 0.413, CI 0.289–0.590, p < 0.001; OR = 0.232 CI 0.107–0.499, p < 0.001, respectively) even when adjusted for HDL level, age, BMI, and PON1 polymorphism.ConclusionThe results of the present study showed that the functionality of HDL is impaired in ACS patients and that the impairment may be due to oxidative stress and an alteration of PON1 activities.     

Mild mental retardation and low levels of urinary heparan sulfate in a patient with the attenuated phenotype of mucopolysaccharidosis type IIIA

ObjectivesWe report the case of a 28-year-old female subject affected by the attenuated phenotype of mucopolysaccharidosis type IIIA characterized by moderate slowly evolving mental retardation in which the urinary content of heparan sulfate was demonstrated as being substantially low compared to that found in patients with the severe phenotype.Design and methodsThe specific evaluation of macromolecular heparan sulfate by electrophoresis and the determination of related glucosamine in the urine were performed.ResultsIn our patient, the urinary macromolecular heparan sulfate content (4.2 μg/mg creatinine) was ~ 7.5-times higher than in healthy subjects (0.56 μg/mg creatinine ± 0.9 SD) while it was ~ 28-times lower compared to the severe mucopolysaccharidosis IIIA group (117 μg/mg creatinine ± 44.8 SD). Furthermore, the urinary glucosamine (86.4 μg/mg creatinine) was ~ 2.4-times greater than in healthy subjects (36.0 μg/mg creatinine ± 18.2 SD) but ~ 2.4-times lower than in severe subjects (208.1 μg/mg creatinine ± 55.0 SD).ConclusionsThe above data could reflect the reduced heparan sulfate storage in her tissues and organs, and in particular in the brain, consequently explaining her moderate mental retardation. Furthermore, the clinical presentation of patients with an attenuated form of MPS III confirms the need for a specific evaluation of urinary GAGs in all young and adult subjects showing a not well-defined or not particularly severe mental retardation, along with an early MPS diagnosis. Such investigation should also be associated with a more specific characterization of heparan sulfate.     

Glutamate dehydrogenase activity in lymphocytes of B-cell chronic lymphocytic leukaemia patients

Objectives: To investigate the pattern of glutamate dehydrogenase (GLDH) activity, GLUD1 and GLUD2 expressions in peripheral blood mononuclear cells (PBMC) of untreated B-chronic lymphocytic leukemia (B-CLL) in healthy individuals (HI) and patients with infectious mononucleosis (IM).Design and methods: GLDH activity was determined in a supernatant obtained from pelleted PBMC. GLUD1 and GLUD2 mRNA expression was determined using a quantitative real-time polymerase chain reaction. CD19⁺ B cells from PBMC were purified by using positive selection.Results: The highest GLDH activity was found in PBMC of the B-CLL group followed by the HI group and IM group. The PBMC GLDH activity was higher in 60% of the B-CLL patients according to the established reference interval for our HI (2.17–5.70 μkat/g protein). The greater GLDH activity was also found in the CD19⁺ cell preparation of the B-CLL patients (two of the three) but not in HI (n = 3). The median value of GLUD1 expression was highest in the IM group (n = 11), followed by the HI (n = 14) and B-CLL groups (n = 59) (median 4.69/3.78, P < 0.005 and 4.69/2.91, P < 0.0005, respectively). GLUD2 expression was not significantly different between groups.Conclusions: The increased GLDH activity is specific for the PBMC of B-CLL patients. The GLUD1 but not the GLUD2 gene expression pattern is different between the PBMC of IM and B-CLL patients.     

Effects of V279F in the Lp-PLA2 gene on markers of oxidative stress and inflammation in Koreans

BackgroundA single nucleotide polymorphism (SNP), V279F, in the lipoprotein-associated phospholipase A₂ (Lp-PLA₂) gene is known to influence enzyme activity. It is unclear whether Lp-PLA₂ exerts pro- or antiatherogenic effects in humans. We investigated the interplay between V279F, Lp-PLA₂ activity, oxidative stress and inflammation.MethodsWe genotyped 2914 healthy Koreans (43–79 years) for the Lp-PLA₂ V279F and measured anthropometric parameters, lipid profile, fatty acid composition, lipid peroxides, inflammatory markers and Lp-PLA₂ levels.ResultsLp-PLA₂ activity was 24% lower in V/F subjects (n = 641) than in those with the V/V genotype (n = 2227). Enzyme activity was undetectable in F/F subjects. Lp-PLA₂ activity was positively correlated with LDL-cholesterol (r = 0.134, P < 0.001), ox-LDL (r = 0.064, P < 0.01), 8-epi-PGF_2α (r = 0.198, P < 0.001), free fatty acid (r = 0.082, P < 0.001), and fibrinogen (r = 0.112, P < 0.01) levels. Additionally, ox-LDL, 8-epi-PGF_2α, free fatty acid, and fibrinogen levels were positively correlated with hs-CRP. V279F was associated with LDL-cholesterol and arachidonic acid (AA) in serum phospholipid. F/F subjects had lower LDL-cholesterol than V/V subjects (V/V: 120.9 ± 0.69, V/F: 119.4 ± 1.26, F/F: 109.2 ± 4.84 mg/dl, P = 0.025). A significant association between the F/F genotype and increasing AA in serum phospholipids was found in subjects with high LDL-cholesterol (≥ 130 mg/dl) (P = 0.003) but not in those with low LDL-cholesterol (< 130 mg/dl). F/F subjects in the high LDL-cholesterol group had CRP concentrations about three times higher than those with V/V or V/F genotypes (V/V: 1.25 ± 0.09, V/F: 0.97 ± 0.12, F/F: 3.20 ± 0.88 mg/dl, P < 0.001).ConclusionsThe recessive effects of Lp-PLA₂ V279F on LDL-cholesterol and significant correlations between Lp-PLA₂ activity and LDL-cholesterol, 8-epi-PGF_2α and fibrinogen support a pro-oxidative or pro-atherogenic role for this enzyme. Paradoxically, the combination of the complete deficiency of Lp-PLA₂ activity and high LDL-cholesterol enhanced lipid peroxidation and inflammation.     

Improved cisplatin delivery in cervical cancer cells by utilizing folate-grafted non-aggregated gelatin nanoparticles

PurposeCisplatin is highly effective in the treatment of cervical cancer. However, in therapeutic doses, cisplatin induces several adverse effects due to undesirable tissue distribution. Therefore, it is worth targeting cisplatin in cervical cancer cells by implicating non-aggregated ligand-modified nanotherapeutics.Methods and resultsHere, we report the preparation of non-aggregated folic acid-conjugated gelatin nanoparticles of cisplatin (Cis-GNs-FA) by two-step desolvation method with mean particle size of 210.6 ± 9.6 nm and 140.5 ± 10.9 nm for Cis-GNs to improve the drug delivery in cervical cancer, HeLa cells. FTIR and DSC spectra confirmed the presence and stability of cisplatin in gelatin matrix. Furthermore, amorphization of cisplatin in nanoparticles was ascertained by PXRD. Drug release followed a first-order release kinetic at both pH ∼ 5.6 (cervical cancer pH) and pH ∼ 7.4. In addition, a significant (P < 0.05) decrease in IC₅₀ value (8.3 μM) and enhanced apoptosis were observed in HeLa cells treated with Cis-GNs-FA as compared to Cis-GNs (15.1 μM) and cisplatin solution (40.2 μM). In contrast, A549 lung cancer cells did not discriminate between Cis-GNs-FA and Cis-GNs due to the absence of folate receptors-α (FR-α). Consistently, higher cellular uptake, 80.54 ± 7.60% was promoted by Cis-GNs-FA significantly (two-way ANOVA, P < 0.05) greater than 51.68 ± 9.78%, by Cis-GNs. This was also illustrated by CLSM images, which indicated that Cis-GNs-FA preferably accumulated in the cytoplasm of HeLa cells nearby nucleus by following receptor-mediated endocytosis pathway as compared to Cis-GNs.ConclusionTherefore, Cis-GNs-FA warrants further in-depth in vitro and in vivo investigations to scale up the technology for clinical translation.     

Synthesis and photodynamic activity of unsymmetrical A3B tetraarylporphyrins functionalized with l-glutamate and their Zn(II) and Cu(II) metal complex derivatives

Four novel unsymmetrical A₃B porphyrins 1, 2, 3 and 4 were synthesized following Lindsey procedure. Porphyrins 3 and 4 include one and three l-glutamate groups, respectively, and all porphyrins were metallated with Zn(II) (1a–4a) or Cu(II) (1b–4b). Porphyrins and metalloporphyrins presented values of singlet oxygen quantum yields (ΦD) ranging from 0.21 to 0.67. The tetraaryl derivatives in this study showed phototoxicity in SiHa cells with IC₅₀ values ranging from <0.01 to 6.56 ± 0.11 μM, the metalloporphyrin 4a showed the lowest IC₅₀ value. Comparing the phototoxic activity between all porphyrins, functionalization of porphyrins with glutamate increased 100 times phototoxic activity (1 (IC₅₀ 4.81 ± 0.34 μM) vs. 3 (IC₅₀ 0.04 ± 0.02 μM) and 2 (IC₅₀ 5.19 ± 0.42 μM) vs. 4 (IC₅₀ 0.05 ± 0.01 μM)). This increased activity could be attributed to reduced hydrophobicity and increased ΦΔ, given by functionalization with l-glutamate. Metalloporphyrins 3a (IC₅₀ 0.04 ± 0.01 μM) and 4a (IC₅₀ <0.01 μM) presented the best values ​​of phototoxic activity. Therefore, functionalization and zinc metalation increased the phototoxic activity. SiHa cells treated with porphyrins 3, 4, 3a and 4a at a final concentration of 10 μM, showed increased activity of caspase-3 enzyme compared to the negative control; indicating the induction of apoptosis. Differential gene expression pattern in SiHa cells was determined; treatments with metalloporphyrins 4a and 4b were performed, respectively, comparing the expression with untreated control. Treatments in both cases showed similar gene expression pattern in upregulated genes, since they share about 25 biological pathways and a large number of genes. According to the new photophysical properties related to the structural improvement and phototoxic activity, these molecules may have the potential application as photosensitizers in the photodynamic therapy.     

Comparative evaluation of neonatal bilirubin using Radiometer whole blood co-oximetry and plasma bilirubin methods from Roche Diagnostics and Ortho Clinical Diagnostics

BackgroundClinically significant variation has been reported within and between plasma and whole blood total bilirubin methods used to identify neonates for whom clinical intervention for hyperbilirubinemia may be required.ObjectiveTo evaluate total bilirubin measurements between the Radiometer whole blood co-oximeter and plasma bilirubin methods from Roche Diagnostics and Ortho Clinical Diagnostics using neonatal specimens.MethodsTotal bilirubin levels were analyzed by whole blood co-oximetry (Radiometer® ABL90). Specimens were centrifuged and plasma analyzed for total bilirubin with a diazo method (Roche Cobas® C-601) and a reflectance spectrophotometric BuBc dry film method (Ortho Clinical Diagnostics VITROS® 350). Results were evaluated by regression, Bland-Altman comparisons and t-tests.ResultsThe patient correlation study yielded the following regression equations in μmol/L: a) Radiometer = 1.03 Roche – 3.5 μmol/L b) Radiometer = 0.98 Ortho – 5.7 μmol/L c) Roche = 0.97 Ortho – 2.4 μmol/L. The mean bias over the range of total bilirubin levels examined was − 1.0 μmol/L for the Radiometer versus the Roche (p ≤ 0.305); − 4.4 μmol/L for the Radiometer versus Ortho (p ≤ 0.005) and − 4.4 μmol/L for the Roche versus Ortho (p ≤ 0.002).ConclusionsWhole blood total bilirubin measurement using the Radiometer ABL90 blood gas analyzer provides accurate and precise results compared to the Roche plasma diazo method. Compared to the reflectance spectrophotometric method, results are precise and had a small but statistically significant bias of − 4.4 μmol/L.     

Atrophy of individual thigh muscles measured by MRI in older adults with knee osteoarthritis: A cross-sectional study

BackgroundThe characteristics of thigh-muscle cross-sectional area (CSA) in older adults with knee osteoarthritis (KOA) remain controversial.ObjectivesThis study aimed to evaluate atrophy of individual thigh muscles in older adults with KOA and to determine which muscle CSA should be measured to detect KOA-related muscle atrophy of the thigh.MethodsIn older adults, individual thigh-muscle CSA measured by 1.5 Tesla MRI was analyzed at 5% intervals of the femoral length (FL) around the mid-thigh between the proximal 25% of the FL and the distal 25%. Participants with KOA grade ≤ 1 and grade ≥ 2 were compared for ratios of quadriceps muscle (QM) CSA to total thigh, individual QM CSA to QM, and individual hamstring (HAM) CSA to HAM at 5% intervals.ResultsWe included 40 older adults [20 males; mean (SD) age 73.3 (4.7) years; 20 with KOA grade ≤ 1 and 20 with KOA grade ≥ 2]. The ratio of vastus medialis (VM) CSA to QM from the proximal 25% to distal 15% and the ratio of semi-membranosus (SM) CSA to HAM at the distal 10% to 25% were significantly lower with KOA grade ≥ 2 than grade ≤ 1; the effect sizes were 0.34 to 0.67 for VM and 0.40 to 0.60 for SM. The effect sizes were greatest for the ratios of VM CSA to QM at the mid-thigh with 5% intervals and the ratio of SM CSA to HAM at the distal 25%.ConclusionsThe ratio of VM CSA to QM and/or that of SM CSA to HAM were low and were the best indicators to detect KOA-related muscle atrophy of the thigh. However, to detect KOA-related muscle atrophy, the VM CSA ratio should be analyzed in the thigh region around the mid-thigh, whereas the SM CSA ratio should be analyzed in the thigh region at the muscle belly.     

Role of NF-E2-related factor 2 in neuroprotective effect of l-carnitine against high glucose-induced oxidative stress in the retinal ganglion cells

l-Carnitine (LC) has protective effects on high glucose-induced oxidative stress in the retinal ganglion cells (RGCs). The aim of this study was to investigate the role of NF-E2-related factor 2 (Nrf2), Kelch like-ECH-associated protein 1 (Keap1), haemoxygenase-1 (HO-1) and γ-glutamyl cysteine synthetase (γ-GCS) in the protective effect of LC on RGCs. RGCs were first processed with high concentrations of glucose. LC treatment at three concentrations (50 μM, 100 μM and 200 μM) was applied to high glucose stimulated RGCs. The expression of Nrf2, Keap1, haemoxygenase-1 (HO-1) and γ-glutamyl cysteine synthetase (γ-GCS) was quantified by Western blot in the treatment and control (high glucose stimulation) groups. In the three LC groups (50 μM, 100 μM and 200 μM), Nrf-2 (0.71 ± 0.04, 0.89 ± 0.05, 1.24 ± 0.05 vs 0.56 ± 0.03, p < 0.05), HO-1 (0.58 ± 0.04, 0.76 ± 0.06, 0.89 ± 0.07 vs 0.25 ± 0.03, p < 0.01), and γ-GCS protein expression (0.66 ± 0.03, 0.79 ± 0.05, 0.84 ± 0.08 vs 0.84 ± 0.08, p < 0.01) was higher than in the control group. The levels of Keap1 protein were in the LC groups were lower than in the control group (0.50 ± 0.03, 0.45 ± 0.02, 0.53 ± 0.03 vs 0.86 ± 0.05, p < 0.01). In conclusion, in high glucose stimulated RGCs, LC treatment was associated with an increased level of Nrf2, HO-1and γ-GCS. LC treatment was also associated with a reduced expression of Keap1 protein. These results suggest that the protective effect of LC treatment on RGCs may be related to Nrf2-Keap1 pathway.     

Verification of an immunoturbidimetric assay for heart-type fatty acid-binding protein (H-FABP) on a clinical chemistry platform and establishment of the upper reference limit

BackgroundHeart-type fatty acid-binding protein (H-FABP) is an early biomarker of cardiac injury. Randox Laboratories developed an immunoturbidimetric H-FABP assay for non-proprietary automated clinical chemistry analysers that could be useful in the emergency department. We verified the analytical performances claimed by Randox Laboratories on Roche Cobas 6000 clinical chemistry platform in use in our laboratory, and we defined our own 99th percentile upper reference limit for H-FABP.MethodsFor the verification of method performances, we used pools of spared patient samples from routine and two levels of quality control material, while samples for the reference value study were collected from 545 blood donors. Following CLSI guidelines we verified limit of blank (LOB), limit of detection (LOD), limit of quantitation (LOQ), repeatability and within-laboratory precision, trueness, linearity, and the stability of H-FABP in EDTA over 24 h.Results and discussionThe LOQ (3.19 μg/L) was verified with a CV% of 10.4. The precision was verified for the low (mean 5.88 μg/L, CV = 6.7%), the medium (mean 45.28 μg/L, CV = 3.0%), and the high concentration (mean 88.81 μg/L, CV = 4.0%). The trueness was verified as well as the linearity over the indicated measurement interval of 0.747–120 μg/L. The H-FABP in EDTA samples is stable throughout 24 h both at room temperature and at 4 °C. The H-FABP 99th percentile upper reference limit for all subjects (3.60 μg/L, 95% CI 3.51–3.77) is more appropriate than gender-specific ones that are not statistically different.     

Scavenging properties of neutrophil 4-hydroxyphenylpyruvate dioxygenase are based on a hypothesis that does not stand up to scrutiny

It was previously reported by D’Eufemia et al. [9] that neutrophil preparations from a patient with tyrosinemia type III, i.e. with inherited deficiency of 4-hydroxyphenylpyruvate dioxygenase (HPPD), exhibited a far higher NO release than controls, when NO was estimated in terms of nitrite content in the suspending media. It was hypothesized that HPPD might participate to NO sequestration in neutrophils and that excessive NO release might reflect the lack of the scavenging action in defective cells. In recent control experiments, we found that HPPD activity in neutrophils preparations from healthy subjects is below the detection limit of the enzymatic assay (less than 3 nmol product/h per mg protein). This indicates that HPPD concentration in neutrophils is very low, if any, confirming what was already suggested in literature, and rules out the possibility of a prominent role of HPPD as NO scavenger in these cells. Moreover, we found that 500 μM l-tyrosine increases nitrite release and accumulation in suspending media of U-937 cells, a human monoblast-like lymphoma cell line which displays many characteristics of macrophages, including the expression of inducible and endothelial nitric oxide synthases. We hypothesize that the increase of nitrite release by patient's neutrophils might be related to the presence of high l-tyrosine concentrations in the blood samples (426 μmol/L instead of 52.1 ± 10.9 μmol/L as healthy subjects), rather than to HPPD deficiency of in these cells.     

Hypervariability in a minisatellite 3' of the apolipoprotein B gene: Allelic distribution and influence on lipid profiles in Han Children from central China

BackgroundApolipoprotein B (apoB) gene 3' variable number tandem repeat (VNTR) is highly variable, and thereby be considered as an informative marker for associative analysis of lipid metabolism.MethodsWe conducted this study to probe the effect of apoB 3' VNTR alleles on lipid profiles in 500 Han children from central China, and to compare the allelic distribution of our subjects with multiple Chinese populations. 14 different alleles of the apoB gene 3' VNTR comprising from HVE22 to HVE44 were identified in our subjects.ResultsAllele size distribution followed unimodal curve with the main peak at HVE35 (58.0%). We detected 37 genotypes in this sampling, the most frequently seen was HVE35/35 with a frequency of 36.4%. M/L carriers had significantly higher total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C) and apoB concentrations than did S/S, M/M or S/M carriers (p < 0.05). Individuals with L allele exhibited significantly higher TC, LDL-C, and apoB levels than those with M or S allele (p < 0.05). The allelic distribution in Central Han Chinese differed from Southern Han Chinese (X² = 41.2, p = 0.00), Zhuang Chinese (X² = 65.4, p = 0.00), and Uighur Chinese (X² = 45.6, p = 0.00). No significant differences in allelic frequencies were observed for apoB 3' VNTR in Central Han Chinese as compared to Northern Han Chinese (X² = 2.5, p = 0.29).ConclusionThis study identified the higher repeat alleles as potential risk factor for dyslipidemia in Han children from Central China. Although five Chinese populations demonstrated uniformly unimodal distributions of allelic frequencies with the main peaks at HVE32–HVE37, there was obvious heterogeneity among these populations.     

Vitamin E (α-tocopherol) enhances the PDT action of hematoporphyrin derivatives on cervical cancer cells

ObjectiveThe effect of antioxidant vitamin E (VE) on PDT (a mainly reactive oxygen species-driven process) has in the past shown contradicting results. Hence, we studied the effect of different concentrations and different incubation periods of VE on the PDT cytotoxicity of the cervical adenocarcinoma HeLa cell line.Materials and methodsHeLa cells were incubated with 25 μg/ml of hematoporphyrin derivatives (HpD) for 25 min (1st PDT regimen) or 24 h (2nd PDT regimen), then irradiated with visible light (total light dose of 10 J/cm²) either with or without different concentrations of VE (1–1000 μM) which had been incubated with the cells for 1 h or 24 h prior to PDT. After irradiation, viability was measured using MTT assay.ResultsThe results obtained showed that PDT is effective against cervical cancer cells. Incubation of HpD for 24 h leads to improved PDT action. Higher concentrations of VE incubated in HeLa cells for 1 h before the 2nd PDT regimen significantly enhanced cytotoxicity of PDT and the maximal enhancement was at 1000 μM of VE. The cytotoxic effect of VE on HeLa cells after incubation for 24 h before PDT is enhanced by the PDT action.ConclusionIn conclusion 1000 μM of VE can be used 1 h before PDT to enhance its effect on cervical adenocarcinoma, a disease which is steadily increasing in young women. It is well-known that the cervical adenocarcinoma is resistant to anticancer agents and radiotherapy and it was previously considered to be HpD-PDT resistant.     

Impact of long distance rowing on biological health: A pilot study

ObjectivesTo determine the impact of long distance rowing (160 km, nonstop) on standard biological parameters and to study the relation between inflammation, myocardial necrosis, lipid profile, heart rate and energy expenditure.MethodsElectrolytes, lipid profile, C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), procalcitonin (PCT), high-sensitive troponin T (hs-cTnT), and N-terminal pro-brain natriuretic peptide (NT-proBNP), were measured on non-fasting venous blood samples collected 8 h before and after the rowing race on five healthy competitors. Heart rate and energy expenditure were measured using sporting self-measurement devices.ResultsAfter 16.5 h of race, significant increases in median CRP (+ 25.2 mg/l; p = 0.04), IL-6 (+ 1.85 pg/ml; p = 0.04), TNF-α (+ 1.2 pg/ml; p = 0.04) and NT-proBNP levels (+ 88.8 pg/ml; p = 0.04) were observed, and a close to significant elevation for hs-cTnT(+ 6 ng/l; p = 0.06) and PCT (+ 0.14 μg/l; p = 0.07). On the other hand, significant decrease in median total cholesterol (− 0.5 mmol/l; p = 0.04), triglycerides (− 0.7 mmol/l; p = 0.04) were observed. Furthermore, significant correlations between the maximal heart rate reached during the race and CRP (r = 0.90; p = 0.03), IL-6 (r = 0.90; p = 0.03), and NT-proBNP (r = 0.90; p = 0.03) were observed, whereas no such associations were retrieved with median heart rate, the percentage of time passed over 70% of maximal heart rate or energy expenditure during the race. There was no association between PCT, NT-proBNP, hs-cTnT, inflammatory biomarkers, lipid profile or heart rate parameters.ConclusionsLong distance rowing induces inflammation and myocardial strain related to the maximal effort generated during the race, but has a favourable effect on lipid profile.     

The clearance time of infused hematopoietic stem cell from the blood circulation

There is no detailed information about the clearance time of infused hematopoietic stem cell (HSC) from the blood circulation in humans. In this prospective study, peripheral blood CD34+ cell counts were detected during the 4 days period following autologous HSC transplantation in 20 patients by means of flow cytometry. The median CD34+ cells were at the highest level in the first hour and decreased below pre-infusion values on the first day after HSC infusion. By nonparametric analysis, positive correlation was found between CD34+ cell levels at the first hour and the post-thaw CD34+ cell dose (r = 0.57, p = 0.01). An inverse correlation was determined between CD34+ cell levels at the first hour and neutrophil engraftment (r = ?0.54, p = 0.01). Compared with the patients having CD34+ cell count of ?2 μL^?1 in the first hour following HSC infusion, the patients having CD34+ cell count of <2 μL^?1 had delayed both neutrophil (20 vs. 12, p = 0.008) and platelet (47 vs. 11, p = 0.01) engraftments. Our results indicated that infused HSCs were removed from the blood circulation within 1 day. In addition, CD34+ cell levels at the first hour may be used as an important indicator to predict the delay of neutrophil and platelet engraftments.     

Effects of curcumin on serum cytokine concentrations in subjects with metabolic syndrome: A post-hoc analysis of a randomized controlled trial

BackgroundCytokines are involved in the development of metabolic abnormalities that may result in metabolic syndrome (MetS). Since curcumin has shown anti-inflammatory properties, the aim of this study was to evaluate the effect of curcumin supplementation on serum cytokines concentrations in subjects with MetS.MethodsThis study was a post-hoc analysis of a randomized controlled trial in which males and females with diagnosis of MetS, according to the criteria defined by the National Cholesterol Education Program Adult Treatment Panel III guidelines, were studied. Subjects who met the inclusion criteria were randomly assigned to either curcumin (daily dose of 1 g/day) or a matched placebo for a period of 8 weeks.ResultsOne hundred and seventeen subjects were assigned to either curcumin (n = 59) or placebo (n = 58) groups. Within-group analysis revealed significant reductions in serum concentrations of TNF-α, IL-6, TGF-β and MCP-1 following curcumin supplementation (p < 0.001). In the placebo group, serum levels of TGF-β were decreased (p = 0.003) but those of IL-6 (p = 0.735), TNF-α (p = 0.138) and MCP-1 (p = 0.832) remained unaltered by the end of study. Between-group comparison suggested significantly greater reductions in serum concentrations of TNF-α, IL-6, TGF-β and MCP-1 in the curcumin versus placebo group (p < 0.001). Apart from IL-6, changes in other parameters remained statistically significant after adjustment for potential confounders including changes in serum lipids and glucose levels, and baseline serum concentration of the cytokines.ConclusionResults of the present study suggest that curcumin supplementation significantly decreases serum concentrations of pro-inflammatory cytokines in subjects with MetS.     

Heart-type fatty acid binding protein is an early marker of myocardial damage after radiofrequency catheter ablation

ObjectivesRadiofrequency (RF) ablation of arrhythmias induces myocardial damage and release of biomarkers. This study aimed to assess the kinetics of heart-type fatty acid-binding protein (h-FABP), a cytosolic protein released after myocardial injury incurred by both atrial and ventricular RF ablation, compared to other markers of myocardial injury.Design and methodsh-FABP, cTnI, CK-MB_mass and myoglobin were evaluated in 30 patients with atrial or ventricular tachyarrhythmias before, immediately after and at 3, 6 and 24 h after the procedure.Resultsh-FABP increased immediately after the procedure in all subjects (6.6 ± 1.2 μg/L vs 2.7 ± 0.3, p < 0.001) but increased significantly only in ventricular ablations. The peak of h-FABP significantly correlates with the values of time for mean power of RF application in both the entire patient cohort and in ventricular ablations.Conclusionsh-FABP may be an early parameter for monitoring RF-induced lesions and the site of ablation was relevant for biomarker increase.