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Clinical Biochemistry (v.39, #3)

Editorial Board (pp. co2).

Genome-wide detection of allelic imbalance in renal cell carcinoma using high-density single-nucleotide polymorphism microarrays by Ching-Wan Lam; Ka-Fai To; Sui-Fan Tong (pp. 187-190).
Renal cell carcinoma (RCC) appears in both a sporadic form and a hereditary form. Eighty-five percent of sporadic RCCs are of the clear-cell histologic type. The cytogenetic analysis of RCCs has revealed several recurring sites of chromosomal aberrations (non-disjunction, deletion or mitotic recombination) including segments of loss of heterozygosity (LOH) identifiable by polymorphic markers. In this pilot study, we performed a comprehensive genome-wide scan to identify LOH sites of RCCs in three Chinese patients using high-density single-nucleotide polymorphism microarrays (HuSNP arrays).Three sporadic clear-cell RCCs specimens were diagnosed histologically. Tumor genomic DNA was extracted from paraffin-embedded sections after microdissection to avoid gross contamination by non-tumor cells. Germline DNA was obtained from paired normal adjacent tissues. Affymetrix HuSNP mapping assay was performed according to the manufacturer's instructions.Using high-density single-nucleotide polymorphism microarrays, we were able to identify the previously described and new LOH sites in RCCs of the three Chinese patients.The high-density single-nucleotide polymorphism microarrays and assays offer significant operating cost benefits in sample preparation, processing, and data analysis for identification of LOH sites in cancer samples. In contrast to the typical microsatellite genotyping strategy, the entire genome scan is completed in one experiment taking less than 2 days.

Keywords: High-density single-nucleotide polymorphism microarrays; Renal cell carcinoma; Allelic imbalance; Loss of heterozygosity

Paraoxonase 1 gene Q192R polymorphism affects stroke and myocardial infarction risk by Larry Baum; Ho Keung Ng; Kam Sang Woo; Brian Tomlinson; Timothy Hudson Rainer; Xiangyan Chen; Wing Sze Cheung; Daniel Kam Yin Chan; G. Neil Thomas; Cindy See Wai Tong; Ka Sing Wong (pp. 191-195).
Paraoxonase (PON1), an enzyme associated with high-density lipoprotein (HDL) particles, inhibits oxidation and atherogenesis. We sought to investigate the association of the PON1 Q192R polymorphism with stroke and heart disease.In a case control study, we genotyped 242 ischemic stroke, 231 myocardial infarction (MI), and 310 healthy control subjects, all Chinese. R-containing genotypes ( R+) were associated with vascular disease, OR=1.5, P=0.03. RR was increased in MI patients who were either smokers (OR=3.1, P=0.01), male, or younger than 60. R+ but not RR genotypes were increased in stroke patients, particularly large artery type (OR=2.6 and P=0.02 for R+, OR=1.0 for RR) or among smokers. The relative dearth of RR in stroke might be due to earlier MI or death in at-risk people, such as smokers. R+ genotypes were increased with stroke in hypertensive (OR=2.1, P=0.02) but not normotensive (OR=1.0) subjects. PON1 192R+ genotypes were associated with stroke and MI, particularly in subsets of patients, in patterns suggesting a possible survivor effect.

Keywords: Paraoxonase; Polymorphism; Ischemic stroke; Myocardial infarction

High rate of detection of subtelomeric aberration by using combined MLPA and subtelomeric FISH approach in patients with moderate to severe mental retardation by Albert C.F. Lam; Stephen T.S. Lam; Kent K.S. Lai; Tony M.F. Tong; T.C. Chau (pp. 196-202).
(1) To evaluate the prevalence of subtelomeric deletion in moderate to severe mental retardation population, (2) to assess the feasibility and cost-effectiveness of combined methodology in routine workup of this sub-population.Twenty unrelated patients using strict selection criteria were recruited for the study from the Clinical Genetic Service. Patients were initially screened by Multiplex Ligation-dependent Probe Amplification (MLPA) for subtelomeric imbalance followed by FISH analysis for anatomical integrity. This is then followed by parental subtelomeric FISH analysis.Three subtelomeric deletions were identified. They were Deletion 1p36, Deletion 1q44 and Deletion 10q26; these were previously unidentified by conventional technique.The prevalence of subtelomeric deletion in our cohort of moderate to severe mental retardation patients is consistent with published findings of around 10%. The figure is on the higher side if more stringent criteria is used. The combination of strict clinical criteria, MLPA and selective subtelomeric FISH was shown to be feasible and cost-effective.

Keywords: Moderate to severe mental retardation; Subtelomeric deletions; MLPA; Subtelomeric FISH; MR, mental retardation; FISH, fluorescence in situ hybridization

Metabolomic and bioinformatic analyses in asphyxiated neonates by Ching Yan Chu; Xin Xiao; Xiao Guang Zhou; Tze Kin Lau; Michael Scott Rogers; Tai Fai Fok; Lap Kay Law; Chi Pui Pang; Chi Chiu Wang (pp. 203-209).
We tested the application of bioinformatic algorithms in studying the metabolomic profiles of neonatal urine samples with clinical evidence of severe asphyxia at birth and subsequent neurodevelopmental handicap.The clinical outcomes of 256 newborns that required direct admission to neonatal intensive care unit for respiratory support or did not require direct admission were studied. Urinary metabolite profiles were measured by high throughput mass spectrometry and analyzed by bioinformatic methods.We found a positive relationship between suppressed biochemical networks involved in macromolecular synthesis and birth asphyxia associated with significant neonatal oxidative stress and morbidity. The metabolomic discriminators between good neonatal outcome and poor neonatal outcome were established using hierarchical clustering analysis. Concentrations of eight urinary organic acids in distinct biochemical pathways were elevated and significantly associated with the prognosis of neurodevelopmental handicap with high sensitivity and specificity: ethylmalonate, 3-hydroxy-3-methylglutarate, 2-hydroxy-glutarate and 2-oxo-glutarate were associated with good neonatal outcome, whereas glutarate, methylmalonate, 3-hydroxy-butyrate and orotate were associated with poor outcome.The data demonstrated the potential application of bioinformatics methods in this metabolomic study and proved its clinical relevance.

Keywords: Asphyxia neonatorum; Metabolomics; Organic acids; Bioinformatics

Analysis of GABRB2 association with schizophrenia in German population with DNA sequencing and one-label extension method for SNP genotyping by Zhiliang Yu; Jianhuan Chen; Haifeng Shi; Gerald Stoeber; Shui-Ying Tsang; Hong Xue (pp. 210-218).
Schizophrenia (SCZ) is a complex mental disease that affects approximately 1% of the population. In this study, six SNPs in GABRB2 were genotyped for a case–control association study with the cycloid psychosis subtype of SCZ in the German population using two methods for SNP genotyping.The SNPs were genotyped by direct DNA sequencing, as well as a novel one-label extension method. The results were analyzed for association with SCZ.Significant association was found for SNPs rs1816071 and rs1816072 with SCZ susceptibility. This is consistent with our previous finding of association of SNPs in GABRB2 with SCZ susceptibility in Han Chinese. There was a total agreement between the genotyping results from one-label extensions and the results of direct DNA sequencing, thus validating the accuracy of the one-label extension method of SNP genotyping.

Keywords: Schizophrenia; GABA(A) receptor; Association study; Genotyping

Prenatal detection of a de novo Yqh-acrocentric translocation by Lucy K.L. Ng; Yvonne K. Kwok; Linda Y.F. Tang; Paulina P.Y. Ng; A. Ghosh; Elizabeth T. Lau; Mary H.Y. Tang (pp. 219-223).
To identify the extra chromosomal material on 46,XX,21p+ for prenatal diagnosis.Conventional cytogenetic studies using GTG (G bands by trypsin using Giemsa) and CBG (C bands by barium hydroxide using Giemsa) techniques were performed on chromosomes at metaphase obtained from cultured amniocytes and parental blood lymphocytes. Molecular cytogenetic techniques, QF-PCR (quantitative fluorescent polymerase chain reaction), FISH (fluorescent in-situ hybridization), and DA-DAPI (Distamycin A and 4,6-diamino-2-phenylindole) staining, were then used to clarify the extra material present on fetal chromosome 21 p.The extra material on fetal chromosome 21 p has originated from Yqh, most likely at PAR2 (the secondary pseudoautosomal region). The karyotype should be 46,XX,der(21)t(Y;21)(q12;p13)de novo.ish der(21)t(Y;21)(q12;p13) (EST Cdy16c07+).This case demonstrates the usefulness of molecular techniques in the investigation of rare chromosomal rearrangements.

Keywords: Yqh-acrocentric translocation; PCR; QF-PCR; FISH; DA-DAPI; PAR2

Molecular characterization of the developmental gene in eyes: Through data-mining on integrated transcriptome databases by K.W. Choy; C.C. Wang; A. Ogura; T.K. Lau; M.S. Rogers; K. Ikeo; T. Gojobori; L.Y. Tang; D.S.C. Lam; T.K.H. Chung; C.P. Pang (pp. 224-230).
Our aim was to utilize publicly available and proprietary sources to discover candidate genes important for ocular development.The collated information on our 5092 non-redundant clusters was grouped and functional annotation was conducted using gene ontology (FatiGO) for categorizing them with respect to molecular function. The web-based viewer technological platform (H-InvDB) was employed for transcription analyses of in-house high quality fetal eye Expressed Sequence Tags (ESTs). Eye-specific ESTs were also analyzed across species by using EMBEST.According to adult eye cDNA libraries, nucleic acid binding and cell structure/cytoskeletal protein genes were the most abundant among the ESTs of fetal eyes. Using cDNA assembly in H-InvDB, 20 (80%) of the 25 most commonly expressed genes in the human eye are also expressed in extraocular tissues. The crystalline gamma S gene is highly expressed in the eye, but not in other tissues. We used EMBEST to compare human fetal eye and octopus eye ESTs and the expression similarity was low (1.6%). This indicated that our fetal eye library contains genes necessary for the developmental process and biological function of the eye, which may not be expressed in the fully developed octopus eyes. The human fetal eye cDNA library also contained highly abundant eye tissue genes, including αA-crystallin, eukaryotic translation elongation factor 1 alpha 1 ( EEF1A1), bestrophin ( VMD2), cystatin C, and transforming growth factor, beta-induced ( BIGH3).Our annotated EST set provides a valuable resource for gene discovery and functional genomic analysis. This display will help to appreciate the strengths and weaknesses of the different technological platforms, so that in future studies the maximum amount of beneficial information can be derived from the appropriate use of each method.

Keywords: Eye; ESTs; Human; Octopus; Bioinformatics

Molecular diagnostics of genetic eye diseases by Bao Jian Fan; Pancy Oi Sin Tam; Kwong Wai Choy; Dan Yi Wang; Dennis Shun Chiu Lam; Chi Pui Pang (pp. 231-239).
Eye diseases can be simple or complex, and mostly of heterogeneous molecular genetics. Some eye diseases are caused by mutations in a single gene, but some diseases, such as primary open angle glaucoma, can be due to sequence variations in multiple genes. In some diseases, both genetic and epigenetic mechanisms are involved, as was recently revealed in the mechanism of retinoblastoma. Disease causative mutations and phenotypes may vary by ethnicity and geography. To date, more than a hundred candidate genes for eye diseases are known, although less than 20 have definite disease-causing mutations. The three common genetic eye diseases, primary open angle glaucoma, age-related macular degeneration, and retinitis pigmentosa, all have known gene mutations, but these account for only a portion of the patients. While the search for eye disease genes and mutations still goes on, known mutations have been utilized for diagnosis. Genetic markers for pre-symptomatic and pre-natal diagnosis are available for specific diseases such as primary open angle glaucoma and retinoblastoma. This paper reviews the molecular basis of common genetic eye diseases and the available genetic markers for clinical diagnosis. Difficulties and challenges in molecular investigation of some eye diseases are discussed. Establishment of ethnic-specific disease databases that contain both clinical and genetic information for identification of genetic markers with diagnostic, prognostic, or pharmacological value is strongly advocated.

Keywords: Genetic eye disease; Genetic testing; Molecular diagnostic service

Evaluation of an algorithm of tagging SNPs selection by linkage disequilibrium by Nelson L.S. Tang; Paul D.P. Pharoah; Suk Ling Ma; Douglas F. Easton (pp. 240-243).
Single nucleotide polymorphisms (SNPs) are the most abundant kind of genetic polymorphism in the human genome. They are important in both genetic research and genetic testing in a clinical setting, such as in the area of pharmacogenetics. In order to improve efficiency, tagging SNPs (tagSNPs) are selected in genes of interest to represent other co-related SNPs in linkage disequilibrium (LD) with the tagSNPs. Various algorithms have been proposed to identify a subset of single nucleotide polymorphisms as tagSNPs. Most algorithms of tagSNPs selection are haplotype-based, in which the spatial relationship between SNPs is considered. Currently, a more efficient cluster-based algorithm is proposed which clusters SNPs solely by a LD parameter, such as r2. Here, we evaluated the sample distribution of r2 and its effect on the cluster-based tagSNPs selection.The genotype data of 198 individual within a 500-kb region on 5q31 was used to evaluate the sample distribution of r2 and its effect on the cluster-based tagSNPs selection.It was found that the degree of variation of LD depends on the LD structure of genes.As a cluster-based tagSNPs selection algorithm does not take into account the spatial position of SNPs, a more stringent r2 threshold is required to achieve more reliable tagSNPs selection.

Keywords: Tagging; SNPs; Genetic; LD metric; Haplotype; Linkage disequilibrium

AGG interspersion analysis of the FMR1 CGG repeats in mental retardation of unspecific cause by Priscilla M.K. Poon; Qian L. Chen; Nan Zhong; Stephen T.S. Lam; Kelly Y.C. Lai; C.K. Wong; Chi-Pui Pang (pp. 244-248).
To study the AGG interspersion pattern in mentally retarded patients of unspecified cause. FMR1 CGG substructure in 104 normal and 232 mentally retarded (MR) males was determined by CGG repeat and AGG interspersion analyses. Genomic DNA of the study subjects was obtained for PCR and Southern hybridization analyses.All study subjects had less than 53 CGG repeats and none had fragile X syndrome of mental retardation. There was a significant difference ( P < 0.006) in the AGG interspersion pattern. MR males had (1) more variable internal substructures, (2) proportionally less 2 and 3 AGG but more 0 and 1 AGG, less (CGG)9AGG(CGG)9AGG(CGG)9 but more (CGG)9AGG(CGG)19 alleles and (3) a longer pure 3′ CGG repeat.Our results suggest that the MR alleles have a lesser number of interspersed AGG and a longer pure 3′ CGG repeat than the normal population. They are thus more prone to instability and expansion to long repeat lengths as in the fragile X syndrome of mental retardation.

Keywords: FMR1; AGG; CGG; Mental retardation

Gene mapping for primary open angle glaucoma by Bao Jian Fan; Dan Yi Wang; Dennis Shun Chiu Lam; Chi Pui Pang (pp. 249-258).
Primary open angle glaucoma (POAG) is a leading cause of visual impairment and blindness worldwide. To date, at least 20 genetic loci for POAG have been reported. Only 3 causative genes are identified from these loci: myocilin (MYOC), optineurin (OPTN) and WD repeat domain 36 (WDR36), which together account for less than 10% of POAG. Only a portion of POAG follows Mendelian inheritance, and a considerable fraction results from a large number of variants in several genes, each contributing small effects.Over the past 10 years, there has been vigorous research on mapping the POAG genes. The main technological approaches are functional cloning, family linkage analysis, genome-wide scan, case-control association study, and microarray analysis. Association studies found 16 genes related to POAG, but reports on glaucoma-causing effects of these genes are conflicting. Ten microarray gene expression studies related to POAG have been published. A number of genes potentially related to POAG have been identified, and they provide a good resource to select candidate genes for mutation analysis in association studies. While linkage studies remain a mainstay, the current trend is to use genome-wide association studies to map genes for POAG. This review gives an overview of the efforts in the past decade to identify the POAG genes through linkage studies, genome-wide scans, case-control association studies and microarray studies. In the near future such comprehensive studies are expected to greatly advance our understanding of the genetic basis of POAG and provide information for effective glaucoma therapy.

Keywords: Primary open angle glaucoma; Gene mapping; Linkage study; Association study; Microarray study

5′ CpG island methylation analysis identifies the MAGE-A1 and MAGE-A3 genes as potential markers of HCC by Geng Qiu; Jiancheng Fang; Yunshao He (pp. 259-266).
The change in DNA methylation patterns can be used to distinguish between normal and cancer cells. The aim of the present study was to examine the 5′ CpG island methylation patterns of the cancer-testis antigen (CT antigen) gene family, MAGE-As, in hepatocellular carcinoma (HCC), and to develop the DNA demethylation pattern as a novel tumor biomarker.We used bisulfite-sequencing PCR (BSP) to map the methylation status of the CpG site among the promoter of the MAGE-A gene family in several HCC cell lines including Hep G2, BEL7402, BEL7404, and BEL7407, and normal peripheral blood white blood cells (WBCs). According to differences of the methylation pattern between HCC cell lines and the control, methylation-special PCRs (MSP) have been developed. The developed MSPs were used to detect the paraffin-embedded slices that were pathologically diagnosed as HCC, hepatocirrhosis, hepatitis, and healthy.We found that several CpG sites among the MAGE-A1 and MAGE-A3 promoters have different methylation patterns in the HCC cell lines as compared to those in normal WBCs. Two sets of MSP primers were designed to distinguish the HCC genomic DNA and normal control cell genomic DNA as novel tumor biomarkers, and the biomarkers were validated on the archived paraffin sections of liver primary tissue. In the detection of 34 HCCs and 17 tumor-free liver tissues, the clinical sensitivity and specificity were 91.2% and 100%, respectively.Detection of aberrant methylation patterns of MAGEs CpG islands using MSP may be useful for diagnosis of HCC.

Keywords: Abbreviations; CT antigen; cancer-testis antigen; HCC; hepatocellular carcinoma; BSP; bisulfite-sequencing PCR; MSP; methylation-special PCRDNA methylation; HCC; Tumor biomarker; MSP; BSP

Contribution of VEGF and PEDF to choroidal angiogenesis: A need for balanced expressions by Jian-Ping Tong; Yu-Feng Yao (pp. 267-276).
Ocular angiogenesis may lead to visual impairment and even irreversible blindness in people of all ages worldwide. Choroidal neovascularization (CNV), a major clinical complication of ocular angiogenesis, is an important cause of vision loss that affects a large number of people. Physiological angiogenesis is tightly controlled by a balance in the expression of angiogenic and anti-angiogenic factors. While the underlying mechanism of CNV is complex, it is attributed to an upset in this balance. The vascular endothelial growth factor (VEGF) is essential in the development of CNV as one of the most potent angiogenic stimulators and vascular permeability factors. Pigment epithelium derived factor (PEDF) is a strong inhibitor of angiogenesis with high neuroprotective effects. VEGF and PEDF both possess multiple biological activities and functions that affect a large variety of tissue cells of the eye and other organs. Inappropriate expression levels are associated with many diseases involving neovascularization. This paper describes the unbalanced expressions of VEGF and PEDF as a cause of CNV. Based on the respective angiogenic and anti-angiogenic properties of VEGF and PEDF, experimental models have been devised to genetically reduce VEGF or enhance PEDF to achieve therapeutic effects. Gene therapy for CNV is promising and is under intensive research.

Keywords: VEGF; PEDF; Angiogenesis; CNV; PCV

Population distribution of high sensitivity C-reactive protein values in Aboriginal Australians: A comparison with other populations by Zhiqiang Wang; Wendy E. Hoy (pp. 277-281).
To examine the distribution of C-reactive protein (CRP) values in Aboriginal Australians and its relation to age and gender.High sensitivity CRP levels were measured in 954 Aboriginal participants aged 5–74 years. Fractional polynomial regressions were used to explore the relationship between CRP and age.CRP values changed with age and reached its lowest level around 10 years and then increased with age. Geometric means of CRP were 7.3 (95% confidence interval (CI): 6.6, 8.1) and 4.1 (95% CI: 3.7, 4.6) for female and male adults, respectively. Adjusting for age, the ratio of female to male CRP concentrations was 1.67 (95% CI: 1.45, 1.99) for adults, and 1.09 (95% CI: 0.84, 1.42) for children 5 to 19 years.CRP changes with age. Females have higher CRP values than males. CRP values in Aboriginal people are substantially higher than other populations.

Keywords: Inflammation; C-reactive protein; Aboriginal health; Cardiovascular risk factors

Thyroid function during B-vitamin supplementation of patients on antiepileptic drugs by Terje Apeland; Ole Kristensen; Roald E. Strandjord; Mohammad A. Mansoor (pp. 282-286).
Patients on antiepileptic drugs (AEDs) may have low serum concentrations of thyroxine, with or without a compensatory increment in thyroid-stimulating hormone (TSH). Furthermore, patients on AEDs often have hyperhomocysteinemia and low concentrations of vitamins B6, B2 and folate. Previously, an inverse relationship between thyroxine and homocysteine concentrations has been observed. In animals, deficiency of vitamin B6 has been found to impair the hypophyseal release of TRH. We have studied the effect of B-vitamin supplements on thyroid function in patients on AEDs.Thirty-two patients on AEDs were identified with hyperhomocysteinemia and low folate, B6 and B2. They were supplemented with pyridoxine, riboflavin and folic acid for 30 days.At baseline, the patients had low serum concentrations of free thyroxin and slightly elevated TSH. On day 30 of the B-vitamin supplements, homocysteine had decreased, however, the thyroid parameters remained unchanged.Hyperhomocysteinemic patients on AEDs have indications of hypothyroidism, however, supplementation with B-vitamins does not improve their thyroid function.

Keywords: Carbamazepine; Epilepsy; Phenytoin; Pyridoxal 5-phosphate; Riboflavin; Thyroid-stimulating hormone; Thyroxine; Triiodothyronine

Serum HDL-C levels, log (TG/HDL-C) values and serum total cholesterol/HDL-C ratios significantly correlate with radiological extent of disease in patients with community-acquired pneumonia by Omer Deniz; Ergun Tozkoparan; Halil Yaman; Erdinc Cakir; Seyfettin Gumus; Omer Ozcan; Ugur Bozlar; Cumhur Bilgi; Hayati Bilgic; Kudret Ekiz (pp. 287-292).
In several studies, it was shown that there was a marked decrease in serum levels of HDL-C during infection and inflammation in general. In particular, a decrease in the level of serum HDL-C was also shown in pneumonia. Correlations between inflammatory markers such as acute phase proteins, cytokines and serum HDL-C levels were shown. However, there are no studies indicating a correlation between serum HDL-C levels and the radiological extent of the disease (RED) in community-acquired pneumonia (CAP).We hypothesized that there could be a relationship between serum HDL-C levels and RED in CAP.A case-controlled study, including 97 patients with CAP and 45 healthy subjects, was performed. Chest X-rays of CAP patients were scored for RED, and correlations were investigated between RED scores, serum lipid parameters, the erythrocyte sedimentation rate (ESR) and serum albumin levels.The mean serum HDL-C level was lower in CAP patients than in controls. A significant and negative correlation between RED scores (REDS) and serum HDL-C levels was detected ( r = −0.64, P = 0.0001). There were also significant correlations between REDS and other lipid parameters. Significant correlations between ESR and serum HDL-C levels and between ESR and other serum lipid parameters were also found.It appears that serum HDL-C levels are generally lower in CAP cases than in healthy controls. Serum HDL-C levels and serum albumin levels might decrease and serum total cholesterol/HDL-C ratios and log (TG/HDL-C) values might increase proportionally with RED in CAP patients. These results might have some significance for individuals having long-standing and/or recurrent pneumonia and other cardiovascular risk factors.

Keywords: Pneumonia; HDL-C; Radiological extent; Log (TG/HDL-C)

Cytotoxic effects of volatile anesthetics with free radicals undergoing laparoscopic surgery by Remziye Sivaci; Ahmet Kahraman; Mustafa Serteser; Dursun Ali Sahin; Osman Nuri Dilek (pp. 293-298).
Free radicals induced by several diseases can trigger oxidative stress, leading to the production of malondialdehyde (MDA) and protein carbonyl content (CB). Volatile agents are able to increase the extent of oxidative status. However, the effects of these agents together with pneumoperitonium (Pp) have not been reported. We aimed to investigate the role of volatile anesthetics and ischemic injury during Pp on free radicals and scavenging enzymes in laparoscopic abdominal surgery.Forty patients were examined. Patients were randomly divided into four groups in order to receive sevoflurane-fentanyl (SF = 10), sevoflurane-N2O (SN = 10), desflurane-fentanyl (DF = 10), and desflurane–N2O (DN = 10), respectively. Tidal volume and ventilation frequency were kept unchanged during the operation. Intraabdominal pressure was remained constant at 12 mm Hg. Baseline values in venous blood samples were preoperatively taken and blood was also taken postoperatively at the 6th and the 24th hours. After collection of blood samples into citrate (3.5 mg/mL blood) containing glass tubes, erythrocyte sediments were prepared for the analyses. Then malondialdehyde levels, protein carbonyl content, and sulfhydryl (SH) groups were measured.The levels of MDA and protein carbonyl content were significantly higher at the 6th hour rather than the 24th hour postoperatively with desflurane anesthesia. In addition, SH groups were significantly different between the 6th hour and the 24th hour measurements ( P < 0.05). In our study, desflurane caused a statistically significant increase in MDA levels and protein carbonyl content and a decrease in SH groups. When the two groups were compared, in the case of MDA and CB values, a significant increase was observed in the 6th and the 24th hour, where there was a decrease in SH groups in the desflurane group ( P < 0.05). These parameters did not change in the sevoflurane group ( P > 0.05).We concluded that desflurane was affected by desflurane with low flow anesthesia in patients undergoing laparoscopic abdominal surgery. Significant influence on oxidative stress and antioxidant mechanics was not seen with sevoflurane anesthesia. Our studies support that oxidant and antioxidant defense mechanisms were altered in the desflurane group and this alteration improved after a combination of desflurane-N2O.

Keywords: Oxidative stress; Reactive oxygen species; Volatile anesthetics; Antioxidant enzymes

Functional polymorphism in the manganese superoxide dismutase (MnSOD) gene in patients with asthma by Lydie Izakovicova Holla; Katerina Kankova; Anna Vasku (pp. 299-302).
To investigate the possible association between functional variant Ala-9Val in the MnSOD gene and asthma in the case-control study comprising 626 Caucasian subjects.MnSOD genotypes were determined by PCR with subsequent restriction analysis by the BsaWI enzyme.Significant differences in allele frequencies between groups were not ascertained.Pursuant to these results, Ala-9Val polymorphism does not seem to be a significant predisposing factor for bronchial asthma in the Czech population.

Keywords: Asthma; MnSOD; Polymorphism; Superoxide dismutase

Haplotype analysis of UDP-glucuronocyltransferase 2B7 gene ( UGT2B7) polymorphisms in healthy Japanese subjects by Katsuhiko Saito; Hiroyuki Moriya; Takeru Sawaguchi; Takako Hayakawa; Seiya Nakahara; Akira Goto; Yoshiaki Arimura; Kohzoh Imai; Nahoko Kurosawa; Eiji Owada; Atsushi Miyamoto (pp. 303-308).
UDP-glucuronocyltransferase 2B7 (UGT2B7) catalyzes glucuronidation of various types of endogenous compounds and drugs, but the genetic basis of interindividual variation in the metabolism of these substances has not yet been sufficiently elucidated. In addition, information about single nucleotide polymorphisms (SNPs) and haplotypes of the UGT2B7 gene that encode the enzyme in the Japanese population is still far from sufficient.We paid special attention to and performed an investigation on −327A > G, −161T > C, −138G > A, and −125T > C in the proximal promoter region, which is regarded as being important for the transcription of the UGT2B7 gene, and also on 211G > A and 802C > T, i.e., non-synonymous SNPs of exon 1 and exon 2 that encode the substrate binding domain. Their genotypes were determined by PCR-direct sequencing.As a result of genotyping, the minor allele frequencies in 160 Japanese individuals were found to be as follows: −327SNP A allele, 0.244; −161SNP T allele, 0.244; −138SNP A allele, 0; −125SNP C allele, 0.078; 211SNP T allele, 0.148 and 802SNP T allele, 0.244. By computational haplotype analysis, it was found that these regions formed a linkage disequilibrium block, and the presence of five haplotypes was demonstrated.These results suggest that the haplotype structure in the Japanese population is different from that of other ethnic groups.

Keywords: UDP-glucuronocyltransferase 2B7; Single nucleotide polymorphism; Genotyping; Haplotype; Linkage disequilibrium

Electrochemical detection of hepatitis B surface antigen using colloidal gold nanoparticles modified by a sol–gel network interface by Dianping Tang; Ruo Yuan; Yaqin Chai; Xia Zhong; Yan Liu; Jianyuan Dai (pp. 309-314).
A novel potentiometric immunosensor for the detection of hepatitis B surface antigen has been developed by self-assembling gold nanoparticles to a thiol-containing sol–gel network.A cleaned gold electrode was first immersed in a hydrolyzed (3-mercaptopropyl) trimethoxysilane sol–gel solution to assemble a three-dimensional silica gel, and then gold nanoparticles were absorbed onto the thiol groups of the sol–gel network. Finally, hepatitis B surface antibody was assembled onto the surface of the gold nanoparticles. The self-assembling procedure was characterized by cyclic voltammetry and electrochemical impedance spectroscopy. Detection is based on the change in potentiometric response before and after the antigen–antibody reaction.Tests relating to the detection of hepatitis B surface antigen demonstrate that the potentiometric immunosensor exhibited a rapid potentiometric response (<4 min), with high sensitivity, good reproducibility, and long-term stability. The linear range was from 4 to 960 ng·mL−1 with a detection limit of 1.9 ng·mL−1 (S/N = 3) and the lifetime was 1 month.Analytical results of several specimens using the developed technique showed satisfactory agreement with those from an ELISA method. This method shows promise for detecting HBsAg in clinical specimens.

Keywords: Potentiometric immunosensor; Hepatitis B; Gold nanoparticles; Sol-gel MPS; Three-dimentional

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