Usefulness of material recovered from distal embolic protection devices after carotid angioplasty for proteomic studies.

PURPOSE: To demonstrate the usefulness of biologic material obtained from distal embolic protection devices (DEPDs) used in carotid angioplasty for the study of atherosclerosis protein markers and to establish the effect of systemic inflammation on the protein expression of carotid atheromatous plaques. MATERIALS AND METHODS: Two-dimensional gel electrophoresis and mass spectrometry were used to study proteins obtained from debris captured in DEPDs from patients who underwent carotid angioplasty. In addition, protein expression obtained from angioplasty samples in patients with different types of systemic inflammation (measured by serum levels of high-sensitivity C-reactive protein [CRP] with a cutoff value of 3 mg/L) was compared. Finally, immunohistochemistry of atherosclerotic plaques obtained by endarterectomy was used to validate the results obtained using DEPDs. RESULTS: Proteomic studies were successfully performed using debris from DEPDs. Protein expression differences were found in debris from patients with high systemic inflammation compared with debris from patients with low systemic inflammation. Annexin A5 (ANXA5), haptoglobin precursor, purine nucleoside phosphorylase, transgelin-2 (TAGLN2), and bisphosphoglycerate mutase were upregulated in debris from patients with high systemic inflammation, and proteasome subunit 8 beta type and glutathione-S-transferase kappa 1 (GSTK1) levels were higher in debris from patients with low levels of systemic inflammation. CONCLUSIONS: Atherosclerotic plaque debris captured in DEPDs is a suitable and valid source of material for proteomic studies of atherosclerosis. Protein expression in DEPD debris is affected by systemic inflammation.

Evaluation of an O2-Substituted (1-3)-ß-D-Glucan, Produced by Pediococcus parvulus 2.6, in ex vivo Models of Crohn's Disease.

1,3-ß-glucans are extracellular polysaccharides synthesized by microorganisms and plants, with therapeutic potential. Among them, the O2-substituted-(1-3)-ß-D-glucan, synthesized by some lactic acid bacteria (LAB), has a prebiotic effect on probiotic strains, an immunomodulatory effect on monocyte-derived macrophages, and potentiates the ability of the producer strain to adhere to Caco-2 cells differentiated to enterocytes. In this work, the O2-substituted-(1-3)-ß-D-glucan polymers produced by GTF glycoyltransferase in the natural host Pediococcus parvulus 2.6 and in the recombinant strain Lactococcus lactis NZ9000[pNGTF] were tested. Their immunomodulatory activity was investigated in an ex vivo model using human biopsies from patients affected by Crohn's disease (CD). Both polymers had an anti-inflammatory effect including, a reduction of Interleukine 8 both at the level of its gene expression and its secreted levels. The overall data indicate that the O2-substituted-(1-3)-ß-D-glucan have a potential role in ameliorating inflammation via the gut immune system cell modulation.

Temporal profile and clinical significance of serum neuron-specific enolase and S100 in ischemic and hemorrhagic stroke.

BACKGROUND: Neuron-specific enolase (NSE) and S100 protein are implicated in several brain injuries, including stroke. Our objective was to analyze the temporal profile and the clinical significance of NSE and S-100 in acute ischemic (IS) and intracerebral hemorrhage (ICH). METHODS: We studied 224 patients with IS and 44 patients with ICH. Computerized tomography (CT) scans were performed to assess infarct volume. Stroke severity was evaluated using the National Institute of Health Stroke Scale (NIHSS), and functional outcome at 3 months with the modified Rankin Scale (mRS). Serum NSE and S100 protein were measured using an electrochemiluminescence-immunoassay. RESULTS: Peak values were found at 72 h for NSE and at 24 h for S100 in IS. For ICH, peak values were found at 24 h for both NSE and S100. At these time intervals S100 and NSE correlated with the NIHSS score and were independently associated with poor outcome. CONCLUSIONS: High serum NSE and S100 are associated with poor outcome in IS, and high serum NSE is associated with poor outcome in ICH. These findings suggest the potential utility of NSE and S100 as prognostic markers for acute stroke.

Increased body iron stores are associated with poor outcome after thrombolytic treatment in acute stroke.

BACKGROUND AND PURPOSE: Iron overload has been associated with greater oxidative stress and brain injury in experimental cerebral ischemia and reperfusion. This study investigates whether high serum ferritin levels, as an index of increased cellular iron stores, are associated with poor outcome, hemorrhagic transformation, and brain edema after treatment with tissue plasminogen activator in patients with acute ischemic stroke. METHODS: A total of 134 consecutive patients treated with intravenous tissue plasminogen activator were prospectively studied in four centers. Serum ferritin levels were determined at baseline, 24 and 72 hours after treatment. Cranial computed tomography was performed on admission and at 24 to 36 hours after tissue plasminogen activator infusion. Stroke severity and outcome were evaluated by using the National Institute of Health Stroke Scale and the modified Rankin Scale. RESULTS: Computed tomography showed hemorrhagic transformation in 27 patients (hemorrhagic infarction in 15 and parenchymal hematoma in 12; symptomatic in four) and brain swelling with midline shift in 15. Poor outcome (modified Rankin Scale >2) at 90 days was observed in 54.5% of patients. Ferritin levels at baseline were higher in patients with poor outcome at 90 days (median [quartiles], 165 [98,307] versus 17 [12,37] ng/mL; P<0.001) and in those who developed parenchymal hematoma (P=0.006), symptomatic hemorrhagic transformation (P=0.008), and severe brain edema (P<0.001). Serum ferritin levels higher than 79 ng/mL before tissue plasminogen activator treatment were independently associated with poor outcome (OR, 117 [95% CI, 25 to 557]). CONCLUSIONS: Increased body iron stores are associated with poor outcome, symptomatic hemorrhagic transformation, and severe edema in patients treated with tissue plasminogen activator after ischemic stroke. These findings suggest that iron overload may offset the beneficial effect of thrombolytic therapies.

Proteomic analysis of 1α,25-dihydroxyvitamin D3 action on human colon cancer cells reveals a link to splicing regulation.

1α,25-Dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) and other vitamin D compounds are promising molecules in the prevention and therapy of colon cancer and other neoplasias. To study the mechanism of action of 1,25(OH)(2)D(3) in colon cancer cells, we carried out a comparative proteomic analysis of the nuclear fractions of SW480-ADH cells treated with 1,25(OH)(2)D(3) or vehicle during 8 or 48h. 2D-DIGE analysis combined with MALDI-TOF-TOF mass spectrometry interrogation led to the identification of 59 differentially expressed unique proteins. Most identified proteins were nuclear, but several cytoskeleton-associated proteins were also detected. A good concordance between changes in expression at protein and RNA levels was observed for the validated proteins. A large group of identified proteins, such as SFPQ, SMARCE, KHSRP, TARDBP and PARP1, were involved in RNA processing or modification and have been ascribed to the spliceosome compartment of human cells. In addition, a smaller group of proteins (ERM (Ezrin, Radixin, Moesin) family, VCL, CORO1C, ACTB) were cytoskeleton-associated and played a role in cell adhesion and morphology. These results confirm the induction of epithelial phenotype by 1,25(OH)(2)D(3) and suggest a role for vitamin D compounds in the regulation of the spliceosome and thus, in alternative splicing and possibly microRNA synthesis in colon cancer cells.

Relative efficiency of the linkage disequilibrium mapping approach in detecting candidate genes for schizophrenia in different European populations.

Detection of susceptibility genes in indirect association studies depends not only on the degree of linkage disequilibrium between the disease variant and the SNP marker but also on the difference in their allele frequencies. Little is known about how variations in these parameters may affect the power of indirect association studies among related populations. Toward this goal, we genotyped 40 SNPs at four loci in samples from three European populations, Galician, Greek, and Norwegian. We compared the relative efficiency of all pairs of SNPs in detecting each other in each one of the populations. Our results show that a low percentage of marker SNPs may detect association in some populations but be totally ineffective in others. Therefore, these differences have to be an additional factor to consider when a replication study fails to confirm initial associations, especially if the replication is focused on very few markers.