Changes in blood and urinary cadmium levels and bone mineral density according to osteoporosis medication in individuals with an increased cadmium body burden.

The aim of this study was to assess changes in bone mineral density (BMD) and cadmium (Cd) levels in blood and urine in individuals living in a Cd-contaminated area according to the type of osteoporosis medication over a three-year period. This follow-up study included 204 residents living in the vicinity of a closed copper refinery, who had been found to have elevated urinary or blood Cd levels. Cd levels in the blood and urine, as well as BMD, were measured every 6 months. After the first BMD measurement, individuals were prescribed antiresorptives such as alendronate or vitamin D and calcium, according to their BMD. Subjects were classified according to the type of medicine provided over the previous 6 months. General linear models controlling for other factors were used to evaluate the effects of each type of medication on the participants' Cd levels and BMD. Spinal BMD showed a significant increase in the antiresorptive group compared to the nontreatment group. Significant decreases in blood Cd levels were found in the vitamin D and calcium group, in comparison to the nontreatment group, as well as a marginally significant decrease in the antiresorptive group. The vitamin D and calcium group showed a significantly greater decrease in urinary Cd levels than the nontreatment group. In contrast, antiresorptive medication was found to have a negative effect on urinary Cd excretion. These results suggest that vitamin D and calcium treatment for osteoporosis lowers blood Cd levels more effectively and improves urinary Cd excretion.

Impact of surgical pulmonary valve replacement on ventricular mechanics in children with repaired tetralogy of Fallot.

Impaired ventricular myocardial mechanics are observed in patients with repaired tetralogy of Fallot (rTOF). Effects of pulmonary valve replacement (PVR) on ventricular remodeling are controversial. The objective was to assess the impact of surgical PVR on ventricular mechanics in pediatric patients after rTOF. Speckle-tracking analysis was performed in 50 rTOF children, aged 12.6 ± 3.3 years, pre-operatively and 14.5 ± 2.2 months post-PVR. Early post-operative studies 2.2 ± 0.6 months post-PVR were performed in 28 patients. Cardiac magnetic resonance (CMR) pre- and post-PVR was collected. Mid-term post-PVR right ventricular (RV) longitudinal strain increased above pre-operative strain (-19.2 ± 2.7 to -22.0 ± 3.0%, p < 0.001) with increases observed in individual RV segments. Left ventricular (LV) strain did not differ at medium-term follow-up. LV and RV longitudinal strain was reduced early post-operatively, followed by recovery of biventricular systolic strain by mid-term follow-up. CMR RV end-diastolic indexed volumes correlated with RV strain pre-operatively (r = 0.432, p = 0.005) and at mid-term follow-up (r = 0.532, p = 0.001). Volume-loaded RVs had reduced early RV basal longitudinal strain compared to pressure-loading conditions. Reversed basal counterclockwise rotation was associated with lower mid-term global LV and basal RV strain compared to patients with normal rotation. An increase in mid-term global and regional RV strain beyond pre-operative values suggests positive RV remodeling and adaptation occurs in children post-PVR. Patients with larger pre-operative RV volumes had lower RV strain post-operatively. The impact of LV rotation on RV mechanics highlights the presence of ventriculo-ventricular interactions. These findings have important clinical implications in pediatric rTOF patients towards identifying pre-operative factors that predict RV post-operative remodeling.

Patients with prior vertebral or hip fractures treated with teriparatide in the Direct Assessment of Nonvertebral Fractures in Community Experience (DANCE) observational study.

SUMMARY: In patients in the Direct Assessment of Nonvertebral Fractures in Community Experience (DANCE) observational study with and without a prior vertebral or hip fracture, the incidence of nonvertebral fractures was lower with >6 months of teriparatide treatment than during the first 6 months. INTRODUCTION: Clinical evidence on the effect of teriparatide in patients with prior fracture is limited. In the DANCE observational study, the incidence of nonvertebral fragility fractures (NVFX) decreased significantly in patients receiving teriparatide for >6 months (6-24 months) versus >0 to ≤6 months (reference period). METHODS: We performed a post hoc analysis to assess the effect of teriparatide 20 µg/day in patients who entered DANCE with prior vertebral or hip fractures. The incidence of patients experiencing a NVFX for four 6-month intervals during and after treatment was compared with the reference period. RESULTS: Overall, 4085 patients received ≥1 dose of teriparatide. Of 3720 with sufficient data for efficacy analysis, 692 had prior vertebral fracture, including 179 with previous kyphoplasty/vertebroplasty; 290 had prior hip fracture. These patients were older, and those with prior vertebral fractures had more comorbid conditions at baseline than those without prior vertebral fractures. The incidence of patients experiencing NVFX declined over time in all patient groups. The fracture incidence rate declined 49 and 46%, respectively, in patients with and without prior vertebral fracture and was 63 and 46% lower in patients with previous kyphoplasty/vertebroplasty and without prior vertebral fracture. NVFX declined 43 and 48% in patients with and without prior hip fracture. The reduced incidence over time was consistent in the subgroups (all interaction p values >0.05). Patients with prior fracture were more likely to experience serious adverse events. CONCLUSION: The incidence of NVFX decreased over time in patients receiving teriparatide in DANCE regardless of prior fracture status.

Casein kinase 1 regulates Sprouty2 in FGF-ERK signaling.

Sprouty2 (SPRY2) is a potent negative regulator of receptor tyrosine kinase signaling, and is implicated as a tumor suppressor. SPRY2 inhibits FGF-RAS-ERK signaling by binding to growth factor receptor bound protein 2 (GRB2) during fibroblast growth factor receptor (FGFR) activation, disrupting the GRB2-SOS (son of sevenless) complex that transduces signals from FGFR to RAS. SPRY2 binding to GRB2 is modulated by phosphorylation but the key regulatory kinase(s) are not known. Prior studies identified the frequent presence of CK1 phosphorylation motifs on SPRY2. We therefore tested if CK1 has a role in SPRY2 phosphorylation and function. Loss of CK1 binding and inhibition of CK1 activity by two structurally distinct small molecules abrogated SPRY2 inhibition of FGF-ERK signaling, leading to decreased SPRY2 interaction with GRB2. Moreover, CK1 activity and binding are necessary for SPRY2 inhibition of FGF-stimulated neurite outgrowth in PC12 cells. Consistent with its proposed role as an inhibitor of FGF signaling, we find that CSNK1E transcript abundance negatively correlates with FGF1/FGF7 message in human gastric cancer samples. Modulation of CK1 activity may be therapeutically useful in the treatment of FGF/SPRY2-related diseases.

Randomised phase II trial of docetaxel and sunitinib in patients with metastatic gastric cancer who were previously treated with fluoropyrimidine and platinum.

BACKGROUND: Docetaxel is widely used as a chemotherapeutic agent for gastric cancer treatment. A combined regimen with sunitinib demonstrated a synergistic antitumour effect in a preclinical model. The aim of this study was to evaluate the efficacy and safety of this combination in patients with unresectable or metastatic advanced gastric cancer following failure of treatment with a fluoropyrimidine and platinum combination. METHODS: This open-label, phase II, randomised trial enrolled patients with unresectable or metastatic gastric cancer. Patients were assigned to either a docetaxel monotherapy arm (D only arm: 60 mg m(-2), every 3 weeks) or a combination arm (DS arm: docetaxel+sunitinib 37.5 mg every day). The primary end point of the study was time to progression and the secondary end points were overall response rate, disease control rate, overall survival, and toxicity profile. A pharmacokinetic study was also performed. RESULTS: A total of 107 patients were entered into the study. The TTP was not significantly prolonged in the DS arm when compared with the D only arm (DS vs D only arm: 3.9 months (95% confidence interval (CI) 2.9-4.9) vs 2.6 months (95% CI 1.8-3.5) (P=0.206). The hazard ratio for TTP was 0.77 (95% CI 0.52-1.16). However, the objective response rate was significantly higher in the DS arm (41.1% vs 14.3%, P=0.002). Patients in the DS arm experienced stomatitis, diarrhoea, and hand-foot syndrome more frequently. CONCLUSION: The addition of sunitinib to docetaxel did not significantly prolong TTP, although it significantly increased response.

SU-E-J-172: Development of a Video Guided Real-Time Patient Motion Monitoring System for Helical Tomotherpay.

PURPOSE: We developed a video image-guided real-time patient motion monitoring system for helical Tomotherapy (VGRPM-Tomo), and its clinical utility was evaluated using a motion phantom. METHODS: The VGRPM-Tomo consisted of three components: an image acquisition device consisting of two PC-cams, a main control computer with a radiation signal controller and warning system, and patient motion analysis software, which was developed in house. The system was designed for synchronization with a beam on/off trigger signal to limit operation during treatment time only and to enable system automation. In order to detect the patient motion while the couch is moving into the gantry, a reference image, which continuously updated its background by exponential weighting filter (EWF), is compared with subsequent live images using the real-time frame difference-based analysis software. When the error range exceeds the set criteria (δ_movement) due to patient movement, a warning message is generated in the form of light and sound. The described procedure repeats automatically for each patient. A motion phantom, which operates by moving a distance of 0.1, 0.2, 0.5, and 1.0 cm for 1 and 2 sec, respectively, was used to evaluate the system performance at maximum couch speed (0.196 cm/sec) in a Helical Tomotherapy (HD, Hi-art, Tomotherapy, USA). We measured the optimal EWF factor (a) and δ_movement, which is the minimum distance that can be detected with this system, and the response time of the whole system. RESULTS: The optimal a for clinical use ranged from 0.85 to 0.9. The system was able to detect phantom motion as small as 0.2 cm with tight δ_movement, 0.1% total number of pixels in the reference image. The measured response time of the whole system was 0.1 sec. CONCLUSIONS: The VGRPM-tomo can contribute to reduction of treatment error caused by the motion of patients and increase the accuracy of treatment dose delivery in HD. This work was supported by the Technology Innovation Program, 10040362, Development of an integrated management solution for radiation therapy funded by the Ministry of Knowledge Economy (MKE, Korea). This idea is protected by a Korean patent (patent no. 10-1007367).

SU-E-T-292: New Technique for Developing Proton Range Compensator Using Three-Dimensional Printer.

PURPOSE: A new system for manufacturing proton range compensator (PRC) was developed by using a three-dimensional printer (3DP). The physical accuracy and dosimetrical characteristics of the new PRC (PRC-3DP) was compared with conventional PRC (PRC-CMM) manufactured by computerized milling machine (CMM). METHODS: A PRC for brain cancer treatment, with passive scattered proton beam, was calculated in the TPS (Eclipse, Varian, USA) and its data was converted into a new format for 3DP (Projet HD3000, 3D Systems, USA), using the in-house developed software. PRC-3DP was printed with UV curable acrylic plastic, while PRC- CMM was milled into PMMA using a CMM (V-CNC500, CINCINNATI, USA). We measured the 5 randomly selected points for its physical thickness of both PRCs to evaluate its physical accuracy. Stopping power ratio (SPR), spread-out bragg peak (SOBP, 90∼90%) and distal fall-off (DFO, 20∼80%) at the central axis, +2.5, and 2.5 cm in the lateral direction, and FWHM of dose profile in depth 6, 8, and 10 cm were measured to evaluate for its dosimetrical characteristics. All measured data was compared with TPS data. RESULTS: There was no significant difference in the physical depths between the calculated and the measured value of both RPC-3DP and RPC-CMM (p<0.05). SPR of both PRC showed similarity in value (1.022) when compared with that of the water. Average difference of SOBP between the TPS and the measured data from both PRC was 0.3773±0.0075 and 0.2762±0.0235 cm, while DFO was 0.06±0.005 and 0.0471±0.0042 cm, respectively. Average differences of FWHM between the TPS and the measured data from PRC-3DP and PRC-CMM were 0.1799±0.025 and 0.137±0.0181 cm, respectively. There was no significant difference in dosimetrical characteristic between the RTP and both PRCs (p<0.05). CONCLUSIONS: Physical accuracy and dosimetrical characteristics of the PRC-3DP were comparable to that of the conventional PRC-CMM, while significant system minimization was provided. This work was supported by the Technology Innovation Program, 10040362, Development of an integrated management solution for radiation therapy funded by the Ministry of Knowledge Economy (MKE, Korea). This idea was applied for a Korea patent (no. 10-2012-0010812).

Effects of low-level arsenic exposure on urinary N-acetyl-ß-D-glucosaminidase activity.

This study was aimed to evaluate whether renal tubular function is impaired by exposure to relatively low concentrations of arsenic. Mean urinary arsenic concentrations and N-acetyl-ß-D-glucosaminidase (NAG) activities were compared among 365 and 502 Korean men and women, respectively, in relation to gender, smoking, alcohol consumption, and recent seafood consumption. The study subjects were divided into 4 groups according to urinary NAG activity and seafood consumption prior to urine sampling, and the correlation between arsenic concentration and urinary NAG activity was tested for each group. The mean urinary arsenic level was higher in women, non-smokers, and non-drinkers in comparison to men, smokers, and drinkers, respectively. Individuals who consumed seafood within 3 days prior to urine sampling showed a higher mean urinary arsenic level than those who did not. The correlation between urinary arsenic concentration and NAG activity in urine was significant only in subjects who did not consume seafood within 3 days prior to urine sampling and whose urinary NAG activity was 7.44 U/g creatinine (75th percentile) or higher. The urinary arsenic concentration was a significant determinant of urinary NAG activity in subjects with NAG activity higher than 7.44 U/g creatinine and especially in those who had not consumed seafood recently. These facts suggest that a relatively low-level exposure to inorganic arsenic produces renal tubular damage in humans.

Production of solid lipid nanoparticle suspensions using supercritical fluid extraction of emulsions (SFEE) for pulmonary delivery using the AERx system.

The aims of the current work included: development of a new production method for nanoparticles of water-insoluble drugs in combination with lipids, characterization of the nanoparticles and development of lipid nanosuspension formulations, and investigation of the feasibility of delivering the nanosuspensions as aerosols for inhalation using Aradigm's AERx Single Dose Platform (SDP) with micron-sized nozzles and the all mechanical AERx Essence with sub-micron-sized nozzles. The continuous SFEE method was used for particle precipitation of solid lipid nanoparticles (SLN). The method allowed for production of stable particulate aqueous suspensions of a narrow size distribution, with a volume mean diameter below 30 nm (D99% cumulative volume below 100 nm). Thus the particle size obtained was significantly smaller than previously has been achieved by other techniques. The residual solvent content in the final suspension was consistently below 20 ppm. Drug loading values between 10-20% w/w drug were obtained for model compounds ketoprofen and indomethacin in formulation with lipids such as tripalmitin, tristearin and Gelucire 50/13. It was observed that the loading capacity achieved was higher than the thermodynamic limit of the solubility of the drugs in molten lipids. Lipid nanosuspension formulations were successfully aerosolized using both of the AERx systems. As measured by both cascade impactor and laser diffraction, the aerosol fine particle fraction (FPF) was comparable to drug solution formulations typically used in these devices; i.e., greater than 90% of the aerosol mass resided in particles less than 3.5 mum aerodynamic diameter.

The effect of excess protamine on thrombelastography in vitro.

The aim of the current study was to assess the direct effect of protamine on conventional thrombelastography in vitro. Protamine was added to blood samples collected from 25 adult cardiac surgical patients prior to the induction of anaesthesia and after separation from cardiopulmonary bypass. The final protamine concentrations were 0 (control), 0.05 mg/ml, 0.1 mg/ml and 0.2 mg/ml (i.e. sufficient to reverse heparin 0, 5, 10 and 20 IU/ml respectively, assuming a 1:1 reversal ratio). In the pre-induction samples, protamine was associated with increases in r time and reductions in maximum amplitude (P<0.01). After bypass, the control samples demonstrated a heparin effect as expected, which was corrected by the addition of protamine 0.05 mg/ml. However, the higher concentrations of protamine were again associated with increases in r time and reductions in maximum amplitude (P<0.01). The results indicate that protamine has a direct anticoagulant effect on conventional thrombelastography in vitro. This effect occurs whether protamine is present alone, or whether protamine is present in excess after neutralization of heparin. Unless this effect is taken into account, excess protamine may confound the interpretation of conventional thrombelastography in cardiac surgical patients.

A novel mutation in the mitochondrial tRNA(Ser(AGY)) gene associated with mitochondrial myopathy, encephalopathy, and complex I deficiency.

PURPOSE: To identify molecular defects in a girl with clinical features of MELAS (mitochondrial encephalomyopathy and lactic acidosis) and MERRF (ragged-red fibres) syndromes. METHODS: The enzyme complex activities of the mitochondrial respiratory chain were assayed. Temporal temperature gradient gel electrophoresis was used to scan the entire mitochondrial genome for unknown mitochondrial DNA (mtDNA) alterations, which were then identified by direct DNA sequencing. RESULTS: A novel heteroplasmic mtDNA mutation, G12207A, in the tRNA(Ser(AGY)) gene was identified in the patient who had a history of developmental delay, feeding difficulty, lesions within her basal ganglia, cerebral atrophy, proximal muscle weakness, increased blood lactate, liver dysfunction, and fatty infiltration of her muscle. Muscle biopsy revealed ragged red fibres and pleomorphic mitochondria. Study of skeletal muscle mitochondria revealed complex I deficiency associated with mitochondrial proliferation. Real time quantitative PCR analysis showed elevated mtDNA content, 2.5 times higher than normal. The tRNA(Ser(AGY)) mutation was found in heteroplasmic state (92%) in the patient's skeletal muscle. It was not present in her unaffected mother's blood or in 200 healthy controls. This mutation occurs at the first nucleotide of the 5' end of tRNA, which is involved in the formation of the stem region of the amino acid acceptor arm. Mutation at this position may affect processing of the precursor RNA, the stability and amino acid charging efficiency of the tRNA, and overall efficiency of protein translation. CONCLUSION: This case underscores the importance of comprehensive mutational analysis of the entire mitochondrial genome when a mtDNA defect is strongly suggested.

Population pharmacokinetics of intravenous valproic acid in Korean patients.

OBJECTIVE: To determine population-based pharmacokinetic parameters for intravenous valproic acid, and the factors influencing these parameters, in Korean adults. METHODS: Valproic acid concentrations were obtained using a peak and trough sampling scheme for 102 Korean epileptic patients who were not taking concurrent antiepileptic medication. Three hundred and fifty-four serum concentrations were analysed according to a one-compartment model with a mixed effect modelling method (NONMEM Ver 5.0). The influence of body-weight (kg), height, daily valproic acid dose (mg/day), body mass index (kg/m2), sex, and age on volume of distribution (Vd) and clearance (CL) was assessed in the course of analysis. RESULTS: Vd and CL of valproic acid increased with body-weight. No significant influence of the other screened covariates was observed. The final regression model was: [equation: see text]. Interindividual variabilities (coefficient of variation) for CL and Vd were 32 and 18%, respectively. Residual error including intraindividual variability was 26.7%. CONCLUSION: The current results may be used as a basic reference to optimize drug therapy with intravenous valproic acid. Further research on the paediatric population is necessary to confirm the non-linearity of the relation between body-weight and Vd.

Relationship between the Val158Met polymorphism of catechol O-methyl transferase and breast cancer.

A case-control study was performed to assess the potential influence of catechol O-methyl transferase (COMT) genotype on the risk of breast cancer in Korean women. One hundred and sixty-three histologically confirmed incident breast cancer cases and 163 age- and menopausal status-matched control individuals with no present or previous history of cancer were selected as study subjects. COMT genetic polymorphism was determined by gel electrophoresis after NlaIII enzyme digestion of amplified DNA. Odds ratios and 95% confidence intervals were estimated by unconditional logistic regression after adjustment for known or suspected risk factors of breast cancer. Women with at least one COMT lower enzyme activity associated allele (COMT-L) were at elevated risk for breast cancer (OR = 1.7, 95% CI = 1.04-2.78) compared with those homozygous for high enzyme activity associated COMT-H alleles. Among women with low (> or = 23.1) body mass index the COMT-L allele containing genotypes posed a marginally significant increased risk of breast cancer compared to the COMT-HH genotype (OR = 1.8, 95% CI = 0.95-3.48). Women with at least one COMT-L allele who had experienced a full-term pregnancy when aged over 30 years or were nulliparous had 2.7-fold increased risk; however, this increase did not reach statistical significance (OR = 2.7, 95% CI = 0.64-11.35). Furthermore, never-drinking and never-smoking women with at least one COMT-L allele were at increased risk of breast cancer compared to those with COMT-HH genotype with ORs of 2.0 (95% CI = 1.23-3.38) and 1.7 (95% CI = 1.04-2.62), respectively. These results are consistent with studies showing that COMT genotype of lower enzyme activity might be related to increase in risk of breast cancer, and extend this finding to Korean women.

Molecular cloning and characterization of pig immunoreceptor DAP10 and NKG2D.

Pig immunoreceptor DAP10 cDNA was cloned from a peripheral blood lymphocyte (PBL) cDNA library using human DAP10 cDNA as a probe. The length of the pig DAP10 cDNA is 465 bp and it contains an open reading frame of 237 bp. The predicted polypeptide sequence is 79 amino acids, consisting of an 18-amino acid leader, a 16-amino acid extracellular domain, a 24-amino acid transmembrane segment, and a 21-amino acid cytoplasmic domain. The amino acid sequence of pig DAP10 has 68% and 78% sequence identity with human DAP10 and mouse DAP10, respectively. Pig DAP10 has a conserved aspartic acid in the transmembrane domain, two cysteines in the extracellular domain, and a phophatidylinositol-3 kinase-binding site (YxxM) in the cytoplasmic region. Genomic organization reveals that pig DAP10 comprises four exons and three introns. Pig DAP10 and DAP12 are genetically linked on Chromosome (Chr) 6 at 6q21 in opposite transcriptional orientation, separated by 152 bp. In Northern blot analysis, DAP10 transcripts were detected predominantly in lymphohematopoietic tissues. Pig NKG2D cDNA has an open reading frame of 642 bp. Its expected polypeptide sequence is 214 amino acids. Pig NKG2D has 66% sequence identity with human NKG2D and 56% identity with mouse NKG2D. The NKG2D gene maps to pig Chr 5q25. RT-PCR analysis reveals that pig NKG2D transcripts are expressed in PBLs, NK cells, macrophages, and monocytes. When transiently transfected into COS-7 cells, pig NKG2D requires DAP10 for cell surface expression.

Molecular cloning and expression pattern of porcine myeloid DAP12-associating lectin-1.

DAP12 is an ITAM-bearing membrane protein that is associated with activating receptors in natural killer cells, granulocytes, macrophages, and monocytes. Myeloid DAP12-associating lectin-1 (MDL-1) is a type II membrane protein that associates with DAP12. In this study, we report the molecular cloning of two isoforms of porcine MDL-1 cDNA from pulmonary alveolar macrophages. The porcine MDL-1 short form has 165 amino acids and 70% sequence identity with the mouse MDL-1 short form. The long form has 20 more amino acids in the stalk region and 71% sequence identity with human MDL-1 and 67% with the mouse MDL-1 long form. Porcine MDL-1 contains a conserved lysine in the transmembrane domain. There are six putative N-linked glycosylation sites in the MDL-1 long form. MDL-1 transcripts were detected exclusively in macrophages and monocytes by RT-PCR. When transfected into 293 cells, porcine MDL-1 is expressed on the cell surface associated with DAP12.

Assay of omeprazole and omeprazole sulfone by semi-microcolumn liquid chromatography with mixed-function precolumn.

A column-switching system based on semi-microcolumns was used for direct analysis of omeprazole and omeprazole sulfone in human plasma samples. Plasma samples were injected into a mixed-function (MF Ph-1) column (35 mmx4.6 mm I.D.) to remove proteins and other non-specific peak producing substances from the analyte-containing time zone. The analyte-containing fraction was thereafter transferred to a C-18 semi-microcolumn (250 mmx1.5 mm I.D.) after concentration at the C-18 intermediate column. The absorbance at 302 nm in a ultraviolet (UV) detector was recorded to measure the concentration. The detection limit for omeprazole and omeprazole sulfone in the present method was 10 ng/ml. Interbatch variation (coefficient of variation) of the QC samples spanned less than 10% and intra-batch variation less than 2%. The recovery ratios of omeprazole and omeprazole sulfone were over 98%. The current method can be used as a simpler procedure with similar sensitivity and reproducibility as previously reported methods.

Effect of omeprazole on the pharmacokinetics of moclobemide according to the genetic polymorphism of CYP2C19.

BACKGROUND: Moclobemide, an antidepressant with selective monoamine oxidase-A inhibitory action, is known to be metabolized by CYP2C19 and is also reported to be an inhibitor of CYP2C19, CYP2D6, and CYP1A2. To confirm the involvement of CYP2C19, we performed a pharmacokinetic interaction study. METHODS: The effect of omeprazole on the pharmacokinetics of moclobemide was studied in 16 healthy volunteers. The volunteer group comprised 8 extensive metabolizers and 8 poor metabolizers of CYP2C19, which was confirmed by genotyping. Subjects were randomly allocated into two sequence groups, and a single-blind, placebo-controlled, two-period crossover study was performed. In study I, a placebo was orally administered for 7 days. On the eighth morning, 300 mg of moclobemide and 40 mg of placebo were coadministered with 200 mL of water, and a pharmacokinetic study was performed. During study II, 40 mg of omeprazole was given each morning instead of placebo, and pharmacokinetic studies were performed on the first and eighth day with 300 mg of moclobemide coadministration. RESULTS: The inhibition of moclobemide metabolism was significant in extensive metabolizers even after a single dose of omeprazole. After daily administration of omeprazole for 1 week, the pharmacokinetic parameters of moclobemide and its metabolites in extensive metabolizers changed to values similar to those in poor metabolizers. In poor metabolizers, no remarkable changes in the pharmacokinetic parameters were observed. CONCLUSION: Our results show that CYP2C19 is an important enzyme in the elimination of moclobemide and that it is extensively inhibited by omeprazole in extensive metabolizers, but not in poor metabolizers.

Longitudinal assessment of Pseudomonas aeruginosa in young children with cystic fibrosis.

Pseudomonas aeruginosa lung infection is an important cause of morbidity and mortality in cystic fibrosis (CF). Longitudinal assessment of the phenotypic changes in P. aeruginosa isolated from young children with CF is lacking. This study investigated genotypic and phenotypic changes in P. aeruginosa from oropharynx (OP) and bronchoalveolar lavage fluid (BALF) in a cohort of 40 CF patients during the first 3 years of life; antibody response was also examined. A high degree of genotypic variability was identified, and each patient had unique genotypes. Early isolates had a phenotype distinct from those of usual CF isolates: generally nonmucoid and antibiotic susceptible. Genotype and phenotype correlated between OP and BALF isolates. As determined by culture, 72.5% of patients demonstrated P. aeruginosa during their first 3 years. On the basis of combined culture and serologic results, 97.5% of patients had evidence of infection by age 3 years, which suggests that P. aeruginosa infection occurs early in CF and may be intermittent or undetectable by culture.

Molecular cloning, gene structure, and expression pattern of pig immunoreceptor DAP12.

Natural killer (NK) cells express receptors for MHC class I that contain immunoreceptor tyrosine-based inhibitory motif (ITIM) sequences in their cytoplasmic domain. Whereas these receptors inhibit NK cell cytotoxicity, certain isoforms of these NK receptors (e.g., KIR2DS, CD94/NKG2C, and Ly49D) do not have ITIMs, but associate with DAP12 and activate NK cell function. We cloned pig DAP12 cDNA from a pig peripheral blood lymphocyte (PBL) cDNA library using human DAP12 cDNA as a probe. The length of the pig DAP12 cDNA is 526 bp and contains an open reading frame of 324 bp. It has 79% identity with the human DAP12 cDNA sequence in the coding region and 73% identity with mouse DAP12 cDNA. The predicted polypeptide sequence of pig DAP12 is 108 amino acids, being composed of a 23-amino acid leader, a 14-amino acid extracellular domain, a 24-amino acid transmembrane segment, and a 47-amino acid cytoplasmic region. The amino acid sequence of pig DAP12 has 74% and 71% sequence identity with human DAP12 and mouse DAP12, respectively. Pig DAP12 has a conserved aspartic acid in the transmembrane region, and two conserved cysteine residues in the extracellular domain. It also contains an immunoreceptor tyrosine-based activation motif sequence in the cytoplasmic region. Genomic organization reveals that pig DAP12 consists of five exons and four introns. Southern blot analysis of pig genomic DNA revealed that DAP12 is a single-copy gene. In Northern blot analysis, DAP12 transcripts were detected in spleen, liver, thymus, and lymph node. DAP12 transcripts are expressed not only in PBLs, but also in granulocytes, macrophages, and monocytes.