Journal des biomarqueurs moléculaires et du diagnostic

The buccal micronucleus cytome assay in exfoliated buccal cells is utilized as biomarkers for DNA damage, cell death and basal cell frequency. It offers great opportunity to evaluate genotoxicity by the way of quantifying mean frequencies of micronuclei, binucleated cell, broken egg, karyolysis, karyorrhexis, pycknosis and condensed chromatin. This assay is sensitive, minimally invasive, simple, cheap, easy and fast. It has precision and statistical power obtained from scoring large number of cells. Micronucleus assay has been extensively used to assess genetic damage due to lifestyle characteristics, occupational exposure, diseases and environmental risk. It also has applications in human biomonitoring, ecotoxicology, cancer risk assessment, pharmaceutical drug testing and the impacts of dietary micronutrients and micronutrient combinations on DNA damage. Fluorescence in situ hybridization is a valuable addition to micronucleus assay as combination of both enables us to characterize the genetic contents of the micronuclei. The present article reviews and updates on usefulness of buccal micronucleus cytome assay as a biomarker. It gives a detailed description of the methodology of buccal micronucleus test and analysis of the results. We also discussed the criteria for identification and classification of nuclear anomalies. We have also proposed the future directions namely high-throughput automation for further enhancing the reliability of micronucleus assay to be applicable on large scale experimental and epidemiological studies. It would help in overcoming many of the problems caused by inter-observer variability in evaluation of slides.

Objectives: Interleukin-33 (IL-33) is a new member of the IL-1 cytokine family, which is thought to be involved in the pathogenesis of various inflammatory diseases, through its soluble receptor ST2. There is increasing evidence that inflammation is a relevant player in structural atrial remodeling that represents the main mechanism for atrial fibrillation (AF) persistence. This study was designed to investigate the state of IL-33/ST2 axis serum concentrations in patients with persistent AF.

Design and Methods: We investigated the concentrations of IL-33, its soluble ST2 receptors, and high-sensitivity C-reactive protein levels (hsCRP) in the sera of 92 patients with persistent atrial fibrillation, and 68 controls.

Results: Serum concentrations of IL-33, sST2, and hsCRP were all significantly elevated in patients with persistent AF compared to controls (P <0.0001 for all). Moreover, serum IL-33 concentrations was positively correlated with the inflammatory marker hsCRP (r=0.606, P =0.002).

Conclusion: These preliminary results may support the role of inflammation in AF pathogenesis and IL-33/sST2 axis may be involved in the inflammatory process in AF.

Background: Measurement of faecal calprotectin (FCP) concentrations is a theoretically justified laboratory test used in lieu of quantitative evaluation of neutrophils penetrating into the intestinal lumen. This parameter is organspecific and identifies intestinal inflammation.

Aim: Search for individual interpretation of the cut-off values for the FCP test, taking into consideration different age groups of patients and causes of intestinal inflammation.

Methods: Review of the literature data about the calprotectin concentrations in the meconium and faeces of neonates, infants, older children and adults.

Results: FCP levels in meconium and faeces of infants are significantly higher than the concentration in older children and adults. In adults FCP is a simple and sensitive screening test for inflammatory bowel disease, also used in monitoring of disease activity and treatment efficacy. However, the diagnostic significance of high FCP levels in healthy infants and young children remains unclear.

Conclusions: FCP levels in the first year of life are higher than in adults and probably reflect ‘physiological’ intestinal inflammation associated with the adaptation of the neonate’s intestine to life outside the uterus. On the other hand, high CP levels in meconium may indicate the important role played by neutrophils in the fetal intestine during its development.

The exploitation of DNA polymorphisms by an ever-increasing number of molecular marker technologies has begun to have an impact on animal and plant genome research and breeding. A large number of different molecular techniques for genetic diversity analysis are available and each of them differs in its informational content. These techniques not only helped in understanding the different structure and behavior of species genome, but also revolutionized and modernized the ability to characterize genetic variation. Identification of marker linked to useful traits has been based on complete linkage maps and bulked segregation analysis. Markers are therefore used to identify traits, their inheritance and characterize germplasm.

 Introduction: Iron over load estimation was attempted by MRI R2* and serum ferritin, in non -transfusion dependent HbE Beta Thalassaemia patients.

Methods: Seventy three patients selected on definite criteria were scanned and tested. Average age of the patients was 20.8 ± 11.14 years, 33 females and 40 male. Average hepatic iron concentration was determined to be 11.09 ± 11.74 mg/g of dry liver tissue and average serum ferritin was 972.44 ± 1121.51 ng/ml.

Results: 81% of the patients had hepatic iron concentration more than 2 mg/g of dry liver tissue, whereas only 41% had serum ferritin level >500 ng/ml, irrespective of age. The hepatic iron overload pattern was heterogenous, where age and sex were not well determined variable predictors of such iron overload. Though, higher hepatic iron load was seen more predictably in patients above the age of 10 years and LIC of 5 mg/g of dry tissue correlated with serum ferritin level of approximately 400 ng/ml.

Conclusion: It can be concluded that non transfusion dependence does not ensure non iron overload status and such patients should be scanned by using MRI R2* sequence to determine their hepatic iron status and start on excess iron chelation therapy if felt required.

Objective: To investigate the pharmacokinetics of cycloserine in healthy rat blood and lung tissues.

Method: Healthy rat blood and lung tissues were sampled in vivo by microdialysis sampling technique simultaneously. The concentrations of cycloserine in both blood and lung tissues were measured by high performance liquid chromatography mass spectrometry. All date were analyzed by WinNonlin software

Results: The maximum concentration of free cycloserine in blood and lung tissue was (10.61 ± 2.42) mg/L and (1.53 ± 1.71) mg/L at 1 h and then both continued to decline. After administration the concentrations of free cycloserine in the blood has been higher than the concentrations in lung tissues. The area under the concentration curve (AUC) of free cycloserine was (33.53 ± 6.51) h.mg-1.L-1 in blood and (4.49 ± 2.08) h.mg-1.L-1 in lung tissues.

Conclusion: Microdialysis sampling technique combined with high performance liquid chromatography mass spectrometry / mass spectrometry can be accurately and objectively reflect the drug in the blood and tissues of the pharmacokinetic characteristics of cycloserine. Concentrations in lung tissues were significantly lower than concentrations in blood.