BBA clinical最新文献

Background

Oxidative stress occurs in many neurodegenerative diseases including Alzheimer's disease (AD) and evidence suggests that specific proteins are oxidised in individual diseases. Thus measures of oxidised proteins such as in human biological samples could represent potential disease-specific biomarkers. Protein carbonylation is considered to be an important marker of oxidative stress. In AD in particular, the peptidyl prolyl isomerase, Pin1, has been shown to be sensitive to metal-catalysed oxidation with the addition of carbonyl side-chains.

Methods

Based on this protein modification we developed a novel, enzyme-linked sandwich immunoassay for the quantification of oxidised Pin1 (oxPin1) in human brain tissue samples.

Results

We successfully developed an ELISA for the measurement of oxidised Pin1 in biological samples and measured oxPin1 in hippocampal tissue extracts from controls and AD, which showed an increased ratio of oxPin1 to total Pin1 in patients with early AD pathology compared with controls.

Conclusions

We show that oxidised proteins, in this case oxPin1, can be measured using the developed ELISA. In addition, our results support the presence of increased oxidative stress in the early stages of AD pathology and show that the oxPin1/Pin1 ratio could indicate early stage pathology. This warrants further investigation in other biological fluids.

General significance

Importantly, further development and adaption of the assay design will enable multi-functional use for the quantification of oxidised proteins in tissues and biological fluids that may be used in investigating the role of oxidised proteins in a range of neurodegenerative diseases, particularly in which disease-specific protein oxidation has been implicated.

Intravesical Bacillus Calmette–Guerin (BCG) vaccine is the preferred first line treatment for non-muscle invasive bladder carcinoma (NMIBC) in order to prevent recurrence and progression of cancer. There is ongoing need for the rational selection of i) BCG dose, ii) frequency of BCG administration along with iii) synergistic adjuvant therapy and iv) a reliable set of biochemical markers relevant to tumor response. In this review we evaluate cellular and molecular markers pertinent to the immunological response triggered by the BCG instillation and respective mathematical models of the treatment. Specific examples of markers include diverse immune cells, genetic polymorphisms, miRNAs, epigenetics, immunohistochemistry and molecular biology ‘beacons’ as exemplified by cell surface proteins, cytokines, signaling proteins and enzymes. We identified tumor associated macrophages (TAMs), human leukocyte antigen (HLA) class I, a combination of Ki-67/CK20, IL-2, IL-8 and IL-6/IL-10 ratio as the most promising markers for both pre-BCG and post-BCG treatment suitable for the simulation studies.

The intricate and patient-specific nature of these data warrants the use of powerful multi-parametral mathematical methods in combination with molecular/cellular biology insight and clinical input.

Background

Acute myeloid leukemia (AML) patients with highly active AKT tend to do poorly. Cell cycle arrest and apoptosis are tightly regulated by AKT via phosphorylation of GSK3α and β isoforms which inactivates these kinases. In the current study we examine the prognostic role of AKT mediated GSK3 phosphorylation in AML.

Methods

We analyzed GSK3α/β phosphorylation by reverse phase protein analysis (RPPA) in a cohort of 511 acute myeloid leukemia (AML) patients. Levels of phosphorylated GSK3 were correlated with patient characteristics including survival and with expression of other proteins important in AML cell survival.

Results

High levels of p-GSK3α/β correlated with adverse overall survival and a lower incidence of complete remission duration in patients with intermediate cytogenetics, but not in those with unfavorable cytogenetics. Intermediate cytogenetic patients with FLT3 mutation also fared better respectively when p-GSK3α/β levels were lower. Phosphorylated GSK3α/β expression was compared and contrasted with that of 229 related cell cycle arrest and/or apoptosis proteins. Consistent with p-GSK3α/β as an indicator of AKT activation, RPPA revealed that p-GSK3α/β positively correlated with phosphorylation of AKT, BAD, and P70S6K, and negatively correlated with β-catenin and FOXO3A. PKCδ also positively correlated with p-GSK3α/β expression, suggesting crosstalk between the AKT and PKC signaling pathways in AML cells.

Conclusions

These findings suggest that AKT-mediated phosphorylation of GSK3α/β may be beneficial to AML cell survival, and hence detrimental to the overall survival of AML patients. Intrinsically, p-GSK3α/β may serve as an important adverse prognostic factor for a subset of AML patients.

Background

Determination of the role of steroid hormones in expression and regulation of endometrial glucose transport 4 (GLUT4) in humans is important for understanding endometrial disorders such as polycystic ovary syndrome (PCOS), a common hormone-imbalance disease.

Methods

Endometrial biopsy samples were collected from non-PCOS patients with regular menstrual cycles or with hyperplasia and from PCOS patients with or without hyperplasia. In addition, endometrial tissues from postmenopausal women were incubated with human chorionic gonadotropin (hCG, 10 IU/ml), 17β-estradiol (E2, 10 nM), progesterone (P4, 100 nM), or a combination of E2 and P4 for 24 h. The expression of GLUT4 was measured at the mRNA level using quantitative real-time polymerase chain reaction (qRT-PCR) and at the protein level using Western blot analysis and immunohistochemistry.

Results

A cyclical change in GLUT4 expression pattern was observed in non-PCOS patients, and a high level of GLUT4 expression was seen in the proliferative phase compared to the secretory phase. Low levels of GLUT4 expression were found in PCOS patients compared to menstrual cycle phase-matched non-PCOS patients, and there was no significant change in GLUT4 expression in PCOS patients during the menstrual cycle. GLUT4 was localized in both epithelial and stromal cells, with notable changes in epithelial cells. We postulate that decreased GLUT4 expression might be regulated by steroid hormones. In support of this, we showed that in cultured endometrial tissues hCG and E2 alone had no effect on GLUT4 expression. However, P4 alone and P4 in combination with E2 decreased GLUT4 expression. Compared with non-PCOS controls, PCOS patients with endometrial hyperplasia exhibited decreased GLUT4 expression in particular in the epithelial cells.

Conclusion

We conclude that P4 can induce changes in endometrial GLUT4 expression during the menstrual cycle and that abnormal hormonal conditions such as PCOS disrupt normal patterns of GLUT4 expression in endometrial cells.

Background

Despite many recent advances in endovascular therapy (EVT), peripheral artery disease (PAD) is an increasing health problem with high mortality. Heart-type fatty acid-binding protein (H-FABP) and high-sensitivity troponin T (hsTnT) are markers of ongoing myocardial damage and have been reported to be useful indicators of future cardiovascular events. However, it remains to be determined whether H-FABP and hsTnT can predict adverse clinical outcomes in patients with PAD.

Methods and results

We enrolled 208 de novo PAD patients who underwent EVT. Serum H-FABP and hsTnT were measured in all patients before EVT. During the median follow-up period of 694 days, there were 40 major adverse cardiovascular and cerebrovascular events (MACCEs) including all-cause deaths, and re-hospitalizations due to cardiovascular and cerebrovascular diseases and amputations. H-FABP and hsTnT were found to be higher in patients with critical limb ischemia (CLI) compared to those without this condition. Multivariate Cox proportional hazard regression analysis revealed that both H-FABP and hsTnT were independent predictors of MACCEs after adjustment for confounding factors. Kaplan–Meier analysis demonstrated that patients in the highest tertile according to H-FABP levels, as well as those in the highest hsTnT tertile, were at greatest risk for MACCEs. The net reclassification index was significantly improved by the addition of H-FABP as well as the addition of hsTnT to traditional risk factors.

Conclusion

The myocardial damage markers H-FABP and hsTnT were increased in PAD patients with CLI and could predict MACCEs in PAD patients.

Objectives

Traditional markers of cerebral small vessel disease (SVD) are related to cognition and cognitive decline, but this relation is weak. Therefore other factors may determine the transition from intact cognitive performance to cognitive decline, such as the damage of the cerebral white matter at the microstructural level. Little is known about the association between microstructural integrity of the white matter and changes in cognition. In this study we investigated the relation between baseline microstructural integrity and change in cognitive function.

Methods

503 participants of the RUN DMC study with SVD without dementia, 398 of whom (79.1%) underwent repeated cognitive testing at follow-up, with a mean follow-up time of 5.4 years (± SD 0.2), and among others FLAIR MRI and diffusion tensor imaging (DTI). At baseline Mean Diffusivity (MD) and mean Fractional Anisotropy (FA) were measured in both white matter hyperintensities (WMH) and normal appearing white matter (NAWM). A linear regression analysis was performed assessing the association between baseline diffusion parameters and decline in cognitive domains.

Results

An inverse association was found between baseline MD in the NAWM and decline in Cognitive Index (β = 0.17; p = 0.035), adjusted for age, sex, education, presence of depressive symptoms at baseline, normalized TBV, lacunes and WMH volume. However, no significant associations were found between diffusion parameters and decline in any cognitive domain after Bonferroni correction.

Conclusions

In contrast to cross-sectional studies, in older adults with SVD microstructural integrity of the white matter as assessed with DTI is not related to decline in global cognitive function or any other subdomain.

Background

Long chain polyunsaturated fatty acids (LCPUFAs) are biologically active fatty acids which regulate placental angiogenesis, inflammation, and oxidative stress. Abnormalities in these aspects have been associated with preeclampsia (PE). Further, placenta has a heterogeneous structure with differential vascularization across different regions. We therefore hypothesize that the distribution of fatty acids in various regions of the placenta is altered in PE leading to poor fetal outcome.

Methods

In this cross-sectional study we recruited 69 normotensive control (NC) and 44 women with PE. PE women were further classified as those delivered preterm (PTPE, n = 24) and at term (TPE, n = 20). Fatty acid levels were analyzed from placental samples from four different regions (CF—central fetal, PF—peripheral fetal, CM—central maternal and PM—peripheral maternal).

Results

In the NC placenta, AA levels were lower (p < 0.05) in CM as compared with CF region. However, such differences were not seen in the TPE and PTPE. In contrast, the DHA levels varied between regions only in the PTPE placenta. Between groups, DHA levels were lower (p < 0.05 for both) in the CM and CF regions of the PTPE as compared with NC. The levels of DHA in TPE placenta were similar to NC. AA levels were lower (p < 0.05 for both) in CF region of TPE and PF region of PTPE placenta than NC.

Conclusions

There is differential pattern of LCPUFA distribution across various regions of the NC, TPE and PTPE placenta. This may have implications for placental growth and development as well as transfer of LCPUFA to the fetus.

Backgrounds

Glycero-lysophospholipids (glycero-LPLs), which are known to exert potent biological activities, have been demonstrated to be secreted from activated platelets in vitro; however, their association with platelet activation in vivo has not been yet elucidated. In this study, we investigated the correlations between the blood levels of each glycero-LPL and serotonin, a biomarker of platelet activation, in human subjects to elucidate the involvement of platelet activation in glycero-LPLs in vivo.

Methods and Results

We measured the plasma serotonin levels in 141 consecutive patients undergoing coronary angiography (acute coronary syndrome, n = 38; stable angina pectoris, n = 71; angiographically normal coronary arteries, n = 32) and investigated the correlations between the plasma levels of serotonin and glycero-LPLs. The results revealed the existence of a specific and significant association between the plasma serotonin and plasma lysophosphatidylserine (LysoPS) levels. On the contrary, regular aspirin intake failed to affect the plasma LysoPS levels despite the fact that the plasma lysophosphatidic acid, lysophosphatidylethanolamine, lysophosphatidylglycerol, and lysophosphatidylinositol levels were lower in those who had taken aspirin regularly.

Conclusion

We found a specific positive correlation between the blood levels of serotonin and LysoPS, a new lipid mediator. Thus, LysoPS might be specifically involved in strong platelet activation, which is associated with the release of serotonin.

General Significance

Our present results suggest the possible involvement of LysoPS in the pathogenesis of atherosclerotic diseases.

Background

The role of pattern of circulating endothelial cell-, platelet-, and monocyte-derived microparticles in metabolic syndrome (MetS) patients with chronic heart failure (CHF) is not still understood.

The aim of the study was to investigate a pattern of circulating MPs in MetS patients with CHF in relation to neurohumoral and inflammatory activation.

Methods

The study retrospectively involved 101 patients with MetS (54 subjects with CHF and 47 patients without CHF) without documented coronary artery stenosis > 50% at least of one artery and 35 healthy volunteers. Biomarkers were measured at baseline of the study. Circulating MPs were phenotyped by flow cytometry technique.

Results

The results of the study have shown that numerous of the circulating platelet-derived and monocyte-derived MPs in subjects with MetS (with or without CHF) were insufficiently distinguished from the level obtained in healthy volunteers. We found an elevated level of CD31 +/annexin V + MPs in association with a lower level of CD62E + MPs. All these led to decreased CD62E + to CD31 +/annexin V + ratio among patients with MetS in comparison with healthy volunteers, as well as in MetS patients with CHF compared with those who did not demonstrated CHF. Therefore, we found that biomarkers of biomechanical stress (NT-proBNP) and inflammation (hs-CRP, osteoprotegerin) remain statistically significant predictors for decreased CD62E + to CD31 +/annexin V + ratio in MetS patients with CHF.

In conclusion, decreased CD62E + to CD31 +/annexin V + ratio reflected impaired immune phenotype of MPs may be discuss surrogate marker of CHF development in MetS population.

Background

Stroke prevention by warfarin, a vitamin K antagonist, has been an integral part in the management of atrial fibrillation. Vitamin K-dependent matrix Gla protein (MGP) has been known as a potent inhibitor of arterial calcification and osteoporosis. Therefore, we hypothesized that warfarin therapy affects bone mineral metabolism, vascular calcification, and vascular endothelial dysfunction.

Methods

We studied 42 atrial fibrillation patients at high-risk for atherosclerosis having one or more coronary risk factors. Twenty-four patients had been treated with warfarin for at least 12 months (WF group), and 18 patients without warfarin (non-WF group). Bone alkaline phosphatase (BAP) and under carboxylated osteocalcin (ucOC) and receptor activator of nuclear factor-kappa B ligand (RANKL) were measured as bone metabolism markers. Reactive hyperemia-peripheral arterial tonometry (RH-PAT) index measured by Endo-PAT2000 was used as an indicator of vascular endothelial function.

Results

There were no significant differences in patient background characteristics and other clinical indicators between the two groups. In WF group, the ucOC levels were significantly higher than those in the non-WF group (10.3 ± 0.8 vs. 3.4 ± 0.9 ng/mL; P < 0.01), similarly, the RANKL levels in the WF group were higher than those in the non-WF group (0.60 ± 0.06 vs. 0.37 ± 0.05 ng/mL; P = 0.007). Moreover, RH-PAT index was significantly lower in the WF group compared to those in the non-WF group (1.48 ± 0.11 vs. 1.88 ± 0.12; P = 0.017).

Conclusions

Long-term warfarin therapy may be associated with bone mineral loss and vascular calcification in 60–80 year old hypertensive patients.