The usefulness of non-invasive co-oximetry haemoglobin measurement for screening pre-operative anaemia.

Pre-operative anaemia (haemoglobin < 13.0 g.dl-1 ) is a modifiable peri-operative risk-factor. This is screened for using formal laboratory testing. A non-invasive finger-probe sensor that can accurately measure haemoglobin is a possible alternative. This study considers the accuracy of non-invasive haemoglobin measurement using the Rad-67™ Rainbow (Masimo Corp., Irvine, CA, USA) compared with formal laboratory testing and its usefulness in detecting pre-operative anaemia. A total of 392 patients had measurements taken for non-invasive haemoglobin and perfusion index values using the Rad-67 Rainbow, alongside further peri-operative parameters and a formal laboratory haemoglobin test. Bland-Altman and sensitivity analysis showed that the limits of agreement between non-invasive and formal laboratory haemoglobin testing were between -1.95 g.dl-1 and 2.23 g.dl-1 (p < 0.001). The overall performance of non-invasive haemoglobin measurement was better in men than women (ROC 91.1% vs. 78.2%) and less biased in men, mean -0.08 (SD 1.09, 95%Cl -0.23-0.07) compared with women (mean 0.38 (SD 0.99, 95%CI 0.24-0.52)). Pre-operative anaemia was more prevalent in women than men (50.3% vs. 14.4%). The sensitivity of non-invasive anaemia detection (haemoglobin < 13 g.dl-1 ) was 66% for women and 52% for men. A non-invasive haemoglobin value of 14.0 g.dl-1 had an overall 91% sensitivity for detecting pre-operative anaemia (82% in men and 93% in women). The Rad-67 Rainbow is inadequate for the estimation of formal laboratory haemoglobin and lacks sensitivity for detecting pre-operative anaemia, especially in women. Further advancement in technology and accuracy is needed before it can be recommended as a routine pre-operative screening test.

Characteristics of color optical shutter with dye-doped polymer network liquid crystal.

The optical properties and the theoretical prediction of color optical shutter with dye-doped polymer network liquid crystal (PNLC) were investigated. The view-angle dependence of reflectance according to the bias conditions showed distinctive characteristics, which could be explained from the effects of dye absorption and path length. It was also shown that the thickness dependence of reflectance was strongly influenced by the light-scattering coefficient. Our experimental results matched up well with the theoretical prediction based on the light scattering of liquid crystals in polymer network and the absorption of dichroic dye. This work indicates potential to improve the optical device using dye-doped liquid crystal-polymer composite.

Removal of ammonium ion from aqueous solution using magnetically modified zeolite.

This paper assesses the potential of magnetically modifiedzeolite (MMZ) for ammonia removal from aqueous solution. The effects of relevant parameters such as contact time, pH and initial ammonia concentration were examined. The results show that ammonium ion removal by MMZ occurs rapidly within the first five minutes, pH has an effect on ammonium ion removal efficiency, and the ammonium ion removal capacity of MMZ increases with increase in initial ammonium ion concentration. The Langmuir and Freundlich adsorption isotherms were used to determine the adsorption characteristics. Ammonium-laden MMZ seems to be regenerated completely by sodium chloride solution at pH 11-12.

Sequence-based screening for self-sufficient P450 monooxygenase from a metagenome library.

AIMS: Cytochrome P450 monooxygenases (CYPs) are useful catalysts for oxidation reactions. Self-sufficient CYPs harbour a reductive domain covalently connected to a P450 domain and are known for their robust catalytic activity with great potential as biocatalysts. In an effort to expand genetic sources of self-sufficient CYPs, we devised a sequence-based screening system to identify them in a soil metagenome. METHODS AND RESULTS: We constructed a soil metagenome library and performed sequence-based screening for self-sufficient CYP genes. A new CYP gene, syk181, was identified from the metagenome library. Phylogenetic analysis revealed that SYK181 formed a distinct phylogenic line with 46% amino-acid-sequence identity to CYP102A1 which has been extensively studied as a fatty acid hydroxylase. The heterologously expressed SYK181 showed significant hydroxylase activity towards naphthalene and phenanthrene as well as towards fatty acids. CONCLUSIONS: Sequence-based screening of metagenome libraries is expected to be a useful approach for searching self-sufficient CYP genes. The translated product of syk181 shows self-sufficient hydroxylase activity towards fatty acids and aromatic compounds. SIGNIFICANCE AND IMPACT OF THE STUDY: SYK181 is the first self-sufficient CYP obtained directly from a metagenome library. The genetic and biochemical information on SYK181 are expected to be helpful for engineering self-sufficient CYPs with broader catalytic activities towards various substrates, which would be useful for bioconversion of natural products and biodegradation of organic chemicals.

Evaluation of endoscopic hemostasis in upper gastrointestinal bleeding related to Mallory-Weiss syndrome.

BACKGROUND AND STUDY AIMS: The endoscopic hemostatic method has been introduced as a safe and effective mechanical approach to hemostasis for upper gastrointestinal bleeding related to Mallory-Weiss syndrome (MWS). However, the indications for when to use endoscopic treatment are debatable because many patients need only medical observation. The study was designed to evaluate the necessity and efficacy of endoscopic hemostasis in upper gastrointestinal bleeding related to MWS. PATIENTS AND METHODS: From July 1994 to May 2000, we conducted a clinical trial in 76 patients who were found by endoscopy to have active bleeding (I, spurting; II, oozing), protruding visible vessels (III), and/or adherent clots (IV). Two study periods can be differentiated: in the first 3 years endoscopic treatment (n = 30) was prospectively analyzed and in the final 3 years medical treatment (n = 46) was analyzed in both cases to compare the outcome in MWS bleeding II-IV. In the first study period, in addition, endoscopic treatment was randomised to an injection method, using a mixture of hypertonic saline and epinephrine (HSE) (n = 14) and a hemoclipping or band ligation method (n = 16). RESULTS: Rebleeding was observed in four of 14 patients who had received endoscopic hemostasis with HSE injection and one of 46 patients who had been managed with medical treatment. No rebleeding was found following hemoclipping or band ligation. While all rebleeding was in bleeding stigmata of the I (1) and II (4) grades, there was no rebleeding in protruding visible vessels (III) or in adherent clots (IV), regardless of treatment methods. CONCLUSIONS: Our results suggested that endoscopic hemostasis is not necessary in patients without active bleeding stigmata, and the mechanical hemostatic method is more effective than HSE injection in patients with active bleeding stigmata.

Is tumor necrosis factor genotype (TNFA2/TNFA2)a genetic prognostic factor of an unfavorable outcome in IgA nephropathy?

The purpose of this study was to examine whether there are the associations between TNF alpha and TNF beta gene polymorphisms and the development and progression of Ig A nephropathy (IgAN). A cross-sectional study on TNF alpha and TNF betagene polymorphisms by polymerase chain reaction with restriction fragment length poly-morphisms was performed on 76 patients with primary IgAN confirmed by renal biopsy and 100 healthy controls. The allele with G-->A substitution was designated as TNFA2 for the TNF alpha gene and TNFB2 for the TNF betagene. A patient in whom dialysis treatment was started or whose serum creatinine became double or over during the follow-up duration was designated as a "progressor". The TNFA2/ TNFA2 genotype was more prevalent in the progressor than in the non-pregressor group (20.0 vs 3.3%, p<0.05). Clinical factors such as serum creatinine, systolic and diastolic blood pressure (p<0.001, respectively) were higher and pathologic factor such as Grade IV or V renal lesions was more prevalent (p<0.01) in the progressor than in the non-progressor group. Therefore, TNFA2/TNFA2 genotype may be a risk factor for the progression of IgAN.

Endoscopic factors predisposing to rebleeding following endoscopic hemostasis in bleeding peptic ulcers.

BACKGROUND AND STUDY AIMS: Various clinical and endoscopic factors have been proposed and used as predictors of endoscopic treatment failure in bleeding peptic ulcers. Recently, several endoscopic factors have been considered to be more significant than various clinical factors, except for shock. Detailed knowledge of which endoscopic factors can be classified as predictors of rebleeding following endoscopic hemostasis is needed. This study describes newly defined endoscopic variables and evaluates their usefulness as predictors of endoscopic treatment failure. PATIENTS AND METHODS: Between January 1996 and April 1999, diagnostic and therapeutic endoscopies were carried out in 143 patients with active bleeding peptic ulcers. Nine clinical and eight endoscopic variables were studied. Endoscopic factors were classified by three types of stigmata bleeding, 14 locations, two ulcer sizes, three ulcer bases, three visible vessel colors, two ulcer depths, two margin shapes, and three endoscopic treatment methods (injection, hemoclipping, and combination). RESULTS: 36 patients experienced rebleeding (25.2 %), 11 patients needed operations (7.7 %) and five deaths occurred (3.5 %). In univariate analysis, rebleeding was significantly related to: i) presence of spurting activity (odds ratio [OR] = 4.91, P = 0.006), ii) ulcer size larger than 2 cm (OR = 2.78, P = 0.017); and iii) location in stomach (OR = 2.81, P = 0.026). Clinical variables including age, shock, and initial hemoglobin levels were not significantly related to rebleeding. In multiple logistic regression using selected significant variables, presence of spurting activity remained a significant independent predictor of rebleeding (adjusted OR = 6.48, P = 0.003). CONCLUSION: Our data support the hypothesis that endoscopic factors are more important than clinical ones when predicting rebleeding of peptic ulcers. Based on statistical analysis of risk factors, the ulcers most likely to rebleed after endoscopic therapy are those which are located in the stomach, are greater than 2 cm in diameter and exhibit oozing or spurting of blood.

Mutational effects on thermostable superoxide dismutase from Aquifex pyrophilus: understanding the molecular basis of protein thermostability.

We designed two mutants of superoxide dismutase (SOD), one is thermostable and the other is thermolabile, which provide valuable insight to identify amino acid residues essential for the thermostability of the SOD from Aquifex pyrophilus (ApSOD). The mutant K12A, in which Lys12 was replaced by Ala, had increased thermostability compared to that of the wild type. The T(1/2) value of K12A was 210 min and that of the wild type was 175 min at 95 degrees C. However, the thermostability of the mutant E41A, which has a T(1/2) value of 25 min at 95 degrees C, was significantly decreased compared to the wild type of ApSOD. To explain the enhanced thermostability of K12A and thermolabile E41A on the structural basis, the crystal structures of the two SOD mutants have been determined. The results have clearly shown the general significance of hydrogen bonds and ion-pair network in the thermostability of proteins.

Prevalence of arthropod antibodies in Korean patients with allergic rhinitis.

Arthropod antigens are main causative agents which induce allergic responses in humans. However, little information is known about the prevalence of specific arthropod allergens in Koreans with allergic diseases. The current study was designed to determine the positive rates of arthropod antibodies by the Korean inhalant panel of MAST-CLA. One hundred sixty patients, who were diagnosed with allergic rhinitis from an out-patient center at the Soonchunhyang University Chunan Hospital, were studied between August 1998 to July 2000. The overall positive rate, at least more than one specific antibody of arthropods such as Dermatophagoides farinae (Df), Dermatophagoides pteronyssinus (Dp), and cockroach mix (Cm), was 46.9%. Each positive rate of Df, Dp, and Cm was 45.0%, 43.1%, and 8.8%, respectively. A significant agreement among arthropod allergens was observed (Df and Dp: 95.6%, Kappa = 0.911, P < 0.001). Our data supported the fact that arthropods were the most common allergens in Korean patients with allergic rhinitis; however, the MAST-CLA should be modified to increase specificity of arthropod allergens.

Associations of lead exposure and dose measures with erythrocyte protein kinase C activity in 212 current Korean lead workers.

Lead can replace calcium in enzyme assays that measure protein kinase C activity and lead activates protein kinase C in human erythrocytes after exposure to lead in vitro. To examine the relevance of these observations to lead exposure in humans, we studied the associations of lead found in blood or tibia with activation of protein kinase C in erythrocytes isolated from workers in the lead industry. We examined erythrocytes among 212 lead workers, with a mean (+/-SD) age of 39.1 (10.0) years and exposure duration of 8.1 (6.5) years and measured protein kinase C activation by an in vitro back-phosphorylation assay. After adjustment for potential confounding factors (age and sex), tibia lead and exposure duration were significantly associated with erythrocyte protein kinase C activation (both p values < 0.05). No associations were observed between protein kinase C activation and blood-lead or zinc-protoporphyrin levels. These findings suggest that human exposure to lead results in activation of erythrocyte protein kinase C, which may be directly relevant to the neurotoxicity of lead.

4G/5G polymorphism of the plasminogen activator inhibitor-1 gene and insertion/deletion polymorphism of the tissue-type plasminogen activator gene in atherothrombotic stroke.

BACKGROUND AND PURPOSE: Decreased fibrinolytic capacity due to increased plasminogen activator inhibitor-1 (PAI-1) activity and decreased tissue-type plasminogen activator (t-PA) activity has been associated with hypertension or atherothrombotic disorders. The aims of this study were to observe associations of the genetic polymorphism for PAI-1 and t-PA with hypertension and atherothrombotic stroke, and to elucidate whether impaired fibrinolytic activity in atherothrombotic stroke was related to atherothrombosis per se or to other risk factors such as hypertension. METHODS: Patients with atherothrombotic stroke (n = 60), hypertension (n = 100), and control subjects (n = 100) were enrolled. We genotyped all subjects for 4G/5G polymorphism in the promoter region of the PAI-1 gene and the Alu-repeat insertion/deletion (I/D) polymorphism in intron h of the t-PA gene by polymerase chain reaction and endonuclease digestion. RESULTS: The frequency of the 4G/4G genotype of PAI-1 was significantly higher in the atherothrombotic stroke patients than the control subjects (41.7 versus 21%; p = 0.005), but not in the hypertensive subjects. There was a significant association between 4G/4G genotype of PAI-1 and atherothrombotic stroke (adjusted odds ratio = 3.11, 95% confidence interval 1.18-8.15), adjusting for age, sex, total cholesterol, low-density lipoprotein, triglyceride, and body mass index. However, the number of the I/I genotype of t-PA in the atherothrombotic stroke or hypertensive patients was virtually identical to the control subjects. CONCLUSION: Our results suggest that the 4G/4G genotype of the PAI-1 gene is significantly associated with an increased risk of atherothrombotic stroke. This finding also supports that impaired fibrinolytic activity in atherothrombotic stroke is related to atherothrombosis per se, but not to hypertension, one of the most important risk factors of atherothrombotic stroke.

The methylenetetrahydrofolate reductase gene polymorphism in Koreans with coronary artery disease.

Hyperhomocysteinemia is a known risk factor of cardiovascular diseases. Methylenetetrahydrofolate reductase (MTHFR), involved in folate-dependant metabolism, is associated with homocysteine levels. We studied the associations among MTHFR genotypes, coronary artery disease (CAD), and homocysteine levels in 85 patients with CAD and 152 healthy subjects. The MTHFR genotypes and plasma homocysteine levels were determined. No significant difference in mutation of the MTHFR gene between two groups was observed (P>0.05). While the homozygous mutant genotype (V/V) had the highest homocysteine levels compared to wild (A/A) and heterozygous mutant (A/V) genotypes, there were no significant differences in homocysteine levels among the MTHFR genotype groups. Homocysteine was significantly and inversely related to folate levels, the significant association in V/V genotype (beta coefficient=-1.954, P=0.04). Our data suggested that MTHFR polymorphism was not associated with homocysteine levels, implying no association between gene polymorphism and CAD in Koreans.

Associations of blood lead, dimercaptosuccinic acid-chelatable lead, and tibia lead with neurobehavioral test scores in South Korean lead workers.

The authors performed a cross-sectional study to evaluate associations between blood lead, tibia lead, and dimercaptosuccinic acid (DMSA)-chelatable lead and measures of neurobehavioral and peripheral nervous system function among 803 lead-exposed workers and 135 unexposed controls in South Korea. The workers and controls were enrolled in the study between October 1997 and August 1999. Central nervous system function was assessed with a modified version of the World Health Organization Neurobehavioral Core Test Battery. Peripheral nervous system function was assessed by measuring pinch and grip strength and peripheral vibration thresholds. After adjustment for covariates, the signs of the beta coefficients for blood lead were negative for 16 of the 19 tests and blood lead was a significant predictor of worse performance on eight tests. On average, for the eight tests that were significantly associated with blood lead levels, an increase in blood lead of 5 microg/dl was equivalent to an increase of 1.05 years in age. In contrast, after adjustment for covariates, tibia lead level was not associated with neurobehavioral test scores. Associations with DMSA-chelatable lead were similar to those for blood lead. In these currently exposed workers, blood lead was a better predictor of neurobehavioral performance than was tibia or DMSA-chelatable lead, mainly in the domains of executive abilities, manual dexterity, and peripheral motor strength.

Rapid reductive destruction of hazardous organic compounds by nanoscale Fe0.

Fe0-mediated reductive destruction of hazardous organic compounds such as chlorinated organic compounds (COCs) and nitroaromatic compounds (NACs) in the aqueous phase is one of the latest innovative technologies. In this paper, rapid reductive degradation of COCs and NACs by synthesized nanoscale Fe0 in anaerobic batch systems was presented. The nanoscale Fe0, characterized by high specific surface area and high reactivity, rapidly transformed trichloroethylene (TCE), chloroform (CF), nitrobenzene (NB), nitrotoluene (NT), dinitrobenzene (DNB) and dinitrotoluene (DNT) under ambient conditions, which results in complete disappearance of the parent compounds from the aqueous phase within a few minutes. GC analysis reported that the main products of the dechlorination of TCE and CF were ethane and methane as well as that most of the nitro groups in NACs were reductively transformed to amine groups. These results suggest that the rapid reductive destruction by nanoscale Fe0 is potentially a viable in situ or aboveground treatment of groundwater contaminated with hazardous organic compounds including COCs and NACs.

Kinetics of reductive denitrification by nanoscale zero-valent iron.

Zero-valent iron powder (Fe0) has been determined to be potentially useful for the removal of nitrate in the water environment. This research is aimed at subjecting the kinetics of denitrification by nanoscale Fe0 to an analysis of factors affecting the chemical denitrification of nitrate. Nanoscale iron particles with a diameter in the range of 1-100 nm, which are characterized by the large BET specific surface area to mass ratio (31.4 m2/g), removed mostly 50, 100, 200, and 400 mg/l of nitrate within a period of 30 min with little intermediates. Compared with microscale (75-150 microm) Fe0, end product is not ammonia but N2 gas. Kinetics analysis from batch studies revealed that the denitrification reaction with nanoscale Fe0 appeared to be a pseudo first-order with respect to substrate and the observed reaction rate constant (k(obs)) varied with iron content at a relatively low degree of application. The effects of mixing intensity (rpm) on the denitrification rate suggest that the denitrification appears to be coupled with oxidative dissolution of iron through a largely mass transport-limited surface reaction (<40 rpm).

Crystallization and preliminary x-ray crystallographic analysis of NAD+-dependent DNA ligase from Thermus filiformis.

A highly thermostable DNA ligase from Thermus filiformis has been crystallized at room temperature using methoxypolyethylene glycol 5000 as a precipitant. The crystal belongs to the monoclinic space group P2(1), with unit-cell parameters a = 90.63, b = 117.80, c = 98. 65 A, beta = 115.56 degrees. Two molecules of DNA ligase are present in the asymmetric unit, giving a crystal volume per protein mass (V(m)) of 3.1 A(3) Da(-1) and a solvent content of 61%. A native data set extending to 3.0 A resolution has been collected at 100 K using synchrotron X-rays.

Associations between laboratory parameters and outcome of paraquat poisoning.

Paraquat, a non-selective herbicide, is a known fatal substance in humans, and intentional ingestion of paraquat is increasing among Korean suicides. In 1999, 147 subjects admitted to the Institute of Pesticide Poisoning, Soonchunhyang Chunan Hospital, Korea ingested paraquat. Initial routine laboratory tests were conducted and the outcome of paraquat poisoning was categorized as survivor and fatality. Mean amount (S.D.) of ingestion was 54.5 (104.9) ml, and the overall fatality rate was 44.2%. Abnormal liver function (GOT and GPT), renal dysfunction (BUN and creatinine), metabolic acidosis (pH and PaCO(2)), and abnormal urine analysis (RBC, WBC, and protein) had significant odds ratios (ORs) for paraquat fatality (P<0.05). In multiple logistic regression, subjects with liver or renal dysfunction or metabolic acidosis had significant risks of the fatality. Our results determined that initial routine laboratory parameters could be used to predict the outcome of paraquat poisoning and recommended that evaluation of acid-base status and renal and liver function should be conducted and evaluated before intensive therapy.

Structure-based identification of a novel NTPase from Methanococcus jannaschii.

Almost half of the entire set of predicted genomic products from Methanococcus jannaschii are classified as functionally unknown hypothetical proteins. We present a structure-based identification of the biochemical function of a protein with an as yet unknown function from a M. jannaschii gene, Mj0226. The crystal structure of Mj0226 protein determined at 2.2 A resolution reveals that the protein is a homodimer and each monomer folds into an elongated alpha/beta structure of a new fold family. Comparisons of Mj0226 protein with protein structures in the database, however, indicate that one part of the protein is homologous to some of the nucleotide-binding proteins. Biochemical analysis shows that Mj0226 protein is a novel nucleotide triphosphatase that can efficiently hydrolyze nonstandard nucleotides such as XTP to XMP or ITP to IMP, but not the standard nucleotides, in the presence of Mg2+ or Mn2+ ions.

Structure and mechanism of glutamate racemase from Aquifex pyrophilus.

Glutamate racemase (MurI) is responsible for the synthesis of D-glutamate, an essential building block of the peptidoglycan layer in bacterial cell walls. The crystal structure of glutamate racemase from Aquifex pyrophilus, determined at 2.3 A resolution, reveals that the enzyme forms a dimer and each monomer consists of two alpha/beta fold domains, a unique structure that has not been observed in other racemases or members of an enolase superfamily. A substrate analog, D-glutamine, binds to the deep pocket formed by conserved residues from two monomers. The structural and mutational analyses allow us to propose a mechanism of metal cofactor-independent glutamate racemase in which two cysteine residues are involved in catalysis.

Structural basis for cold adaptation. Sequence, biochemical properties, and crystal structure of malate dehydrogenase from a psychrophile Aquaspirillium arcticum.

Aquaspillium arcticum is a psychrophilic bacterium that was isolated from arctic sediment and grows optimally at 4 degrees C. We have cloned, purified, and characterized malate dehydrogenase from A. arcticum (Aa MDH). We also have determined the crystal structures of apo-Aa MDH, Aa MDH.NADH binary complex, and Aa MDH.NAD.oxaloacetate ternary complex at 1.9-, 2.1-, and 2.5-A resolutions, respectively. The Aa MDH sequence is most closely related to the sequence of a thermophilic MDH from Thermus flavus (Tf MDH), showing 61% sequence identity and over 90% sequence similarity. Stability studies show that Aa MDH has a half-life of 10 min at 55 degrees C, whereas Tf MDH is fully active at 90 degrees C for 1 h. Aa MDH shows 2-3-fold higher catalytic efficiency compared with a mesophilic or a thermophilic MDH at the temperature range 4-10 degrees C. Structural comparison of Aa MDH and Tf MDH suggests that the increased relative flexibility of active site residues, favorable surface charge distribution for substrate and cofactor, and the reduced intersubunit ion pair interactions may be the major factors for the efficient catalytic activity of Aa MDH at low temperatures.