[Biomarkers after resuscitation : Relevance in daily clinical practice for prognosis estimation and definition of therapeutic goals]. / Biomarker nach Reanimation : Relevanz im klinischen Alltag für Prognoseeinschätzung und Therapiezieldefinition.

BACKGROUND: The assessment of the neurological prognosis after cardiac arrest should be made using a multimodal approach involving clinical, physical and laboratory findings. Here, biomarkers are of high importance. The reliable prognostication has far-reaching consequences for the patient on the further course of therapy and rehabilitation. OBJECTIVES: Which biomarkers help in prognosis estimation and therapy target definition and are currently used in daily clinical practice? MATERIALS AND METHODS: Presentation of the multimodal approach for prognosis generation in patients after resuscitation with hypoxic-ischemic encephalopathy with special consideration and discussion of various biomarkers. RESULTS AND CONCLUSION: Neuron-specific enolase (NSE) is the best-established predictive biomarker in patients with hypoxic-ischemic encephalopathy after cardiac arrest. In combination with other methods (clinical examination, physical testing) and considering possible interfering factors (hemolysis, tumor diseases), NSE is used after 48-72 h with a cutoff value of 90 ng/ml. Most other biomarkers have so far only been studied in smaller groups or individual studies and thus cannot currently be routinely used outside of studies.

Loss of heterozygosity on chromosome 11q22-23 in melanoma is associated with retention of the insertion polymorphism in the matrix metalloproteinase-1 promoter.

Matrix metalloproteinase-1 (MMP-1, collagenase-1), which degrades interstitial collagen, is expressed at high levels by some tumor cells and is thought to enhance their invasiveness and metastatic potential. We recently described a common single nucleotide insertion polymorphism (2G allele) at -1,607 bp in the promoter of the MMP-1 gene that creates a binding site for the ETS family of transcription factors, and that is associated with enhanced transcription of this gene and increased enzyme activity. Allelic loss at the MMP-1 locus on chromosome 11 occurs in many tumors including melanoma, an invasive and aggressive cancer. We hypothesized that although loss of either the 1G or 2G allele from 1G/2G heterozygotes is random, retention of the transcriptionally more active 2G allele would favor tumor invasion and metastasis. As a result, a higher proportion of metastases would contain the 2G genotype than the 1G genotype. We report here the development of quantitative methods for assessing allelic loss at the MMP-1 locus, and demonstrate that 83% of the metastatic melanomas with loss of heterozygosity at this locus retained the 2G allele. This supports the hypothesis that retention of the 2G allele favors tumor invasion and metastasis in melanoma.

Improved blood cellular biocompatibility with heparin coated circuits during cardiopulmonary bypass.

BACKGROUND: The clinical use of heparinized surfaces in the extracorporeal circuit was studied to find out if there was any blood cell rheologic benefit to support its use in routine low risk cardiac surgery. METHODS: In a prospective single blind study, 39 patients were operated upon with the heart lung machine for angina pectoris by coronary bypass grafting and were randomized to a control group or a heparin group. Blood cell rheology was analysed using the St. George filtrometer where damage to the red blood cells and white blood cells was estimated by assessing deformability reductions, transit, time increases and clogging rate and clogging particle changes. RESULTS: At the end of cardiopulmonary bypass, in the heparin group, the red cell filterability (rFR) and the white cell filterability (WFR) were 8% better than in the control group (p=0.0079 and p=0.027 respectively). The red cell transit time was 19% slower in the control group (p=0.0351). The red cell clogging rate (RCR) and clogging particles (RCP) were significantly lower in the heparin group (p=0.0212 and p=0.0409 respectively. The white cell clogging rate (WCR) and clogging particles (WCP) showed a similar pattern. CONCLUSIONS: In spite of these significant differences the clinical outcome was similar in the groups. Thus heparin coating of the extracorporeal circuit reduces blood cell rheologic damage significantly in low risk patients undergoing routine bypass surgery for angina but this use did not lead to any clinical benefit postoperatively. Therefore the use of such circuits for routine low risk cardiac surgery cannot be recommended.

Heparin-coated circuits reduce occult myocardial damage during CPB: a randomized, single blind clinical trial.

OBJECTIVES: Cardiopulmonary bypass is associated with a diffuse systemic inflammatory response that can cause considerable morbidity, including organ dysfunction and bleeding. Heparin-coated circuits have been shown to give a reduced inflammatory response with clinical benefits during open-heart surgery. However, the effects on lipid peroxidation, neutrophil activation and myocardial ischemic damage in the human have remained unknown. METHODS: In a randomized single blind trial, complement activation, neutrophil counts, malondialdehyde, and cardiac enzymes were studied in 39 patients undergoing open-heart surgery. Two groups were perfused with cardiopulmonary bypass circuits with (n=20) or without heparin-coating (n=19). RESULTS: The different complement factors (C3, C4, C3d, C3d/C3 and the C-function), neutrophil levels, MDA and the cardiac enzyme levels were comparable before CPB was started and significantly increased in both groups during bypass. There were significant intergroup differences in the neutrophil levels and MDA after reperfusion (P<0.0001). Furthermore, significant positive correlations between the lipid peroxidation, expressed as MDA levels, and the levels of neutrofils and the cardiac enzyme, CK-MB were seen within the groups. CONCLUSIONS: Heparin coated circuits did lead to a decreased neutrophil response and MDA level. The correlations between CK-MB and neutrophil and MDA levels suggest neutrophil activation leading to lipid peroxidation that may influence myocardial damage. Heparin coating improved biocompatibility and was associated with less occult myocardial ischemic damage in patients undergoing open heart surgery.

Familial breast cancer risk assessment.

A family, with a strong history of dominant breast and ovarian cancer, is described. Using highly polymorphic microsatellite markers within the BRCA1 breast cancer gene on chromosome 17q21; three affected sisters, their father and a paternal second cousin once removed, are shown to share the same "abnormal" haplotype. Because of this informative linkage, the carrier status of the unaffected siblings can be established by determining whether they inherited their father's "normal" or "abnormal" haplotype. Presymptomatic diagnosis is important in decisions regarding prophylactic surgery or follow-up care. However, the widespread general population presymptomatic DNA testing of breast cancer is currently not recommended because of inherent problems in the sensitivity and specificity of DNA testing.

Minibands and chromosome structure. A theory.

Bright and dark field microscopy were used to examine interphase nuclei and RBG-banded human chromosomes. In dark field microscopy both chromosomes and interphase chromatin appeared to be composed of similar independent structures alternating between bright and dull light intensity. The position, colour, and number of these structures may support the theory that the transition from interphase chromatin to metaphase chromosomes, takes place without any dramatic changes in local chromatin structure. Although the resolution power of bright and dark field microscopy prevent a direct proof for the miniband model, our observations in combination with other data may add further support to this particular model of chromosome organization.

Cellular pharmacology of phosphorothioate homooligodeoxynucleotides in human cells.

The uptake and distribution of phosphorothioate oligodeoxynucleotides by human cells were studied using 35S-labeled 28-mer phosphorothioate oligodeoxycytidine [S-(dC)28]. Accumulation of intracellular S-(dC)28 was found to be higher in the carcinoma cells (grown in monolayers) than in the leukemia cells (grown in suspension culture). A hepatoma cell line transfected with hepatitis B virus, 2215, was chosen for further studies. The uptake of S-(dC)28 was partially dependent on temperature and energy. The intracellular concentration was significantly higher than that in the medium and the amount accumulated was dependent on the extracellular concentration. It appears that the uptake of S-(dC)28 involves mechanisms of both fluid-phase pinocytosis and adsorptive endocytosis. Neither oligonucleotides nor 5'-phosphorylated nucleotides inhibited S-(dC)28 uptake. Unlike horseradish peroxidase, which was primarily associated with endosomes once it was taken into the cell, S-(dC)28 was found to be present in both nuclear and cytoplasmic fractions. Efflux of S-(dC)28 from the cell was multiphasic; a trapping mechanism that could be due to a potent interaction of S-(dC)28 with cellular proteins was implicated. This trapping mechanism could be responsible for the lack of biological activity such as cytotoxicity and antisense activity of phosphorothioate oligodeoxynucleotides in some human cells.

Anabaseine: venom alkaloid of aphaenogaster ants.

Anabaseine, a tobacco alkaloid, is identified as a poison gland product in Aphaenogaster ants, in which it functions as an attractant.