Thymidylate kinase: an old topic brings new perspectives.

Thymidylate kinase (TMPK) is a key enzyme for pyrimidine synthesis that catalyzes the phosphorylation of thymidine 5'-monophosphate (dTMP) in the presence of ATP and Mg(2+) to form thymidine 5'-diphosphate (dTDP), which is then converted to thymidine 5'-triphosphate (dTTP) by nucleoside-diphosphate kinase (NDK). TMPK has an important function in cell proliferation and its enzyme kinetics and related structures have been determined in various organisms. TMPK is well recognized as a potential drug target, with the most notable function being in the activation of anti-HIV nucleoside prodrugs. Recent studies have shown that TMPK is a validated target for antibiotic development against gram-positive bacterium of Staphylococcus aureus. In addition, inhibition of human TMPK increases the potential of anticancer agent doxorubicin toward colon cancer cells regardless of p53 status. Following the rapid expanding knowledge on TMPKs and the rising interests in TMPKs as a drug target, in this review we try to describe current research on TMPKs in various organisms of eukaryotes, prokaryotes and viruses and to provide information for designing new potential inhibitors against TMPKs.

H1N1 influenza infection in children: Frequency, pattern, and outcome of chest radiographic abnormalities.

AIM: To describe the frequency, pattern, and outcome of chest radiographic abnormalities in children with H1N1 influenza infection. MATERIALS AND METHODS: Three hundred and fourteen paediatric patients with confirmed H1N1 influenza infection who underwent chest radiography at presentation at a single institution during the outbreak in 2009 were retrospectively reviewed. Abnormal chest radiographic findings related to acute infection were analysed in terms of frequency, pattern, and distribution. Medical records and follow-up radiographs were also reviewed to assess clinical features and outcomes. RESULTS: Chest lesions suggesting acute infection were identified in 49 (16%) patients (mean age 8.2 years, range approximately 1.8-18.5 years). The most common finding was prominent peribronchial marking (71%), followed by air-space opacity (51%) with or without volume decrease, generalized hyperinflation (24%), and pleural effusion (20%). Other minor findings included pneumomediastinum (n=2) and a nodule (n=1). Distributions were bilateral (55%) or unilateral (45%) with frequent involvement of lower (78%), and middle (59%) lung zones. Thirty-nine patients (80%) were hospitalized and six (12%) required mechanical ventilation, followed by recovery. Thirty-one out of the 33 patients that underwent follow-up radiography showed marked resolution of all radiographic abnormalities. CONCLUSION: The frequency of a chest radiographic abnormality was found to be low in children with H1N1 influenza infection. Although typical radiographic findings of a viral lower respiratory infection were more common, unilateral involvement and air-space opacity were common, often with pleural effusion. Furthermore, pulmonary lesions showed near complete resolution on follow-up radiographs in the majority of patients.

Population pharmacokinetics of intravenous itraconazole in patients with persistent neutropenic fever.

PURPOSE: Empirical use of intravenous (IV) itraconazole (ITZ) for febrile neutropenic patients has recently been introduced in Korea. This study was designed to investigate the population pharmacokinetics (PK) of IV-ITZ. METHODS: Sparse PK data were collected from febrile neutropenic patients undergoing empirical ITZ therapy at 200 mg/day after loading doses. NONMEM (Version. 5.1.1) was used to estimate population PK parameters. RESULTS: Forty-two patients were enrolled in the study. Mean population CL and V of IV-ITZ were 10 L/h and 1050 L, respectively. Body weight was the only contributing covariate of CL. The median simulated trough concentration of ITZ after 10 days was predicted to be about 700 ng/mL. CONCLUSIONS: In this study, we explored the population PK profile of ITZ given in IV formulation. We found that the current dosage regimen of IV-ITZ (200 mg/day) was appropriate to obtain therapeutic trough concentrations for neutropenic patients in Korea.

Infectious complications associated with alemtuzumab use for allogeneic hematopoietic stem cell transplantation: comparison with anti-thymocyte globulin.

OBJECTIVES: To evaluate the incidence of infectious complications after receiving alemtuzumab as part of a conditioning regimen for allogeneic hematopoietic stem cell transplantation (HSCT) in Korean patients. METHODS: From November 2004 to January 2006, 12 patients who received alemtuzumab-based conditioning regimens for allogeneic HSCT were evaluated retrospectively until death or until the end of the follow-up in July 2007; they were compared with 18 patients who received rabbit anti-thymocyte globulin (ATG)-containing conditioning regimens from January 2002 to January 2006. RESULTS: Post-engraftment infections occurred more frequently in the alemtuzumab recipients than in the ATG recipients; the mean number of infections, excluding cytomegalovirus (CMV) infections, per patient during the follow-up period was 2.6+/-1.4 vs. 1.0+/-0.8 (P=0.003), respectively. Although there was no statistical difference in the cumulative incidence of CMV infection between the 2 groups (91.7% vs. 55.6%, P=0.381), the alemtuzumab recipients had a higher incidence of CMV diseases (41.6% vs. 0%, P=0.0006) and a higher recurrence rate of CMV infection (90.0% vs. 27.3%, P=0.008) than did the ATG recipients, irrespective of the dose of alemtuzumab. Hemorrhagic cystitis (HC) (66.7% vs. 16.7%, P=0.009) and BK virus-associated HC (41.7% vs. 5.6%, P=0.026) developed more frequently in the alemtuzumab recipients. The all-cause mortality rate was not significantly different between the alemtuzumab and the ATG recipients (75% vs. 55.6%, P=0.28). CONCLUSION: Alemtuzumab recipients had a high incidence of CMV disease as well as BK virus-associated HC compared with the ATG recipients. The dose of alemtuzumab should be tailored to patients' risk; in addition, the implementation of the appropriate prophylaxis for CMV and early detection strategies for BK virus are recommended.

Inhibition of UL54 and UL97 genes of human cytomegalovirus by RNA interference.

Short interfering RNAs (siRNAs), namely siUL54-1 and siU54-2 targeting UL54 (DNA polymerase) gene, and siUL97-1 and siUL97-2 targeting UL97 (phosphotransferase) gene, were used to inhibit respective genes of Human cytomegalovirus (HCMV) and consequently the virus infection process in human foreskin fibroblast (HFF) cultures. The virus infection was monitored by cell morphology (CPE), levels of UL83 and IE86 mRNAs, and virus antigen. The results showed that siUL97-2 remarkably inhibited viral CPE while other siRNAs were less inhibitory. The siRNAs reduced the levels of UL83 mRNA but not that of IE86 mRNA; again, siUL97-2 was most inhibitory. Particularly, siUL97-2 reduced the UL83 mRNA level 14, 19, 203, and 37 times at 24, 48, 72, and 96 hrs post infection (p.i.), respectively. When tested for the effect on viral antigen by immunofluorescent assay (IFA), UL97-2 exerted a marked inhibition. These results demonstrate the effectiveness of siRNAs against experimental HCMV infection and indicate their therapeutic potential.

Application of a combined process of moving-bed biofilm reactor (MBBR) and chemical coagulation for dyeing wastewater treatment.

A combined process consisted of a Moving-Bed Biofilm Reactor (MBBR) and chemical coagulation was investigated for textile wastewater treatment. The pilot scale MBBR system is composed of three MBBRs (anaerobic, aerobic-1 and aerobic-2 in series), each reactor was filled with 20% (v/v) of polyurethane-activated carbon (PU-AC) carrier for biological treatment followed by chemical coagulation with FeCl2. ln the MBBR process, 85% of COD and 70% of color (influent COD = 807.5 mg/L and color = 3,400 PtCo unit) were removed using relatively low MLSS concentration and short hydraulic retention time (HRT = 44 hr). The biologically treated dyeing wastewater was subjected to chemical coagulation. After coagulation with FeCl2, 95% of COD and 97% of color were removed overall. The combined process of MBBR and chemical coagulation has promising potential for dyeing wastewater treatment.

Infectious complications and outcomes after allogeneic hematopoietic stem cell transplantation in Korea.

We reviewed 242 allogeneic hematopoietic stem cell transplantation (HSCT) recipients retrospectively over a 2-year period (January 1998-December 1999) in order to analyze the characteristics and assess the outcomes of infectious complications in patients after HSCT in Korea. Bacteria were the major pathogens before engraftment, and viral and fungal infections predominated during the post-engraftment period. Varicella zoster virus was the most common viral pathogen after engraftment. Cytomegalovirus disease occurred mainly in the late-recovery phase. The frequency of mold infection was higher than that of yeast. There was a relatively high incidence of tuberculosis (3.0%) and Pneumocystis carinii pneumonia (6.5%). One case of death by measles confirmed by autopsy was also noted. Overall, cumulative mortality was 43% (104/242), and 59.6% of these deaths (62/104) were infection-related. Allogeneic HSCT recipients from unrelated donors were prone to infectious complication and higher mortality than those from matched sibling (17/39 (43.6%) vs 45/203 (22.2%), respectively; P<0.01; odd ratio 2.5; 95% confidence interval 1.2-5.1). As infection was the main post-HSCT complication in our data, more attention should be given to the management of infections in HSCT recipients.

High fidelity SNP genotyping using sequence-specific primer elongation and fluorescence correlation spectroscopy.

Reliable, efficient and cost-effective modalities are urgently needed for mass screening of gene mutations. Previous reports have shown that SSCP or genechip methods require substantial time and monetary costs, thus limiting their appeal. Sequence Specific Primer Polymerase Chain Reaction (SSP-PCR) is a reliable and cost-effective method that utilizes the 3'-end discrimination properties of polymerase. However, the applicability of conventional SSP-PCR is limited due to the difficulties associated with determining optimal conditions and because mis-matched primers are amplified, resulting in signal noise during end-point assay. To overcome this problem, we eliminated the reverse primers from SSP-PCR, thus preventing amplification of mis-matched primers. We designated this method Sequence-Specific Primer Cycle Elongation (SSPCE). However, the detection of elongated sequence specific primers was difficult using conventional electrophoresis due to the small amounts of amplification product present. We therefore combined SSPCE and Fluorescence Correlation Spectroscopy, which is a novel technique used to determine the number and size of fluorophores at nano-molar concentrations, and designated the method SSPCE-FCS. We compared conventional SSP-PCR and SSPCE-FCS with regard to determining optimal conditions using two Mitochondrial SNPs (G --> A at position 1598, G --> A at position 12192). We were able to determine the optimal conditions for the SNP at position 1598 using either method. However, optimal conditions could only be determined for SSPCE-FCS with the 12192 mutation because non-specific amplification was observed at a wide range of annealing temperatures in SSP-PCR. We then applied this method to three other SNPs and the results were consistent with the results of sequencing data.

Oxygen dynamics in petroleum hydrocarbon contaminated salt marsh soils: III. A rate model.

A model describing oxygen dynamics due to crude oil biodegradation under flooded conditions in saltwater wetlands was developed. The model is composed of three non-linear ordinary differential equations (ODEs) that simulate oxygen uptake, cell growth, and oil degradation simultaneously. The model equations were solved by using a stiff version of ODE solver, ODEPACK, which employs a multistep method and allows the change of step sizes and order of methods (ie., Gear's method). The results of model simulation were compared with experimental data obtained from a fully aerated microcosm study. The results of model simulation indicate that dissolved oxygen concentration in the overlying water rapidly depleted below 3 mg l(-1) unless the reaeration coefficient was higher than 2.0 day(-1). Active aerobic biodegradation of crude oil did not occur under flooded conditions because (i) dissolved oxygen is rapidly depleted, (ii) reaeration is not sufficient enough to replenish dissolved oxygen and (iii) the oil dissolution rate constant decreases over time. The model may lead to better understanding of oxygen demand for a long bioremediation period. The results of this study may be applicable for the establishment of an engineered bioremediation strategy.

Sequential sorption and desorption of chlorinated phenols in organoclays.

Effect of pH on the sorption and desorption of the chlorinated phenols (2-chlorophenol and 2,4-dichlorophenol) in HDTMA-montmorillonite organoclays was investigated using sequential batch experiments. 2,4-dichlorophenol exhibited higher affinity in both sorption and desorption than 2-chlorophenol at pH 4.85 and 9.15. For both chlorophenols, the protonated speciation (at pH 4.85) exhibited a higher affinity in both sorption and desorption than the predominant deprotonated speciation (about 80% and 95% of 2-chlorophenate and 2,4-dichlophenate anions at pH 9.15, respectively). Desorption of chlorinated phenols was strongly dependent on the current pH regardless of their speciation during the previous sorption stage. No appreciable desorption resistance of the chlorinated phenols was observed in organoclays after sequential desorptions. Affinity of both chlorophenols in bisolute competitive sorption and desorption was reduced compared to that in a single-solute system due to the competition between solutes. The ideal adsorbed solution theory coupled with the single-solute Freundlich model successfully predicted the bisolute competitive sorption and desorption equilibria.

Benefits-based analysis of visitor use of Sorak-san National Park in Korea.

Benefits-based management seeks to provide recreation benefits for recreation participants by managing the physical environments in which recreation occurs. This study investigates the relationship between benefits desired by visitors and the physical, social, and managerial characteristics of settings that facilitate realization of recreation benefits. Data such as perceived benefits from recreation experiences, setting attributes that significantly influenced perceived benefits, and sociodemographic variables were collected from 376 visitors to Sorak-san National Park in the eastern part of the Korean peninsula. Cluster analysis was used to group visitors' desired benefits into 12 domains: relationship with nature/scenery, escaping pressure, learning about nature, family togetherness, introspection, exploration, autonomy/achievement, being with friends, leading others, skills/learning, risk taking, and meeting/observing new people. Multiple regression was used to link benefit domains with the characteristics of settings. The social attribute of recreation settings was linked to eight of the ten benefit domains. There were two statistically strong multiple regression correlations: (1) between domain of "relationship with nature/scenery" and the attributes "forest/water," "attractive nature," and "facility/maintenance" and (2) between the domain of "escaping pressure" and the attributes "attractive nature" and "social." The results of this study are useful to managers in their efforts to provide recreation opportunities for visitors to achieve beneficial outcomes.

Improvement of water treatment performance by using polyamine flocculants.

In this study, polyamine flocculants were synthesized and applied to Nak-dong river raw water in Korea to examine their efficiency in reducing turbidity, total organic carbon (TOC) and UV254. Synthesized polyamines were effective as flocculants for water treatment and the addition of organic polymer caused a reduction of 50-80% of the consumption of polyaluminium chloride (PAC). The effects of polyamine on the removal of turbidity, TOC and UV254 were investigated via both jar and pilot tests. The adsorption and separation mechanisms for the removal of turbidity and TOC by using the polymer flocculants were also observed.

Oxygen dynamics in crude oil contaminated salt marshes: I. Aerobic respiration model.

A simple model was developed to predict oxygen demand exerted by aerobic biodegradation of spilled crude oil and fertilizer added to stimulate biodegradation in salt marsh sediment. The role of aerobic respiration (AR) was determined using first-order G kinetics. The G kinetic rate constants were calculated from laboratory data sets through linear regression. The effect of oil and fertilizer on AR was quantified by comparing three treatments: (i) control (no oil and fertilizer amended), (ii) fertilized, and (iii) oiled and fertilized sediments. The effects of dissolved oxygen concentration in the overlying water and surface mass transfer coefficient were investigated. Aerobic respiration was strongly dependent on the overlying dissolved oxygen concentration when crude oil and fertilizer were added. Oiled/fertilized cores did not show higher SOD and AR than control cores when overlying DO concentration dropped below 5 mg l(-1). Results indicate that higher aerobic respiration in oiled/fertilized is exerted by aerobic biodegradation of crude oil and that major biodegradation of crude oil occurs only when DO level is high (> 5 mg l(-1)).

Oxygen dynamics in crude oil contaminated salt marshes: II. Carbonaceous sediment oxygen demand model.

The role of carbonaceous sediment oxygen demand (CSOD) due to oxidation of sulfides reduced in oxygen dynamics in crude oil contaminated salt marshes was investigated through a mathematical model. An existing CSOD model was calibrated from laboratory data sets through nonlinear regression. The effect of oil and fertilizer on CSOD was quantified by comparing three treatments (i) control (no oil and fertilizer amended) (ii) fertilized, and (iii) oiled and fertilized sediments. CSOD was directly proportional to sulfate reduction. Higher CSODs under oiled/fertilized and fertilized conditions were primarily due to increased sulfate reduction rates under these conditions. Reaction velocity for oxidation of dissolved sulfide in the aerobic layer, kappaH2S,d1, was significantly greater than that of particulate sulfide oxidation, kappaH2S,p1. This indicates that dissolved sulfides are dominant over particulate sulfides and directly related to CSOD in salt marshes. The CSOD was linearly dependent on the overlying dissolved oxygen concentration when crude oil and fertilizer were added.

Morphological and physiological restorations of hereditary form of dilated cardiomyopathy by somatic gene therapy.

TO-2 strain hamsters with dilated cardiomyopathy, gene deletion of delta-sarcoglycan (SG) and no expression of alpha-, beta-, gamma-, and delta-SG proteins are useful for developing the potential gene therapy of intractable heart failure. We prepared recombinant adeno-associated virus vector including normal delta-SG gene driven by CMV promoter and intramurally administered in vivo. The transfected myocardium induced robust expression of both transcript and transgene for 2/3 period of the animal's life expectancy. Immunostaining demonstrated reexpression of not only delta-SG but also other three SGs in 40% cells in the transfected region and normalization of the diameter of transduced cardiomyocytes. Hemodynamic study revealed preferential amelioration of the diastolic indices (LVEDP, the dP/dt(min) and CVP). These results provide the first evidence that supplementation of a specific gene with efficient and sustained transfection capability restores the genetic, morphological, and functional deteriorations.

Hepatosplenic tuberculosis mimicking disseminated candidiasis in patients with acute leukemia.

Two cases of hepatosplenic tuberculosis in patients with acute leukemia during or after chemotherapy following prolonged neutropenia are presented. Tuberculosis should be considered as one cause of hepatosplenic abscesses during prolonged neutropenia, especially in countries where the disease is endemic.