TP53 rs1042522 and rs8064946 variants in myocardial infarction.

OBJECTIVE: This study investigated the hypothesis that the single nucleotide polymorphisms (SNPs) of TP53 gene are related to a risk of myocardial infarction. METHODS: The coding SNP at codon 72 (rs1042522) and non-coding rs8064946 SNP were genotyped by polymerase chain reaction with sequence specific primers in 205 Czech patients with myocardial infarction and 148 Czech control subjects. RESULTS: The distribution of both SNPs was in agreement with the Hardy-Weinberg equilibrium and was similar to other European populations. Our power analysis showed 96 % of probability to detect an odd ratio equal to 2. Neither rs1042522 nor rs8064946 were associated with the risk of myocardial infarction. The haplotypes combined of rs1042522 and rs8064946 were not associated with myocardial infarction in the present study. CONCLUSION: The TP53 SNPs are not strongly associated with genetic predisposition to myocardial infarction (Tab. 3, Fig. 3, Ref. 23).

ANTIPHOSPHOLIPID SYNDROME AND MONOCYTES: NEW ASPECTS.

After discovery of antiphospholipid syndrome (APS) our understanding of molecular mechanisms of living matter has become more sophisticated and on this way monocytes has become crucial player, particularly in pathogenesis of APS. Thrombotic and non-thrombotic complications of APS could be explained by monocytes' activation too. But mechanisms underlying their activation are poorly investigated. So we aimed to determine transcriptional activity of monocytes after exposing them to low concentrations of lipopolysaccharide (LPS) and LPS+ATP using comparative of RT-PCR. Our study included eleven women suffering from recurrent miscarriages and APS (mean age 30±5,6 years). Nine healthy women (mean age of 29±8,5 years) without a positive family history of APS, autoimmune diseases and thrombosis were chosen as a control group. The results showed increasing levels of TLR2, IL-23, CCL2, CXCL10, IL-1ß and IL-6 in APS cells, while in healthy cells LPS resulted in IL-6 and STAT3 elevated mRNAs. Double stimulation of APS cells resulted in decreased mRNA levels of CCL-2, IL-1ß, and mRNA NLRP3 in healthy cells. At the same time TLR2 mRNAs were elevated in both groups after double stimulation. Thus increased sensitivity of APS cells to LPS may contribute to thrombus formation. Low concentration of ATP diminishes LPS-induced inflammatory state of APS monocytes, which might be one of potential regulatory mechanisms.

Grids from bands, or bands from grids? An examination of the effects of single unit contamination on grid cell firing fields.

Neural recording technology is improving rapidly, allowing for the detection of spikes from hundreds of cells simultaneously. The limiting step in multielectrode electrophysiology continues to be single cell isolation. However, this step is crucial to the interpretation of data from putative single neurons. We present here, in simulation, an illustration of possibly erroneous conclusions that may be reached when poorly isolated single cell data are analyzed. Grid cells are neurons recorded in rodents, and bats, that spike in equally spaced locations in a hexagonal pattern. One theory states that grid firing patterns arise from a combination of band firing patterns. However, we show here that summing the grid firing patterns of two poorly resolved neurons can result in spurious band-like patterns. Thus, evidence of neurons spiking in band patterns must undergo extreme scrutiny before it is accepted. Toward this aim, we discuss single cell isolation methods and metrics.

Transcriptional activity of neutrophils exposed to high doses of colchicine: short communication.

Colchicine is an antimitotic drug which binds to tubulin and at high doses results in cytoskeleton disruption. Colchicine is believed to be an anti-inflammatory agent, though its modulatory effects on the level and transcriptional activity of genes is still a matter of debate. There is growing evidence that alterations in the cytoskeleton exert specific effects on the expression of various genes. This study was undertaken to analyze whether disrupting the microtubule cytoskeleton by colchicine modulates transcriptional levels of MEFV, NF-κB p65, NLRP3, HMGB1, and caspase-3 in neutrophils from patients with familial Mediterranean fever (FMF) and healthy subjects. In the present study, colchicine caused increased expression of NLRP3 (p=0.007) and MEFV (p=0.03), but had no effect on caspase-3, NF-κB p65 and HMGB1 genes in healthy neutrophils. FMF neutrophils were less responsive to the drug treatment. This study supports the hypothesis that, being an anti-inflammatory agent, colchicine at relatively high concentrations might lead to the activation of pro-inflammatory signalling pathways in neutrophils.

Closed-loop optical neural stimulation based on a 32-channel low-noise recording system with online spike sorting.

OBJECTIVE: Closed-loop operation of neuro-electronic systems is desirable for both scientific and clinical (neuroprosthesis) applications. Integrating optical stimulation with recording capability further enhances the selectivity of neural stimulation. We have developed a system enabling the local delivery of optical stimuli and the simultaneous electrical measuring of the neural activities in a closed-loop approach. APPROACH: The signal analysis is performed online through the implementation of a template matching algorithm. The system performance is demonstrated with the recorded data and in awake rats. MAIN RESULTS: Specifically, the neural activities are simultaneously recorded, detected, classified online (through spike sorting) from 32 channels, and used to trigger a light emitting diode light source using generated TTL signals. SIGNIFICANCE: A total processing time of 8 ms is achieved, suitable for optogenetic studies of brain mechanisms online.

Colchicine modulates expression of pro-inflammatory genes in neutrophils from patients with familial Mediterranean fever and healthy subjects.

Colchicine (Col) is a microtubule depolymerizing drug, widely used for treatment of familial Mediterranean fever (FMF). Mechanisms by which Col exerts its beneficial effects are not yet completely understood, especially with respect to gene expression in polymorphonuclear neutrophils (PMNs), the main effector cells in acute inflammatory attacks of FMF. This study was, therefore, designed to elucidate possible modulatory effect of Col on expression of inflammation-related genes in circulating PMNs from 16 FMF patients in the remission period and 11 healthy subjects. In vitro effect of Col exposure (1 microg/ml) on expression of 8 selected genes was examined using quantitative real-time RT-PCR. Col up-regulated expression of IL-8 and IL-1beta genes in FMF (13-fold and 2.7-fold, p less than 0.05, respectively) and healthy (3-fold and 6.5-fold, p less than 0.05, respectively) PMNs, and down-regulated caspase-1 in FMF neutrophils (3-fold, p less than 0.05). In FMF PMNs treated with Col mRNAs of IL-8 (51-fold, p less than 0.01) and c-FOS (7-fold, p less than 0.05) transcripts were elevated compared to those from healthy subjects. By contrast, caspase-1 mRNA was decreased in FMF neutrophils compared to healthy cells (1.6-fold, p less than 0.05). Hereby, we provide evidence that, at least in vitro, Col displays pro-inflammatory potential in respect to IL-1beta and IL-8 genes. At the same time, our findings implicate suppression of caspase-1 expression by Col as a potential mechanism for its effects in FMF treatment.

MBL2 gene variation affecting serum MBL is associated with prosthetic joint infection in Czech patients after total joint arthroplasty.

Prosthetic joint infection (PJI) is a serious complication of the total joint arthroplasty (TJA). Serum mannose-binding lectin (MBL), a pattern recognition receptor, is involved in antibacterial immune response. This study investigated whether functional variants of the MBL2 gene may be associated with the risk of PJI. MBL2 -550 (H/L, rs11003125), MBL2 -221 (Y/X, rs7096206) and MBL2 +54 (G/A, rs1800450) single nucleotide polymorphisms (SNP) were genotyped in 112 PJI patients and two control groups: 245 patients with aseptic TJA and 196 Czech population controls without TJA. Serum MBL concentration was assessed in PJI patients (n = 92) and aseptic TJA controls (n = 56). The distribution of MBL2 genotypes complied with the Hardy-Weinberg equilibrium in all investigated groups. Importantly, MBL2 -550 L allele (allelic frequency, 0.72) and LL genotype (genotype frequency, 0.51) were more frequent among PJI patients compared to aseptic TJA controls (L allele: 0.63, P = 0.016, P(c) = 0.048; LL genotype: 0.39, P = 0.037, P(c) > 0.05) and to Czech population controls (L allele: 0.61, P = 0.010, P(c) = 0.030; LL genotype: 0.35, P = 0.006, P(c) = 0.018), respectively. Regarding MBL protein, the MBL2 -550 L carriers presented with lower serum MBL concentrations than non-carriers (median; 593 vs 1876 ng/ml; P < 0.01). Similarly, the carriage of MBL2 -221 X and 54 A alleles was associated with lower serum MBL concentrations (P < 0.01). In conclusion, MBL2 -550 genetic variant(s) associated with low serum concentration of MBL protein can increase the risk of PJI.

Cryopreserved and lyophilized cultured epidermal allografts in the treatment of leg ulcers: a pilot study.

BACKGROUND: In the conservative therapy of venous leg ulcers modern types of dressings are used most frequently. In the past 20 years 'active wound dressings' - cultured epidermal keratinocytes as autografts and allografts - were also introduced in the management of leg ulcers. METHODS: The aim of our study was to compare the effect of cryopreserved and lyophilized cultured epidermal allografts in the treatment of venous leg ulcers. Evaluation of the therapy was documented as photodocumentation, planimetry, healing time and evaluation of pain relief over a 3-month period after application. Fifty patients with venous leg ulcers were selected. Twenty-five patients (group I) were treated with cryopreserved keratinocytes and 25 (group II) with lyophilized keratinocytes. RESULTS: The final evaluation 3 months after the application of allografts showed 84% of healed ulcers in group I and 80% in group II. The number of healed ulcers and the healing rate both showed no statistically significant differences. The size of the ulcer was reduced by half during the first week in both groups. The size differences during the first week are statistically significant in both groups and they are comparable (P < 0.001). The intensity of the pain was statistically significantly reduced during the first week after application in both groups (P < 0.001). CONCLUSIONS: The cryopreserved and lyophilized cultured allografts are comparable in healing rate, course of healing and relief of pain, and also in planimetric changes during the healing of venous leg ulcers. Lyophilized allografts are more convenient for routine use than cryopreserved allografts as they can be stored at room temperature. These results could give rise to the more frequent use of lyophilized allografts in slow-healing venous leg ulcers.

Successful treatment of chronic venous leg ulcers with lyophilized cultured epidermal allografts.

The case of a 67-year-old woman with chronic venous leg ulcers and severe gonarthrosis is described. In spite of intensive therapy, the leg ulcers had persisted for 4 years and made the intended orthopaedic operation of the right knee impossible. The patient was treated with lyophilized cultured epidermal allografts and her leg ulcers healed within 40 days. Lyophilized cultured epidermal allografts represent a modern type of active wound dressing that leads to rapid healing of chronic venous leg ulcers and enables patients to undergo surgical intervention.

Determination of gold using clay modified carbon paste electrode.

Sorption of gold(III) chlorocomplexes was studied by means of a carbon paste electrode modified with montmorillonite. Anionic exchange behavior was found in chloride media with low ionic strength. Anionic sorption of [AuCl4]- can be used as a preconcentration step to the determination of Au(III). Linear calibration dependences were found in the concentration range 4.06 x 10(-6) - 1.22 x 10(-5) mol/L Au(III) after 5 min of sorption and in the range 8.12 x 10(-7) - 6. 1 x 10(-6) mol/L after 10 min of sorption. Interferences of several anions and cations were studied. Model samples of table water were analyzed.

Voltammetric copper(II) determination with a montmorillonite-modified carbon paste electrode.

The clay mineral montmorillonite has been tested as modifier for the carbon paste electrode with a novel electrode modification technique. The differential pulse voltammetric determination of copper(II) by means of this modified carbon paste electrode has been studied. A detection limit of 4x10(-8) mol/l has been achieved after 10 min preconcentration under open circuit conditions with subsequent anodic stripping voltammetry. The calibration curve for Cu(II) is linear in the range of 4x10(-8)-8x10(-7) mol/l. Pb interferes in a 10-fold molar and Cd and Hg in a 100-fold molar excess. The interference by humic ligands is significant.