The prevalence of Chlamydia pneumoniae in the aortic wall and in peripheral blood of patients scheduled for coronary artery bypass grafting.

Some reports confirm a potential role of Chlamydia pneumoniae (ChP) in atherogenesis. In order to explore possible association between ChP and atherosclerosis, investigations were carried out in which the frequency of ChP in the arterial wall and peripheral blood was assessed in a group of patients with chronic coronary artery disease (CAD). Fifty-seven patients were enrolled in the study, 13 women and 44 men aged 61.8±6.5 (47-74), with previously diagnosed CAD, scheduled for planned coronary artery bypass grafting due to clinical indications. Vessel specimens retrieved from the ascending aorta (as a part of routine proximal venous graft development procedure) and peripheral blood mononuclear cells (PBMCs) from venous blood were evaluated for the presence of ChP DNA. Genomic DNA was extracted from PBMCs and vessel specimens. Quantitative real-time polymerase chain reaction (qPCR) was performed to detect ChP DNA. A statistically more frequent occurrence of ChP was observed in aortic tissues compared to blood samples (70.2% vs 56.1%, respectively). Similarly, the number of ChP DNA genomic copies [n/1µg genomic DNA] was significantly higher in tissue specimens compared to blood samples (89±91 vs 41±77, respectively; p=0.0046). In patients without ChP in blood specimens, we observed significantly higher amounts of ChP in tissue specimens compared to patients with ChP in blood specimens (156±71 vs 107±88, respectively; p=0.0453). No correlation was found between the number of ChP DNA copies [n/1µg genomic DNA] in blood and in aortic specimens. The infection of ChP in the aortic wall was connected with hypercholesterolemia (p=0.029) and diabetes (p=0.03). We conclude that Chlamydia pneumoniae is a pathogen frequently occurring in the aortic wall of patients with CAD. The occurrence of ChP DNA in the aortic tissue is related to classic CAD risk factors such as diabetes and dyslipidemia.

The Correlation of PPARα Activity and Cardiomyocyte Metabolism and Structure in Idiopathic Dilated Cardiomyopathy during Heart Failure Progression.

This study aimed to define relationship between PPARα expression and metabolic-structural characteristics during HF progression in hearts with DCM phenotype. Tissue endomyocardial biopsy samples divided into three groups according to LVEF ((I) 45-50%, n = 10; (II) 30-40%, n = 15; (III) <30%, n = 15; and control (donor hearts, >60%, n = 6)) were investigated. The PPARα mRNA expression in the failing hearts was low in Group (I), high in Group (II), and comparable to that of the control in Group (III). There were analogous changes in the expression of FAT/CD36 and CPT-1 mRNA in contrast to continuous overexpression of GLUT-4 mRNA and significant increase of PDK-4 mRNA in Group (II). In addition, significant structural changes of cardiomyocytes with glycogen accumulation were accompanied by increased expression of PPARα. For the entire study population with HF levels of FAT/CD36 mRNA showed a strong tendency of negative correlation with LVEF. In conclusion, PPARα elevated levels may be a direct cause of adverse remodeling, both metabolic and structural. Thus, there is limited time window for therapy modulating cardiac metabolism and protecting cardiomyocyte structure in failing heart.

The uefa euro 2012 anti-doping programme - scientific review.

The final tournament of the UEFA European Football Championship is one of the top sporting events in the world, and a high-profile event of this kind requires a well-planned and well-executed anti-doping programme to ensure the integrity of results in the competition. UEFA EURO 2012 presented a unique logistical challenge, with the tournament spread across two countries, both covering a large geographical area. This paper discusses the planning and delivery of both the pre tournament out-of-competition (OOC) testing programme and the in-competition (IC) programme, as well as reviewing the activities of doping control officers (DCOs), the whereabouts programme and assessing the sample collection and transport process. The analytical approach applied is also discussed, along with an overview of the distribution of T/E ratios and blood parameters.

Identification and characterization of urinary prenylamine metabolites by means of liquid chromatography-tandem mass spectrometry.

Prenylamine is a vasodilator of phenylalkylamine structure and was used for the treatment of angina pectoris, until reports of undesirable effects including ventricular tachycardia led to a decreasing use of the drug in the 1980s. Metabolic N-dealkylation of orally ingested prenylamine can liberate amphetamine in humans and cause positive findings for amphetamine in doping and forensic analysis. In 2010, the World Anti-Doping Agency (WADA) classified prenylamine as a non-specified stimulant according to the 2010 Prohibited List, thus banning its use in sports in-competition. Supporting the development of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) based detection method, a post-administration urine sample following a single oral prenylamine ingestion (Segontin(®) 60 mg) was analyzed for urinary metabolites. The LC-separated analytes were ionized in positive electrospray ionization (ESI) mode and detected as protonated ions using an AB Sciex TripleTOF 5600 quadrupole-time-of-flight hybrid mass spectrometer. Over 40 phase I metabolites were detected, including previously unknown mono- bis-, tris- and tetra-hydroxylated prenylamine, several hydroxylated and methoxylated prenylamine metabolites and (hydroxylated) diphenylpropylamine. Investigation of the collision-induced dissociation behaviours of the metabolites by high resolution/high accuracy mass spectrometry allowed for the assignment of the nature and the site of observed metabolic transformations. The most abundant phase I metabolite was confirmed as p-hydroxy-prenlyamine by chemical synthesis and stable isotope labelling of reference material. An existing routine screening assay based on direct injection and LC-MS/MS analysis of urine was modified and validated according to common guidelines, in order to allow for the detection of p-hydroxy-prenylamine in sports drug testing. The assay demonstrated the ability to detect the target metabolite at 0.1 ng/ml at intra- and inter-day imprecisions below 10%.

Problems of the use of pseudoephedrine by athletes.

Pseudoephedrine (PSE) as a sympathomimetic is an ingredient of many proprietary medicines which are available on the medical market over the counter (OTC drugs). It can be converted to cathine (CATH, norpseudoephedrine) inside the body. Until the end of 2003, PSE had been a banned substance in sport in case its urinary concentration was greater than 25 mircog/ml. Then the World Anti-Doping Agency (WADA) removed PSE from the prohibited list. Prior to 2004 CATH was a forbidden substance and it is still one. CATH is included on the WADA prohibited list in the group of stimulants. The results of a doping control concerning PSE conducted in the Department of Anti-Doping Research of Institute of Sport in Warsaw in the years 2001-2003 and 2004-2007 have been compared. Moreover, several dozen of urine samples collected from the patients taking OTC drugs with PSE have been analysed. In these samples the concentration of PSE and CATH has been estimated. The results of this study have shown that athletes were using PSE frequently and in high doses between 2004 and 2007 when this substance was permitted by WADA. It is possible that athletes can obtain a positive result of doping control with CATH after the use of PSE.

The effect of endotoxin administration on cytokine production in obstructive jaundiced rats.

The aim of this study was to evaluate the effect of lipopolysaccharide (LPS) administration, which mimics a surgical intervention, on the immune status of obstructive jaundiced (OJ) and sham-operated control rats. Rats were given 20 microgram LPS intraperitoneally on day 13 following bile duct ligation or sham surgery. We determined serum levels of tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) on day 14 after surgery, and spontaneous as well as LPS-induced production of these cytokines in splenocyte and peritoneal exudate cell (PEC) cultures. We found that IL-6, but not TNF-alpha, serum concentrations were significantly elevated (4-fold) in OJ rats treated with LPS compared with LPS-untreated OJ rats. In sham-operated rats the differences between the respective groups were not significant. The production of TNF-alpha by splenocyte and PEC cultures was depressed in OJ rats treated with LPS; in particular, a very deep decline was observed in the case of spontaneous TNF-alpha production in PEC cultures. In contrast, TNF-alpha production in LPS-untreated and LPS-treated sham-operated rats did not differ. In the case of IL-6 production by splenocytes and PEC cultures, we observed a significant suppression of this cellular function in both OJ and sham-operated rats treated with LPS when compared with the respective controls. In conclusion, the results indicate that the already depressed cytokine production in OJ rats leads to even deeper hyporeactivity following LPS challenge. Lack of TNF-alpha suppression upon LPS treatment in sham-operated rats suggests that surgery-elicited hyporeactivity is mediated by a different mechanism than that leading to immune hyporesponsiveness in OJ. Our findings may explain the relatively high mortality rates observed of OJ patients subjected to surgery.

[Genetic changes and clinical management in familial hypertrophic cardiomyopathy]. / Zmiany genetyczne a obraz kliniczny rodzinnej kardiomiopatii przerostowej.

Hypertrophic cardiomyopathy (HCM) is phenotypically and genotypically heterogeneous disease of heart. Nine chromosomal loci responsible for this condition have been identified: beta-myosin heavy chain, essential and regulatory myosin light chains, troponin T and I subunits, alpha-tropomosin, cardiac myosin binding protein C, cardiac actin and titin. These genes code for proteins involved in the contraction mechanism or in the control of contraction, therefore HCM has been classified as a disease of cardiac sarcomere. Over 107 mutations have been identified. More then half of them have been detected in the beta-myosin heavy chain gene (beta-MHC). Some mutations in beta-MHC gene are associated with a benign prognosis, other are associated with high incidence of sudden cardiac death (SCD) and severe hypertrophy. Mutations in myosin binding protein C are associated with mild, delayed expression of cardiac hypertrophy and benign prognosis. Mutations in cardiac troponinT are associated with a mild degree of hypertrophy but a high incidence of SCD. Study of genes responsible for HCM will assume role in the context of clinical management of HCM, in particular regarding diagnosis and prognosis patients and families with HCM.

The impairment of wound healing process is correlated with abnormalities of TNF-alpha production by peritoneal exudate cells in obstructive jaundiced rats.

The wound healing process and production of tumour necrosis factor alpha (TNF-alpha) by peritoneal cells of 7-day and 14-day obstructive jaundice (OJ) and sham-operated rats were investigated. In the study the skin wound breaking strength was measured. In addition such histological and biochemical parameters as fibroblast and endothelial cell proliferation, inflammatory cell infiltration and hydroxyproline content were evaluated in polyurethane sponge discs implanted subcutaneously into rats. TNF-alpha production by peritoneal exudate cells (PEC), both spontaneous and lipopolysaccharide (LPS)-induced was determined by a bioassay. In OJ rats the process of both early as well as late phase of healing was impaired. The breaking strength of skin wound was decreased, the fibroblast and endothelial cell proliferation and collagen deposition, as well as hydroxyproline content were diminished. In 7 day OJ the numbers of inflammatory cells in the implants were lowered with a subsequent slight increase on day 14 of OJ. The spontaneous and LPS induced TNF-alpha production by PEC were significantly higher in 7 day OJ as compared with sham-operated controls. On day 14 of OJ the LPS-induced TNF-alpha level was, in contrast, much lower and did not differ much from the spontaneous TNF-alpha production. We conclude that the impairment of wound healing in OJ results from disturbances in functioning of the immune system caused by systemic endotoxaemia.

[Adhesive glycoproteins of the extracellular matrix]. / Adhezyjne glikoproteiny macierzy zewnatrzkomórkowej.

Glycoproteins of the extracellular matrix and basement membrane play crucial role in cell recognition, adhesion, migration and proliferation. Morphogenesis and tissue and organ development in embryo, as well as healing processes in adults are dependent on the interactions of extracellular proteins with cell surface receptors. In this paper the structure and function of major adhesive proteins: fibronectin, tenascin, laminin and nidogen are reviewed. Recent data on the role of small leucine-rich and modular proteoglycans, such as decorin, biglycan, fibromodulin, versican, perlecan, neurocan, agrecan, agrin and other are given. Modifications of the expression and localization of extracellular glycoproteins in pathology are also presented.

Platinum complexes and pyruvate kinase activity.

The interaction of platinum complexes with bovine heart pyruvate kinase (PK) was studied by absorption, CD, fluorescence spectroscopy and enzymic activity test. Our results showed that activity of PK was reduced by cis-DDP and potassium tetrachloroplatinate in a time-and concentration dependent manner. Cis-DDP was less effective than K2PtCl4 in reducing PK activity. The native enzyme showed well defined negative Cotton effect at 222 and 208 nm indicating the presence of alpha-helical and beta structure. Platinum binding lowered the Cotton effect in this region by about 10-20% and 30-50% for the system with cis-DDP and K2PtCl4, respectively. Fluorescence study showed that platinum binding quenched tryptophan fluorescence suggesting that binding occurs at the tryptophan residue or its proximity. PK modifications induced by platinum binding would result in a greater resistance to denaturing agents.

[Metabolism of nicotine--mechanism and clinical effects of toxicity]. / Metabolizm nikotyny--mechanizm i kliniczne efekty toksycznego dzialania.

Nicotine is a major constituent of tobacco and exerts a number of physiological effects. Metabolism of nicotine in animal and human tissues is reviewed. Clinical considerations regarding the problems of toxic effects of nicotine in human and molecular basis of carcinogen mechanism of nicotine derivatives are discussed.

Structure and expression of the cDNA for the C isozyme of phosphofructo-1-kinase from rabbit brain.

Rabbit brain contains three phosphofructo-1-kinase (PFK) isozymic subunits designated A, B, and C. The primary structures of the first of these two isozyme types have been determined previously. The isozyme C of rabbit brain was isolated by immunoaffinity chromatography and subjected to proteolytic and chemical digestion. A large number of peptides were sequenced, the total number of amino acids identified being equal to about 80% of the total structure. The sequence of the cDNA derived from brain mRNA for C isozyme was determined from polymerase chain reaction fragments synthesized using oligonucleotides designed on the basis of the peptide sequences. The deduced size of the C isozyme was 86,371 Da, slightly larger than PFKs described previously. The amino acid sequence identity with the rabbit A isozyme was 68.9% and a range of identity to other sequenced mammalian PFKs was 67-69%. Using these data plus previously published data on chemical modification, assignments of the 6 organic ligand binding sites of PFK were inferred. The full-length cDNA was cloned into and expressed in Escherichia coli. Phosphofructokinase C was purified to homogeneity from the bacterial extracts.

[Proteins modified in the nonenzymatically glycosylation reaction (AGE-proteins)--new markers for diabetes?]. / Bialka zmodyfikowane w reakcji nieenzymatycznej glikozylacji (AGE-proteiny)--nowe markery w cukrzycy?

Paper present a recent review on the formation and clinical significance of advanced glycosylation end products, produced in nonenzymatically glycosylation, called Maillard reaction. The special attention was paid to AGEs role in diabetic and aging processes. Instant of occurring of AGEs in circulation or increase of AGE receptor concentration are many years faster than clinical pathology of vessels, nervous or kidneys connect with diabetes or aging. May be in the future it will be possible to decrease the consequence of Maillard reaction by using pharmacology drugs.

[Surgical treatment of ventricular tachycardia in patients with post-infarction aneurysms]. / Leczenie operacyjne czestoskurczów komorowych u chorych z tetniakami pozawalowymi serca.

4 patients (P) with recurrent, sustained ventricular tachycardia (VT) resistant to medical treatment, underwent surgery for cure of this arrhythmia. Each P had episodes of VT lasting 30 or more seconds, 3 of them had episodes of ventricular fibrillation. In all cases rhythm disturbances were secondary to post myocardial infarction aneurysm. Coronary angiography showed in all P total occlusion of LAD, in 2 cases significant lesion in RCA were found. 1 P had lung cancer. All P underwent aneurysmectomy and an excision of the altered endocardium by Harken's method. The endocardial excision was performed without endocardial mapping. 2 P had concomitant CABG to RCA. In the P with lung cancer lobectomy was performed. There were 2 ++non-arrhythmic death. The P with lung cancer died because of sepsis due to lung abscess. One P died because of heart failure (preoperative EF 10%), 6 months after the surgery. The 2 survivors remained free of VT during a follow-up period 8 months. In conclusion, endocardial excision by Harken's method is efficient in treating recurrent sustained VT, resistant to medical treatment, in patients with post myocardial infarction aneurysm. The surgical procedure can be performed without intraoperative endocardial mapping.

[Analysis of causes for hospitalization in the pediatric department of a regional hospital in Trzcianka in the years 1981-1985]. / Analiza przyczyn hospitalizacji w oddziale dzieciecym szpitala rejonowego zoz w trzciance w latach 1981-1985.

The authors analysed 2,575 case histories of children hospitalised in 1981-1985 at pediatric department of regional hospital in Trzcianka (Pila region) taking to account: age, sex, permanent residence, duration of stay at a hospital, disease causing hospitalisation. Within the period studied both decrease of number of neonates (1-6 months old) and young children of (1-3 years old) admitted to the hospital and a tendency toward shortening time of hospitalisation were found. Among causes of hospitalisation the first were respiratory tract infections (88.3% of all admissions in 1981 but 69.8% in 1985), and acute diarrheas (22.1% of all admissions in 1981, 17.1% in 1985).

The sites of phosphorylation of rabbit brain phosphofructo-1-kinase by cyclic AMP-dependent protein kinase.

The three isozymic subunits of phosphofructo-1-kinase present in rabbit brain and designated A, B and C were phosphorylated in vitro by cyclic AMP-dependent protein kinase with 32P-labeled ATP. Limited digestion of the labeled enzymes with trypsin or with Staphylococcus aureus V8 proteinase led to the solubilization of radiolabeled peptides derived from the three isozymic subunits. Limited digestion by V8 proteinase was accompanied by a slight reduction in the apparent sizes of the subunits, indicating that the phosphorylated sites are located near either the amino or carboxyl termini of the protein. V8 proteinase digestion led to no change in the maximal activity of the enzyme but did abolish sensitivity to ATP inhibition. The phosphopeptides of the tryptic and the V8 digests were purified by chromatography and their amino acid sequences were determined and compared to the previously established sequence from rabbit muscle isozyme A. PFK-A E H I S R K R S G E A T V PFK-B H V T R R S L S M A K G F PFK-C V S A S P R G S Y R K F L In each instance, the phosphorylated serine, underlined in the above sequences, was found to be one or two residues toward the C-terminus of one or more basic residues. No other similarities in structure were noted.