Assessing and mitigating pH-mediated DDI risks in drug development - formulation approaches and clinical considerations.

pH-mediated drug-drug interactions (DDI) is a prevalent DDI in drug development, especially for weak base compounds with highly pH-dependent solubility. FDA has released a guidance on the evaluation of pH-mediated DDI assessments using in vitro testing and clinical studies. Currently, there is no common practice of ways of testing across the academia and industry. The development of biopredictive method and physiologically-based biopharmaceutics modeling (PBBM) approaches to assess acid-reducing agent (ARA)-DDI have been proven with accurate prediction and could decrease drug development burden, inform clinical design and potentially waive clinical studies. Formulation strategies and careful clinical design could help mitigate the pH-mediated DDI to avoid more clinical studies and label restrictions, ultimately benefiting the patient. In this review paper, a detailed introduction on biorelevant dissolution testing, preclinical and clinical study requirement and PBPK modeling approaches to assess ARA-DDI are described. An improved decision tree for pH-mediated DDI is proposed. Potential mitigations including clinical or formulation strategies are discussed.

Enzyme-mediated drug-drug interactions: a review of in vivo and in vitro methodologies, regulatory guidance, and translation to the clinic.

Enzyme-mediated pharmacokinetic drug-drug interactions can be caused by altered activity of drug metabolizing enzymes in the presence of a perpetrator drug, mostly via inhibition or induction. We identified a gap in the literature for a state-of-the art detailed overview assessing this type of DDI risk in the context of drug development. This manuscript discusses in vitro and in vivo methodologies employed during the drug discovery and development process to predict clinical enzyme-mediated DDIs, including the determination of clearance pathways, metabolic enzyme contribution, and the mechanisms and kinetics of enzyme inhibition and induction. We discuss regulatory guidance and highlight the utility of in silico physiologically-based pharmacokinetic modeling, an approach that continues to gain application and traction in support of regulatory filings. Looking to the future, we consider DDI risk assessment for targeted protein degraders, an emerging small molecule modality, which does not have recommended guidelines for DDI evaluation. Our goal in writing this report was to provide early-career researchers with a comprehensive view of the enzyme-mediated pharmacokinetic DDI landscape to aid their drug development efforts.

Crude protein oscillation in diets adequate and deficient in metabolizable protein: Effects on nutrient digestibility, nitrogen balance, plasma amino acids, and greenhouse gas emissions.

Reducing dietary CP is a well-established means to improve N use efficiency. Yet, few studies have considered if transient restrictions in dietary CP could reduce the environmental footprint of late-lactation cows. We hypothesized that the effects of CP feeding pattern on digestibility and environmental outputs would be amplified at lower dietary CP. We tested CP levels below and near predicted requirements (low protein [LP], 13.8%; high protein [HP], 15.5%) offered in 2 feeding patterns: where diets alternated ±1.8 percentage units CP every 2 d (oscillating [OF]) or remained static. Our study used a 2 × 2 factorial design with 16 mid- to late-lactation Holsteins (mean = 128, SD = 12 DIM), divided into rumen-cannulated (n = 8) and noncannulated subsets (n = 8). For each 28-d experimental period, we recorded feed intake and milk production and took samples of orts (1×/d) and milk (2×/d) for 4 d. For the cannulated subset, we measured and sampled from the total mass of feces and urine production and collected plasma 2×/d across 4 d. For the noncannulated subset, we sampled carbon dioxide and methane emissions 3×/d for 4 d. For each subset, we fit linear mixed models with fixed effects for CP level, CP feeding pattern, the interaction of CP level and CP feeding pattern, period, and a random effect for cow. For plasma and urinary urea-N, we conducted time series analysis. Contrary to our hypothesis, we found no evidence that dietary CP level and CP feeding pattern interacted to influence N balance, nutrient digestibility, or gas emissions. Results showed HP resulted in similar milk N but increased manure N, reducing N use efficiency (milk true protein N/intake N) relative to LP. For OF, urea-N in urine and plasma peaked 46 to 52 h after the first higher-CP phase feeding. Nutrient digestibility and gas emissions were similar across treatments, except CO2 production was greater for OF-HP. In summary, measured variables were minimally affected by dietary CP alternating ±1.8 percentage units every 48 h, even when average dietary CP was fed below predicted requirements (LP). Although our findings suggest that mid- to late-lactation cows are resilient to oscillation in dietary CP, oscillating CP neither reduced the environmental footprint by improving nutrient use efficiencies nor reduced the potential for direct and indirect greenhouse gas emissions.

Reducing dietary crude protein: Effects on digestibility, nitrogen balance, and blood metabolites in late-lactation Holstein cows.

Our objectives were to determine the effects of reducing dietary CP concentration on nutrient digestibility, rumen function, N balance, and serum AA concentration for dairy cows in late lactation. At the initiation of the experimental period, we stratified Holstein cows (n = 128; mean ± SD 224 ± 54 DIM) by parity and days pregnant (86 ± 25 d) and assigned them to 1 of 16 pens. For 3 wk, all cows received a covariate diet containing 16.9% CP (DM basis). For the subsequent 12 wk, we assigned pens to 1 of 4 treatments containing 16.2%, 14.4%, 13.4%, or 11.9% CP (DM basis) in a randomized complete block design. Diets were fed as a TMR once daily. To reduce dietary CP, we replaced soybean meal with soybean hulls in the concentrate mix (DM basis). Diet evaluations suggested that several EAA, especially His, limited productivity as dietary CP declined. Digestibility of DM and CP decreased linearly with dietary CP reduction. Digestibility of NDF and potentially digestible NDF tended to respond in a quadratic pattern with the greatest digestibility at intermediate treatments. The reduction in dietary CP did not affect ruminal pH, but ruminal ammonia-N and branched-chain VFA concentrations declined linearly. The concentration of milk urea-N and plasma urea-N, secretion of milk N, and excretions of fecal N, urinary N, urinary urea-N, and unaccounted N decreased linearly with the reduction in dietary CP concentration. Urinary N expressed as a percentage of N intake was unaffected by dietary CP. Serum concentrations of total essential AA and NEAA were unaffected by dietary CP concentration. However, the ratio of essential to NEAA decreased with decreasing dietary CP. Serum 3-methylhistidine concentration increased linearly with decreasing dietary CP concentration, indicating greater skeletal muscle breakdown. Although our trial confirmed that reducing dietary CP decreased absolute excretion of urinary N, diet evaluations suggested that milk protein production decreased as certain essential AA became increasingly limited. Thus, reduced-CP diets have the potential to lessen reactive-N outputs of late-lactation cows, but more research is needed to design diets that minimize deleterious effects on productivity.

Dynamic lactation responses to dietary crude protein oscillation in diets adequate and deficient in metabolizable protein in Holstein cows.

Limited research has examined the interaction between dietary crude protein (CP) level and CP feeding pattern. We tested CP level (low protein [LP], 13.8%; high protein [HP], 15.5% CP, dry matter [DM] basis) and CP feeding pattern (OF = oscillating, SF = static) using a 2 × 2 factorial in 16 mid- to late-lactation Holsteins (initially 128 ± 12 d in milk; mean ± SD). Cows ate total mixed rations formulated by exchanging soy hulls and ground corn with solvent soybean meal to keep constant ratios of neutral detergent fiber to starch (1.18:1), rumen-degradable protein to CP (0.61:1), and forage-to-concentrate (1.5:1) in DM. The OF treatments alternated diets every 48 h to vary CP above and below the mean CP level (OF-LP = 13.8% ± 1.8%; OF-HP = 15.5% ± 1.8% CP [DM basis]) whereas diets were constant in SF (SF-LP = 13.8%; SF-HP = 15.5% CP [DM basis]). In four 28-d periods, 8 rumen-cannulated and 8 noncannulated cows formed 2 Latin rectangles. On d 25 to 28 of each period, each cow's feed intake and milk production were recorded, and samples were taken of orts (1×/d) and milk (2×/d). We fit linear mixed models with fixed CP level, CP feeding pattern, and period effects, and a random intercept for cow, computing least squares means and standard errors. Neither CP level, CP feeding pattern, nor the interaction affected DM intake, feed efficiency, or production of milk, fat- and protein-corrected milk (FPCM), fat, true protein, or lactose. Milk urea-N (MUN) yield was lesser for LP. The LP and OF conditions decreased MUN concentration. The CP level tended to interact with CP feeding pattern so that milk protein concentration was greatest for OF-HP. The OF and LP conditions increased the ratio of true protein to MUN yield. Within OF, cosinor mixed models of selected variables showed that cows maintained production of FPCM across dietary changes, but MUN followed a wave-pattern at a 2-d delay relative to dietary changes. A tendency for lesser MUN with OF contradicted prior research and suggested potential differences in urea-N metabolism between OF and SF. Results showed that cows maintained production of economically-relevant components regardless of CP feeding pattern and CP level. Contrary to our hypothesis, the effects of 48-h oscillating CP were mostly consistent across CP levels, suggesting that productivity is resilient to patterned variation in dietary CP over time even when average CP supply is low (13.8% of DM) and despite 48 h restrictions at 12.2% CP.

Epigenetics in drug disposition & drug therapy: symposium report of the 24th North American meeting of the International Society for the Study of Xenobiotics (ISSX).

The 24th North American International Society for the Study of Xenobiotics (ISSX) meeting, held virtually from September 13 to 17, 2021, embraced the theme of "Broadening Our Horizons." This reinforces a key mission of ISSX: striving to share innovative science related to drug discovery and development. Session speakers and the ISSX New Investigators Group, which supports the scientific and professional development of student and early career ISSX members, elected to highlight the scientific content presented during the captivating session titled, "Epigenetics in Drug Disposition & Drug Therapy." The impact genetic variation has on drug response is well established; however, this session underscored the importance of investigating the role of epigenetics in drug disposition and drug discovery. Session speakers, Drs. Ning, McClay, and Lazarus, detailed mechanisms by which epigenetic players including long non-coding RNA (lncRNAs), microRNA (miRNAs), DNA methylation, and histone acetylation can alter the expression of genes involved in pharmacokinetics, pharmacodynamics, and toxicity. Dr. Ning detailed current knowledge about miRNAs and lncRNAs and the mechanisms by which they can affect the expression of drug metabolizing enzymes (DMEs) and nuclear receptors. Dr. Lazarus discussed the potential role of miRNAs on UDP-glucuronosyltransferase (UGT) expression and activity. Dr. McClay provided evidence that aging alters methylation and acetylation of DMEs in the liver, affecting gene expression and activity. These topics, compiled by the symposium organizers, presenters, and the ISSX New Investigators Group, are herein discussed, along with exciting future perspectives for epigenetics in drug disposition and drug discovery research.

Complex Cytochrome P450 Kinetics Due to Multisubstrate Binding and Sequential Metabolism. Part 1. Theoretical Considerations.

Complexities in P450-mediated metabolism kinetics include multisubstrate binding, multiple-product formation, and sequential metabolism. Saturation curves and intrinsic clearances were simulated for single-substrate and multisubstrate models using derived velocity equations and numerical solutions of ordinary differential equations (ODEs). Multisubstrate models focused on sigmoidal kinetics because of their dramatic impact on clearance predictions. These models were combined with multiple-product formation and sequential metabolism, and simulations were performed with random error. Use of single-substrate models to characterize multisubstrate data can result in inaccurate kinetic parameters and poor clearance predictions. Comparing results for use of standard velocity equations with ODEs clearly shows that ODEs are more versatile and provide better parameter estimates. It would be difficult to derive concentration-velocity relationships for complex models, but these relationships can be easily modeled using numerical methods and ODEs. SIGNIFICANCE STATEMENT: The impact of multisubstrate binding, multiple-product formation, and sequential metabolism on the P450 kinetics was investigated. Numerical methods are capable of characterizing complicated P450 kinetics.

Complex Cytochrome P450 Kinetics Due to Multisubstrate Binding and Sequential Metabolism. Part 2. Modeling of Experimental Data.

Three CYP3A4 substrates, midazolam, ticlopidine, and diazepam, display non-Michaelis-Menten kinetics, form multiple primary metabolites, and are sequentially metabolized to secondary metabolites. We generated saturation curves for these compounds and analyzed the resulting datasets using a number of single-substrate and multisubstrate binding models. These models were parameterized using rate equations and numerical solutions of the ordinary differential equations. Multisubstrate binding models provided results superior to single-substrate models, and simultaneous modeling of multiple metabolites provided better results than fitting the individual datasets independently. Although midazolam datasets could be represented using standard two-substrate models, more complex models that include explicit enzyme-product complexes were needed to model the datasets for ticlopidine and diazepam. In vivo clearance predictions improved markedly with the use of in vitro parameters from the complex models versus the Michaelis-Menten equation. The results highlight the need to use sufficiently complex kinetic schemes instead of the Michaelis-Menten equation to generate accurate kinetic parameters. SIGNIFICANCE STATEMENT: The metabolism of midazolam, ticlopidine, and diazepam by CYP3A4 results in multiple metabolites and sequential metabolism. This study evaluates the use of rate equations and numerical methods to characterize the in vitro enzyme kinetics. Use of complex cytochrome P450 kinetic models is necessary to obtain accurate parameter estimates for predicting in vivo disposition.

Aldehyde Oxidase Contributes to All-Trans-Retinoic Acid Biosynthesis in Human Liver.

All-trans-retinoic acid (atRA) is a critical endogenous signaling molecule. atRA is predominantly synthesized from retinaldehyde by aldehyde dehydrogenase 1A1 (ALDH1A1), but aldehyde oxidase (AOX) may also contribute to atRA biosynthesis. The goal of this study was to test the hypothesis that AOX contributes significantly to atRA formation in human liver. Human recombinant AOX formed atRA from retinaldehyde (Km ∼1.5 ± 0.4 µM; kcat ∼3.6 ± 2.0 minute-1). In human liver S9 fractions (HLS9), atRA formation was observed in the absence of NAD+, suggesting AOX contribution to atRA formation. In the presence of NAD+, Eadie-Hofstee plots of atRA formation in HLS9 indicated that two enzymes contributed to atRA formation. The two enzymes were identified as AOX and ALDH1A1 based on inhibition of atRA formation by AOX inhibitor hydralazine (20%-50% inhibition) and ALDH1A1 inhibitor WIN18,446 (50%-80%inhibition). The expression of AOX in HLS9 was 9.4-24 pmol mg-1 S9 protein, whereas ALDH1A1 expression was 156-285 pmol mg-1 S9 protein measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) quantification of signature peptides. The formation velocity of atRA in the presence of NAD+ correlated significantly with the expression of ALDH1A1 and AOX protein. Taken together, the data show that both AOX and ALDH1A1 contribute to atRA biosynthesis in the human liver, with ALDH1A1 being the high-affinity, low-capacity enzyme and AOX being the low-affinity, high-capacity enzyme. The results suggest that in the case of ALDH1A dysfunction or excess vitamin A, AOX may play an important role in regulating hepatic vitamin A homeostasis and that inhibition of AOX may alter atRA biosynthesis and signaling. SIGNIFICANCE STATEMENT: This study provides direct evidence to show that human AOX converts retinaldehyde to atRA and contributes to hepatic atRA biosynthesis. The finding that AOX may be responsible for 20%-50% of overall hepatic atRA formation suggests that alterations in AOX activity via drug-drug interactions, genetic polymorphisms, or disease states may impact hepatic atRA concentrations and signaling and alter vitamin A homeostasis.

Treatment and preliminary outcomes of 150 acute care patients with COVID-19 in a rural health system in the Dakotas.

The majority of available US-published reports present populations with community spread in urban areas. The objective of this report is to describe a rural healthcare system's utilisation of therapeutic options available to treat Coronavirus Disease 2019 (COVID-19) and subsequent patient outcomes. A total of 150 patients were treated for COVID-19 at three hospitals in the Dakotas from 21 March 2020 to 30 April 2020. The most common pharmacological treatment regimens administered were zinc, hydroxychloroquine plus azithromycin and convalescent plasma. Adjunctive treatments included therapeutic anticoagulation, tocilizumab and corticosteroids. As of 1 June 2020, 127 patients have survived to hospital discharge, 12 patients remain hospitalised and 11 patients have expired. The efficacy of hydroxychloroquine and azithromycin use has yet to be determined but was not without risks of corrected QT interval prolongation and arrhythmias in our cohort. We did not appreciate any adverse effects that appeared related to tocilizumab or convalescent plasma administration in those patient subsets. These findings may provide insight into disease severity and treatment options in the rural setting with limited resources to participate in clinical trials and encourage larger comparative studies evaluating treatment efficacy.

Short communication: Effect of glucose infusion dose and stage of lactation on glucose tolerance test kinetics in lactating dairy cows.

The objective for this study was to determine the effect of glucose dose and days following peak milk yield on plasma glucose, serum insulin, and plasma nonesterified fatty acids (NEFA) kinetics during an intravenous glucose tolerance test (IVGTT) in lactating dairy cattle. Six lactating Holstein dairy cows (3 primiparous and 3 multiparous) were assigned to 2 squares and received 0.092, 0.15, or 0.3 g of glucose/kg of body weight (BW) during an IVGTT at 74 and 221 d in milk (DIM), representing early (post-peak) lactation and mid lactation, respectively. Treatments were applied in a replicated Latin square design using contiguous 7-d periods within each stage of lactation. Milk production and dry matter intake were determined daily during the first 6 d of each period. The IVGTT was performed on d 7. For the IVGTT, cows were prepared with indwelling catheters in each jugular vein, and blood samples were collected at -15, -10, 5, 10, 15, 20, 30, 45, 60, 90, and 120 min relative to the glucose infusion. Samples were analyzed for plasma glucose, serum insulin, and plasma NEFA concentrations. Increasing the glucose dose during the IVGTT increased plasma glucose area under the curve (AUC), decreased glucose half-life, and increased maximal plasma glucose concentrations in plasma during the IVGTT. Greater glucose dose during the IVGTT elevated serum insulin AUC and increased nadir NEFA concentrations. Maximal plasma glucose concentration during the IVGTT was lower, whereas maximum NEFA concentration, NEFA AUC, and NEFA clearance rate were greater at 74 than at 221 DIM. Only glucose half-life was responsive to stage of lactation × glucose dose effects during the IVGTT, and the decrease in glucose half-life with increasing glucose dose was greater at 74 than at 221 DIM. Glucose AUC was greater and NEFA AUC lower for cows at 74 than at 221 DIM. For the doses tested, a glucose dose greater than 0.092 g/kg of BW resulted in peak blood glucose concentration that exceeded the previously reported renal glucose excretion threshold of 8.3 mM. There is a need for accompanying data to determine if this is the case for the glucose doses evaluated in this experiment. Based on maximal peak glucose concentrations and effects on glucose half-life, we identify 0.092 g of glucose/kg of BW (0.46 g/kg of metabolic body weight) as the preferred dose for the IVGTT for cows at 74 and 221 DIM in this study.

Time Course of Aldehyde Oxidase and Why It Is Nonlinear.

Many promising drug candidates metabolized by aldehyde oxidase (AOX) fail during clinical trial owing to underestimation of their clearance. AOX is species-specific, which makes traditional allometric studies a poor choice for estimating human clearance. Other studies have suggested using half-life calculated by measuring substrate depletion to measure clearance. In this study, we proposed using numerical fitting to enzymatic pathways other than Michaelis-Menten (MM) to avoid missing the initial high turnover rate of product formation. Here, product formation over a 240-minute time course of six AOX substrates-O6-benzylguanine, N-(2-dimethylamino)ethyl)acridine-4-carboxamide, zaleplon, phthalazine, BIBX1382 [N8-(3-Chloro-4-fluorophenyl)-N2-(1-methyl-4-piperidinyl)-pyrimido[5,4-d]pyrimidine-2,8-diamine dihydrochloride], and zoniporide-have been provided to illustrate enzyme deactivation over time to help better understand why MM kinetics sometimes leads to underestimation of rate constants. Based on the data provided in this article, the total velocity for substrates becomes slower than the initial velocity by 3.1-, 6.5-, 2.9-, 32.2-, 2.7-, and 0.2-fold, respectively, in human expressed purified enzyme, whereas the K m remains constant. Also, our studies on the role of reactive oxygen species (ROS), such as superoxide and hydrogen peroxide, show that ROS did not significantly alter the change in enzyme activity over time. Providing a new electron acceptor, 5-nitroquinoline, did, however, alter the change in rate over time for mumerous compounds. The data also illustrate the difficulties in using substrate disappearance to estimate intrinsic clearance.

Short communication: Decreasing the dietary ratio of n-6 to n-3 fatty acids increases the n-3 concentration of peripheral blood mononuclear cells in weaned Holstein heifer calves.

Utilization of nutrients to improve overall heifer health is of interest because of the importance of replacement heifers to the dairy industry. The objective of our study was to compare the effect of supplementation of dietary n-3 and n-6 fatty acids (FA) on FA concentrations in peripheral blood mononuclear cells (PBMC) of Holstein calves. Twenty-seven Holstein heifer calves (107 ± 2.6 d of age; 142.6 ± 6.5 kg of body weight) from the university research and teaching herd were randomly assigned to a common TMR supplemented with 1 of 3 treatments: Ca salts of flaxseed FA (Virtus Nutrition, Corcoran, CA) containing 35% 18:3 n-3 (N3), Ca salts of soybean FA (Virtus Nutrition) containing 50% 18:2 n-6 (N6), or a 50:50 mix of N3 and N6. Treatments were supplemented with FA at 4% of dietary dry matter and fed for 30 d. Feed intake was recorded daily, and body weight, wither height, and body condition score were measured weekly throughout the study. On d 28 heifers were vaccinated with a Pasteurella vaccine and the temperature response to the vaccine was recorded. Blood was collected on d 0 and 28 for PBMC isolation. After total lipid extraction and FA methyl ester preparation, FA composition of PBMC was measured. We observed no effect of treatment on body weight gain, body condition score change, or wither height change. Heifers receiving the N3 diet had a lower temperature response to Pasteurella challenge compared with both the mix and N6 diets. Heifers consuming the N3 diet had a greater content of total n-3 FA, α-linolenic acid, and eicosapentaenoic acid in PBMC compared with heifers fed the N6 and mix diets. Heifers receiving the N3 diet also had a lower content of total n-6 FA, linoleic acid, and arachidonic acid in PBMC than heifers fed the N6 and mix diets. In conclusion, our study determined that feeding weaned female Holstein heifers a diet high in n-3 FA increased concentrations of n-3 FA in PBMC.

Examination of factors for use as potential predictors of human enteric pathogen survival in soil.

AIMS: Three soils that varied in their physicochemical characteristics and microbial diversity were inoculated with Escherichia coli O157:H7 and Salmonella to determine the relative impact of abiotic and biotic factors on the pathogens' survival when the soil was held at 25°C. METHODS AND RESULTS: Three soils that were classified as having low, medium and high microbial diversity were divided into two batches for adjustment to 20% of water-holding capacity and to 40% of water-holding capacity. Soils were inoculated with both green fluorescent-labelled E. coli O157:H7 and red fluorescent-labelled Salmonella (5 log CFU g(-1) dry weight) and held at 25°C. Pathogens inoculated into an acidic soil died off within 9 weeks, whereas they were still detected in the other two soils by enrichment culture after 18 weeks. Moisture did not affect inactivation of E. coli O157:H7, but did affect Salmonella inactivation in soil having the greatest organic load and microbial diversity. Using multiple linear regression analysis, 98.7% of the variability in the inactivation rate for E. coli O157:H7 was explained by a model that included the variables of initial pH and electrical conductivity. Salmonella's inactivation rate was predicted by a model that included pH and initial cell numbers of copiotrophic and oligotrophic bacteria. CONCLUSION: This study provided evidence of specific properties that impact inactivation of E. coli O157:H7 and Salmonella in soils at 25°C. SIGNIFICANCE AND IMPACT OF THE STUDY: Identification of factors influential in the die-off of enteric pathogens will assist in developing guidelines for safe intervals between field contamination events and planting or harvesting of fresh-cut produce crops.

Observations and properties of the first laboratory fusion experiment to exceed a target gain of unity.

An indirect-drive inertial fusion experiment on the National Ignition Facility was driven using 2.05 MJ of laser light at a wavelength of 351 nm and produced 3.1±0.16 MJ of total fusion yield, producing a target gain G=1.5±0.1 exceeding unity for the first time in a laboratory experiment [Phys. Rev. E 109, 025204 (2024)10.1103/PhysRevE.109.025204]. Herein we describe the experimental evidence for the increased drive on the capsule using additional laser energy and control over known degradation mechanisms, which are critical to achieving high performance. Improved fuel compression relative to previous megajoule-yield experiments is observed. Novel signatures of the ignition and burn propagation to high yield can now be studied in the laboratory for the first time.

Surface roughness dominated pinning mechanism of magnetic vortices in soft ferromagnetic films.

Although pinning of domain walls in ferromagnets is ubiquitous, the absence of an appropriate characterization tool has limited the ability to correlate the physical and magnetic microstructures of ferromagnetic films with specific pinning mechanisms. Here, we show that the pinning of a magnetic vortex, the simplest possible domain structure in soft ferromagnets, is strongly correlated with surface roughness, and we make a quantitative comparison of the pinning energy and spatial range in films of various thickness. The results demonstrate that thickness fluctuations on the lateral length scale of the vortex core diameter, i.e., an effective roughness at a specific length scale, provides the dominant pinning mechanism. We argue that this mechanism will be important in virtually any soft ferromagnetic film.

Enzyme Kinetics, Pharmacokinetics, and Inhibition of Aldehyde Oxidase.

Aldehyde oxidase (AO) has emerged as an important drug metabolizing enzyme over the last decade. Several compounds have failed in the clinic because the clearance or toxicity was underestimated by preclinical species. Human AO is much more active than rodent AO, and dogs do not have functional AO. Metabolic products from AO-catalyzed oxidation are generally nonreactive and often they have much lower solubility. AO metabolism is not limited to oxidation as AO can also catalyze reduction of oxygen and nitrite. Reduction of oxygen leads to the reactive oxygen species (ROS) superoxide radical anion and hydrogen peroxide. Reduction of nitrite leads to the formation of nitric oxide with potential pharmacological implications. AO is also reported to catalyze the reductive metabolism of nitro-compounds, N-oxides, sulfoxides, isoxazoles, isothiazoles, nitrite, and hydroxamic acids. These reductive transformations may cause toxicity due to the formation of reactive metabolites. Moreover, the inhibition kinetics are complex, and multiple probe substrates should be used when assessing the potential for DDIs. Finally, AO appears to be amenable to computational predictions of both regioselectivity and rates of reaction, which holds promise for virtual screening.

Use of a mixture of bacteriophages for biological control of Salmonella enterica strains in compost.

Bacteriophages specific to Salmonella strains were isolated from sewage effluent and characterized. A five-strain bacteriophage mixture was applied to dairy manure compost inoculated with Salmonella enterica serotype Typhimurium. Bacteriophage treatment resulted in a greater than 2-log-unit reduction of Salmonella within 4 h at all moisture levels compared to the controls.

Microbiological analysis of composts produced on South Carolina poultry farms.

AIMS: The purpose of this study was to determine whether the methods used in compost operations of small and medium-sized poultry farms resulted in the production of an amendment free of foodborne pathogens. METHODS AND RESULTS: Nine compost heaps on five South Carolina poultry farms were surveyed at different stages of the composting process. Compost samples were analysed for coliforms and enriched for Escherichia coli O157:H7, Salmonella spp. and Listeria monocytogenes. The waste materials and composting practices differed among the surveyed farms. On two farms, new materials were added to heaps that had previously completed the active composting phase. Five compost heaps did not reach an internal temperature of 55 degrees C, and c. 62% of all internal samples in the first composting phase contained moisture contents <40%. Escherichia coli was detected in 63% of the surface samples (n = 38) and 9.8% of the internal samples (n = 82) from the first composting phase, as compared with 16.7% of the surface samples (n = 12) and 0% internal samples (n = 24) from the second composting phase. Salmonella was detected in 26 and 6.1% of all surface and internal samples collected from heaps in the first composting phase, respectively, but was absent in all compost samples undergoing a second composting phase. The predominant Salmonella serotypes were Thompson, Montevideo and Anatum. Neither E. coli O157:H7 nor L. monocytogenes was detected in any of the samples. CONCLUSIONS: Our results indicate that the conditions at the compost surface are suitable for pathogen survival, and the complete composting process can result in the elimination of pathogens in poultry wastes. SIGNIFICANCE AND IMPACT OF THE STUDY: This research provides information regarding the effectiveness of the composting practices and microbiological quality of poultry compost produced by small- and medium-sized farms. Ensuring the safety of compost that may be applied to soils should be an integral part of preharvest food safety programme.