Protocol of BEYOND trial: Clinical BEnefit of sodium-glucose cotransporter-2 (SGLT-2) inhibitors in rhYthm cONtrol of atrial fibrillation in patients with diabetes mellitus.

SGLT-2 inhibitor, traditionally used for glycemic control, has several beneficial effects that can help manage heart failure (HF). SGLT-2 inhibitors reduce the risk of cardiovascular mortality in patients with HF. As atrial fibrillation (AF) is closely associated with HF and diabetes mellitus (DM) is a risk factor for AF, we assume that SGLT-2 inhibitors will also show therapeutic benefits regarding AF, especially for rhythm control. This trial has a multicenter, prospective, open, blinded endpoint design. It is a 1:1 randomized and controlled study. A total of 716 patients who are newly diagnosed of AF and DM within 1 year will be enrolled from 7 tertiary medical centers. The trial is designed to compare the effects of SGLT-2 inhibitors and other oral hypoglycemic agents on atrial rhythm control in patients with AF and DM. The primary outcome is the recurrence of AF within a year (including post-antiarrhythmic drugs (AAD) or ablation). The secondary outcomes are the ablation rate within a year, change in AF burden, size of the left atrium, NT-proBNP, the AF symptom score, and the quality of life. This trial will prospectively evaluate the effect and safety of SGLT-2 inhibitors on AF rhythm control in patients with DM. It will provide an invaluable dataset on rhythm control in AF with DM for future studies and offer novel information to assist in clinical decisions. (BEYOND trial, ClinicalTrials.gov number: NCT05029115. https://clinicaltrials.gov/ct2/show/NCT05029115).

Very Elderly Patients With Acute Coronary Syndromes Treated With Percutaneous Coronary Intervention.

BACKGROUND: The rates of very elderly patients (≥85 years old) undergoing percutaneous coronary intervention (PCI) for acute coronary syndromes (ACS) are rapidly increasing. They are under-represented in clinical trials, and those who are included may not reflect the real-world population. We aim to review the clinical characteristics of very elderly patients undergoing PCI for ACS and identify factors associated with adverse outcomes. METHOD: All very elderly patients undergoing PCI for ACS in the Auckland region between January 2014 to December 2016 were included. Baseline clinical and procedural details were obtained, and the primary endpoint was all-cause mortality measured up to a maximum of 4 years. Secondary endpoints include recurrent myocardial infarction, unplanned revascularisation, stroke and major bleeding. RESULTS: A total of 186 patients were included for analysis (mean age 87.6±2.8 years, 51.6% male). Indications for PCI were ST-elevation myocardial infarction (STEMI) in 74 (39.8%), non-ST elevation myocardial infarction (NSTEMI) in 97 (52.2%) and unstable angina in 15 patients (8.1%). Successful PCI was completed in 180 patients. At a maximal follow-up of 4 years (mean 23.4 mo), the rates of all-cause mortality and recurrent myocardial infarction were 22.0% and 14.0% respectively. The risk of mortality was increased by the presence of diabetes (44.8% vs 17.8%, HR=3.0, 95%CI: 1.6-5.9, p=0.001), STEMI (33.8% vs 13.5%, HR=3.1, 95%CI: 1.6-5.9, p=0.001), and reduced eGFR (every -10 mL/min/1.73m2, HR=1.7, 95%CI: 1.3-2.1, p<0.0001). Major bleeding events while on dual antiplatelet therapy as defined by Bleeding Academic Research Consortium score ≥3 occurred in 14 patients (7.5%; 8 on ticagrelor, 6 on clopidogrel). CONCLUSION: Very elderly patients who undergo PCI for ACS have acceptable survival outcomes. STEMI, diabetes and impaired renal function were predictive of mortality in this elderly cohort.

Intention to use light-rail transit in Houston, Texas, United States: Findings from the Travel-Related Activity in Neighborhoods study.

Using data from the Houston Travel-Related Activity in Neighborhoods (TRAIN) study, this study examined how various factors affect whether individuals intend to use newly opened light-rail transit (LRT) lines in Houston. The Houston TRAIN study is a natural experiment on the effect of new LRT lines on both transit use and physical activity. A mixed binary logit model was developed based on a dichotomous dependent variable and rich set of variables, including sociodemographic factors, health status, travel behavior and technology, and attitudes and perceptions. The mixed model also allowed accounting for the unobserved heterogeneity across individuals in their sensitivity to observed variables. The results indicated the important role of various factors influencing the decision on intent to use the new LRT lines. In general, demographics mattered but to a lower extent than psychological or personality-related variables. For example, attitudes and perceptions toward the public transit system and consciousness of physical activities derived by using public transit were important factors. Personal health constraints negatively influenced intention to use, while experience with the public transport system was among the positive indicators. The findings show the potential of future interventions in this community to promote use of the new system, such as educational campaigns that improve perceptions of public transit use and clarify the benefits of being active. While providing growing evidence that cognitive variables are important in measuring intention to use public transit, the results emphasize the positive role of efforts integrating transportation and health to develop effective and sustainable solutions.

A recombinant human immunoglobulin with coherent avidity to hepatitis B virus surface antigens of various viral genotypes and clinical mutants.

The hepatitis B virus (HBV) envelope is composed of a lipid bilayer and three glycoproteins, referred to as the large (L), middle (M), and small (S) hepatitis B virus surface antigens (HBsAg). S protein constitutes the major portion of the viral envelope and an even greater proportion of subviral particles (SVP) that circulate in the blood. Recombinant S proteins are currently used as a preventive vaccine, while plasma fractions isolated from vaccinated people, referred to as hepatitis B immune globulin (HBIG), are used for short-term prophylaxis. Here, we characterized a recombinant human IgG1 type anti-S antibody named Lenvervimab regarding its binding property to a variety of cloned S antigens. Immunochemical data showed an overall consistent avidity of the antibody to S antigens of most viral genotypes distributed worldwide. Further, antibody binding was not affected by the mutations in the antigenic 'a' determinant found in many clinical variants, including the immune escape mutant G145R. In addition, mutations in the S gene sequence that confer drug resistance to the viral polymerase did not interfere with the antibody binding. These results support for a preventive use of the antibody against HBV infection.

Efficient Atomic-Resolution Uncertainty Estimation for Neural Network Potentials Using a Replica Ensemble.

Neural network potentials (NNPs) are gaining much attention as they enable fast molecular dynamics (MD) simulations for a wide range of systems while maintaining the accuracy of density functional theory calculations. Since NNP is constructed by machine learning on training data, its prediction uncertainty increases drastically as atomic environments deviate from training points. Therefore, it is essential to monitor the uncertainty level during MD simulations to judge the soundness of the results. In this work, we propose an uncertainty estimator based on the replica ensemble in which NNPs are trained over atomic energies of a reference NNP that drives MD simulations. The replica ensemble is trained quickly, and its standard deviation provides atomic-resolution uncertainties. We apply this method to a highly reactive silicidation process of Si(001) overlaid with Ni thin films and confirm that the replica ensemble can spatially and temporally trace simulation errors at atomic resolution, which in turn guides the augmentation of the training set. The refined NNP completes a 3.6 ns simulation without any noticeable problems. By suggesting an efficient and atomic-resolution uncertainty indicator, this work will contribute to achieving reliable MD simulations by NNPs.

Understanding Potential Exposure of Bicyclists on Roadways to Traffic-Related Air Pollution: Findings from El Paso, Texas, Using Strava Metro Data.

As bicycling on roadways can cause adverse health effects, there is an urgent need to understand how bicycle routes expose bicyclists to traffic emissions. Limited resources for monitoring reveal that bicycle travel patterns may constrain such understanding at the network level. This study examined the potential exposure of bicyclists to traffic-related air pollution in El Paso, Texas, using Strava Metro data that revealed bicycle patterns across the city networks. An initial spatial mapping analysis was conducted to explore the spatial patterns of bicycling and traffic pollutant emission, followed by exploratory descriptive statistics. A spatial bicycle model was then developed to explore factors influencing bicycling activity in El Paso. Analysis results indicated significant associations between greater bicycle volume and both higher levels of particulate matter (PM2.5) emissions and more frequent bus services, implying adverse health concerns related to traffic-related air pollution. The results also indicated significant effects of various environmental characteristics (e.g., roadway, bicycle infrastructure, topography, and demographics) on bicycling. The findings encourage extending this study to provide guidance to bicyclists whose regular trips take place on heavily trafficked roads and during rush hours in this region and to evaluate the net health impacts of on-road bicycling for the general population.

Identification of ground-state spin ordering in antiferromagnetic transition metal oxides using the Ising model and a genetic algorithm.

We identify ground-state collinear spin ordering in various antiferromagnetic transition metal oxides by constructing the Ising model from first-principles results and applying a genetic algorithm to find its minimum energy state. The present method can correctly reproduce the ground state of well-known antiferromagnetic oxides such as NiO, Fe2O3, Cr2O3 and MnO2. Furthermore, we identify the ground-state spin ordering in more complicated materials such as Mn3O4 and CoCr2O4.

Quantification of Etodolac in Human Plasma for Pharmacokinetics and Bioequivalence Studies in 27 Korean Subjects.

OBJECTIVE: We developed a simple and validated liquid chromatography tandem mass spectrometry( LC-MS/MS) for quantification of etodolac using pioglitazone as an internal standard (IS) to assess pharmacokinetics and to appraise bioequivalence of two formulations of etodolac (reference and tested) in 27 healthy Korean subjects. METHODS: Isocratic mobile phase consisted of 10 mM ammonium formate and acetonitrile were used to separate the analytes on a Gemini C18 column. Also, analytes were analyzed by MS/MS in multiple reaction monitoring (MRM) mode using the transitions of (M+H)+ ions, m/z 288.2→ 172.3 and m/z 357.1→ 134.2 for quantification of etodolac and IS each. The standard calibration curves displayed significant linearity within the range of 0.2-30.0 µ g/mL (r2=0.9956, 1/x2 weighting) with LLOQ of 0.1 µg/mL. RESULTS: The retention times of etodolac and the IS were 0.77 min and 0.57 min each, indicating the high-throughput potential of the proposed method. The pharmacokinetic parameters were calculated from the plasma samples and data form the reference and test drugs were represented as follows; Area under plasma concentration-time curve (AUCt) (78.03 vs. 84.00 µgxh/mL), AUC∞ (86.67 vs. 93.92 µgxh/mL), maximal plasma concentration (Cmax) (19.49 vs. 18.94 µg/mL), time for maximal concentrations (Tmax) (2.13 vs. 2.26 h), Plasma elimination half-life (T1/2) (8.12 vs. 8.47 h), elimination rate constant (λz) (0.0853 vs. 0.0818 h-1). Pharmacokinetic parameters with 90% confidence interval fall within the bioequivalence range of 80-125%. CONCLUSION: Thus, the new testified method was successfully applied for the pharmacokinetic and bioequivalence studies for two etodolac formulations.

Use of a Target-Mediated Drug Disposition Model to Predict the Human Pharmacokinetics and Target Occupancy of GC1118, an Anti-epidermal Growth Factor Receptor Antibody.

GC1118 is an anti-epidermal growth factor receptor (EGFR) monoclonal antibody that is currently under clinical development. In this study, the pharmacokinetics (PK) of GC1118 were modelled in monkeys to predict human PK and receptor occupancy (RO) profiles. The serum concentrations of GC1118 and its comparator (cetuximab) were assessed in monkeys with a non-compartmental analysis and a target-mediated drug disposition (TMDD) model after intravenous infusion (3-25 mg/kg) of these drugs. The scaling exponent of the EGFR synthesis rate was determined using a sensitivity analysis. The human cetuximab exposures were simulated by applying different exponents (0.7-1.0) for the EGFR synthesis rate in the allometric monkey PK model. Simulated Cmax and area under the curve values therein were compared with those previously reported in the literature to find the best exponent for the EGFR synthesis rate in human beings. The TMDD model appropriately described the monkey PK profile, which showed a decrease in clearance (CL; 1.2-0.4 ml/hr/kg) as the dose increased. The exponents for CL (0.75) and volume of distribution (Vd; 1.0) were used for the allometric scaling to predict human PK. The allometric coefficient for the EGFR synthesis rate chosen by the sensitivity analysis was 0.85, and the RO profiles that could not be measured experimentally were estimated based on the predicted concentrations of the total target and the drug-target complex. Our monkey TMDD model successfully predicts human PK and RO profiles of GC1118 and can be used to determine the appropriate dose for a first-in-human study investigating this drug.

GC1118, an Anti-EGFR Antibody with a Distinct Binding Epitope and Superior Inhibitory Activity against High-Affinity EGFR Ligands.

The EGFR-targeted monoclonal antibodies are a valid therapeutic strategy for patients with metastatic colorectal cancer (mCRC). However, only a small subset of mCRC patients has therapeutic benefits and there are high demands for EGFR therapeutics with a broader patient pool and more potent efficacy. In this study, we report GC1118 exhibiting a different character in terms of binding epitope, affinity, mode of action, and efficacy from other anti-EGFR antibodies. Structural analysis of the EGFR-GC1118 crystal complex revealed that GC1118 recognizes linear, discrete N-terminal epitopes of domain III of EGFR, critical for EGF binding but not overlapping with those of other EGFR-targeted antibodies. GC1118 exhibited superior inhibitory activity against high-affinity EGFR ligands in terms of EGFR binding, triggering EGFR signaling, and proliferation compared with cetuximab and panitumumab. EGFR signaling driven by low-affinity ligands, on the contrary, was well inhibited by all the antibodies tested. GC1118 demonstrated robust antitumor activity in tumor xenografts with elevated expression of high-affinity ligands in vivo, whereas cetuximab did not. Considering the significant role of high-affinity EGFR ligands in modulating tumor microenvironment and inducing resistance to various cancer therapeutics, our study suggests a potential therapeutic advantage of GC1118 in terms of efficacy and a range of benefited patient pool. Mol Cancer Ther; 15(2); 251-63. ©2015 AACR.

Drug synergism of antihypertensive action in combination of telmisartan with lercanidipine in spontaneous hypertensive rats.

To examine drug synergism between angiotensin II AT1-receptor blocker and Ca(2+) channel blocker for lowering blood pressure (BP), telmisartan and lercanidipine were orally injected into to telemetered-spontaneous hypertensive rats and BP was monitored. The highest doses of both drugs (7.66 mg/kg of telmisartan and 1.92 mg/kg of lercanidipine) were clinically relevant at 80 and 20 mg human equivalent doses, respectively, and denoted as dose 1. After constructing the dose-response curve using 0 (vehicle-treated control), 1/16, 1/8, 1/4, 1/2 and 1 doses, all possible combinations of both drugs were tested. Drug synergism in combination therapy of telmisartan with lercanidipine was assessed by calculating the interaction index (γ) as evaluated by γ < 1. We found statistically significant drug synergism in the investigated (telmisartan: lercandipine) combinations of (1/8:1/4), (1/4:1) and (1/8:1). Our results suggest that the combination therapy of telmisartan and lercanidipine at lower doses are effective in lowering BP, and also reduce side effects caused by maximal doses of each drug. Therefore, drug combination of AT1-receptor blocker with Ca(2+) channel blocker is a clinically important tool for the management of hypertension and hypertension-related cardiovascular risks.

Adenosine amine congener mitigates noise-induced cochlear injury.

Hearing loss from noise exposure is a leading occupational disease, with up to 5% of the population at risk world-wide. Here, we present a novel purine-based pharmacological intervention that can ameliorate noise-induced cochlear injury. Wistar rats were exposed to narrow-band noise (8-12 kHz, 110 dB SPL, 2-24 h) to induce cochlear damage and permanent hearing loss. The selective adenosine A(1) receptor agonist, adenosine amine congener (ADAC), was administered intraperitoneally (100 microg/kg/day) at time intervals after noise exposure. Hearing thresholds were assessed using auditory brainstem responses and the hair cell loss was evaluated by quantitative histology. Free radical damage in the organ of Corti was assessed using nitrotyrosine immunohistochemistry. The treatment with ADAC after noise exposure led to a significantly greater recovery of hearing thresholds compared with controls. These results were upheld by increased survival of sensory hair cells and reduced nitrotyrosine immunoreactivity in ADAC-treated cochlea. We propose that ADAC could be a valuable treatment for noise-induced cochlear injury in instances of both acute and extended noise exposures.

Effect of electrokinetic stabilizers on the physicochemical properties of propofol emulsions.

The aims of the present study were to elucidate the potential mechanism of propofol emulsion destabilization following the addition of lidocaine, and to evaluate the effects of various electrokinetic stabilizers on the physicochemical properties of lidocaine-propofol emulsions. The assessments included pH observations and determination of the maximum globule diameter (MGD) and zeta potential (ZP). The MGD of propofol emulsions increased up to several tens mum following the addition of 50 mg of lidocaine to 200mg of propofol, and the proposed destabilization mechanism involves localization of protonated lidocaine molecules between lecithin molecules in propofol emulsions, which consequently leads to increased ZP. The ZP of propofol emulsions containing acidic amino acid or neutral amino acid increased following the addition of lidocaine, and a charge reversal occurred. Therefore, the MGD of emulsions increased to several tens (m. However, the MGD of emulsions that contained basic amino acids or basic compounds remained below 5 (m, despite the addition of large amounts lidocaine (50 mg), and the ZP did not pass through the point of zero charge. In conclusion, our results provide not only further insight into the physical stability of propofol emulsions containing lidocaine, but also a better understanding of the administration of propofol in existing applications.

High performance liquid chromatographic analysis of rabeprazole in human plasma and its pharmacokinetic application.

A simple and sensitive HPLC method for the analysis of rabeprazole in plasma is described using UV detection in the presence of lorazepam as the internal standard. Rabeprazole and lorazepam were extracted with ethyl ether and quantitated using a reverse-phase C(18) column. The method was specific as there were no interfering peaks in the human plasma eluting at the retention times of rabeprazole and lorazepam. The method was fully validated in human plasma for the concentration range of 20.0-1000.0 ng/ml. The correlation coefficients were greater than 0.999. Extraction recoveries were 72.3% for the drug and 79.1% for the internal standard. The method was simple, reliable, and accurate for the quantitation of rabeprazole in human plasma. The same plasma samples, which were collected in healthy male volunteers administered a 20 mg tablet of Pariet, were analyzed by HPLC and LC/MS/MS. As a result of that, there was no significant difference between pharmacokinetic parameters. The suitability of HPLC method for pharmacokinetic studies was verified by determining the relevant pharmacokinetic parameters.

Application of instrumental evaluation of color for the pre-formulation and formulation of rabeprazole.

The aims of this study were to fast screen the compatibility of rabeprazole and excipients using a spectrocolorimeter and to examine the relationship between the color change value and drug contents/drug degradation products in solid dosage forms. The color change values of rabeprazole-excipient mixtures were measured using a spectrocolorimeter, with six tablet formulations compressed using a single-punch instrumental tablet press. The rabeprazole and degradation products contents in the tablets were analyzed using an HPLC method, with the color change values of the tablets measured using spectrocolorimetery for 4 weeks. These experiments indicated that the instrumental evaluation of color was a speedy, simple and useful tool in the determination of the interaction between the drug and excipients, as well as in the formulation of solid dosage forms. The relationships of the % reduced drug contents versus the color change value, and those of the % drug degradation products versus the color change value were exponentially increased in formulations containing zinc stearate. On stress testing, the color change value of rabeprazole was inconsistent with previous reports, as the degradation of rabeprazole can be greatly influenced by humidity as well as temperature. Consequently, these results highlight the potential of color formation in the application of pre-formulation and formulation of drugs.

Scalable production of adeno-associated virus type 2 vectors via suspension transfection.

Vectors derived from adeno-associated virus type 2 (AAV2) are promising gene delivery vehicles, but it is still challenging to get the large number of recombinant adeno-associated virus (rAAV) particles required for large animal and clinical studies. Current transfection technology requires adherent cultures of HEK 293 cells that can only be expanded by preparing multiple culture plates. A single large-scale suspension culture could replace these multiple culture preparations, but there is currently no effective co-transfection scheme for generating rAAV from cells in suspension culture. Here, we weaned HEK 293 cells to suspension culture using hydrogel-coated six-well culture plates and established an efficient transfection strategy suitable for these cells. Then the cultures were gradually scaled up. We used linear polyethylenimine (PEI) to mediate transfection and obtained high transfection efficiencies ranging from 54% to 99%, thereby allowing efficient generation of rAAV vectors. Up to 10(13) rAAV particles and, more importantly, up to 10(11) infectious particles were generated from a 2-L bioreactor culture. The suspension-transfection strategy of this study facilitates the homogeneous preparation of rAAV at a large scale, and holds further potential as the basis for establishing a manufacturing process in a larger bioreactor.

Adeno-associated virus-mediated expression of apolipoprotein (a) kringles suppresses hepatocellular carcinoma growth in mice.

Hepatocellular carcinoma (HCC) constitutes more than 90% of all primary liver cancers. HCC is a hypervascular tumor that develops from dedifferentiation of small avascular HCC and is therefore a good target for anti-angiogenic gene therapy. Recent studies have identified apolipoprotein(a) [apo(a)] kringles LK68 and LK8 (LKs) as having a potential antiangiogenic and anti-tumor activity, and the current study evaluates the therapeutic potential of gene therapy with recombinant adeno-associated virus carrying genes encoding LKs (rAAV-LK) in the treatment of hypervascular HCC. We generated rAAV-LK to obtain persistent transgene expression in vivo, which is essential for anti-angiogenic therapy. The rAAV-produced LKs substantially inhibited proliferation and migration of human umbilical vein endothelial cells (HUVECs) in vitro, validating their anti-angiogenic potential. Intramuscular administration of rAAV-LK gave 60% to 84% suppression (P < .05) of tumor growth in mice bearing subcutaneously transplanted HCC derived from Huh-7 and Hep3B cells, respectively. Histological and immunohistochemical analyses of HCC tumor sections showed that a single administration of rAAV-LK gave rise to persistent expression of LKs that inhibited tumor angiogenesis and triggered tumor apoptosis, and, thus, significantly suppressed tumor growth. The administration of rAAV-LK provided a significant survival benefit (P < .05), and 3 of 10 rAAV-LK-treated mice were still alive without visible tumors and without clinical symptoms 188 days after treatment. In conclusion, rAAV-LK is a potential candidate for anti-angiogenic gene therapy in the treatment of HCC.

The effect of lidocaine on the globule size distribution of propofol emulsions.

In this study, we sought to determine the globule size distribution of a propofol/lidocaine mixture as a function of lidocaine concentration and time elapsed after mixing in a standard formulation of propofol emulsion (Diprivan) and in a new formulation containing L-lysine to improve stability. The globule size was measured with a laser diffraction technique. The median diameter of the globule size in 20 mL of Diprivan immediately after the addition of 0, 10, 20, 30, 40, and 50 mg of lidocaine was similar to that of chylomicrons, ranging from 0.28 +/- 0.01 micro m to 0.30 +/- 0.02 micro m, over the whole range of lidocaine concentration. However, the maximum diameter increased slightly (from 0.97 +/- 0.01 micro m to 2.90 +/- 0.07 micro m) as the concentration of lidocaine increased. At 6 h after adding lidocaine, the maximum globule size had increased slightly (to 2.98 +/- 0.04 micro m) with 20 mg of lidocaine and increased considerably (to 51.76 +/- 0.62 micro m) when 30 mg of lidocaine was added. At 2 h after the addition of 50 mg of lidocaine, the maximum globule diameter had increased to 52.2 +/- 1.92 micro m. The maximum globule diameter in the propofol emulsion to which L-lysine was added as a stabilizer did not exceed 3.0 micro m even when the largest amount of lidocaine was added. This study demonstrated that when 30 mg of lidocaine was added to 20 mL of Diprivan and the solution was left for a period of time, the globule size increased. Its increase was minimized by the addition of L-lysine to the propofol emulsion.

Shuttle PCR-based cloning of the infectious adeno-associated virus type 5 genome.

Adeno-associated virus type 5 (AAV5), which is distinct from the other serotypes of AAV, has attracted considerable interest as a premier gene delivery vector. As do the other serotypes, AAV5 contains its 4.7 kb-sized, single-stranded genome flanked with inverted terminal repeats (ITRs) in a hairpin conformation, which serves frequently as pause and arrest sites for DNA polymerases during PCR. To amplify the full-length of the AAV5 genome in single step, we established a shuttled, long and accurate PCR (LA-PCR) procedure in the present study. Furthermore, helper oligonucleotides, which hybridize with the palindromic sequence elements in ITR, were designed and employed in PCR to prevent the formation of hairpin structures by highly GC-rich ITRs. Consequently, a 4.7 kb-sized PCR product was amplified successfully, and cloned into a pBluescript II KS(+) plasmid. Six plasmids, harboring the full-length AAV5 genome, rescued wild type AAV5 viruses on transfection to HeLa and HEK 293 cells, which were co-infected with helper adenoviruses. Western and Southern blot analyses supported further the fact that the pAAV5 plasmids harbored the full-length AAV5 genome. The PCR method described in this study is applicable for the cloning of genomes containing variable palindromic structures, in addition to AAV genomes of other serotypes.

Interspecies comparison of the oral absorption of itraconazole in laboratory animals.

The oral absorption and disposition of itraconazole were studied in rats, rabbits and dogs. Serum levels of itraconazole and its active metabolite, hydroxyitraconazole, were determined by a validated HPLC method. The absorption of itraconazole was relatively rapid in rats and dogs but was slower in rabbits. The terminal elimination half-life (T 1/2lambda(z)), time to the peak concentration (Tmax), dose and weight normalized area under the curve (AUC) and the peak concentration (Cmax) of itraconazole found in the dog were comparable to those reported in humans. As in humans, the metabolite to parent drug AUC ratios in rats and dogs were greater than unity but was less in rabbits. The dog appears to be an appropriate animal model while the rat, not the rabbit, may be used as an alternative animal model in predicting the oral absorption of itraconazole in humans.