Kirkwood-Buff integrals: From fluctuations in finite volumes to the thermodynamic limit.

The Kirkwood-Buff theory is a cornerstone of the statistical mechanics of liquids and solutions. It relates volume integrals over the radial distribution function, so-called Kirkwood-Buff integrals (KBIs), to particle number fluctuations and thereby to various macroscopic thermodynamic quantities such as the isothermal compressibility and partial molar volumes. Recently, the field has seen a strong revival with breakthroughs in the numerical computation of KBIs and applications to complex systems such as bio-molecules. One of the main emergent results is the possibility to use the finite volume KBIs as a tool to access finite volume thermodynamic quantities. The purpose of this Perspective is to shed new light on the latest developments and discuss future avenues.

Advanced flow MRI: emerging techniques and applications.

Magnetic resonance imaging (MRI) techniques provide non-invasive and non-ionising methods for the highly accurate anatomical depiction of the heart and vessels throughout the cardiac cycle. In addition, the intrinsic sensitivity of MRI to motion offers the unique ability to acquire spatially registered blood flow simultaneously with the morphological data, within a single measurement. In clinical routine, flow MRI is typically accomplished using methods that resolve two spatial dimensions in individual planes and encode the time-resolved velocity in one principal direction, typically oriented perpendicular to the two-dimensional (2D) section. This review describes recently developed advanced MRI flow techniques, which allow for more comprehensive evaluation of blood flow characteristics, such as real-time flow imaging, 2D multiple-venc phase contrast MRI, four-dimensional (4D) flow MRI, quantification of complex haemodynamic properties, and highly accelerated flow imaging. Emerging techniques and novel applications are explored. In addition, applications of these new techniques for the improved evaluation of cardiovascular (aorta, pulmonary arteries, congenital heart disease, atrial fibrillation, coronary arteries) as well as cerebrovascular disease (intra-cranial arteries and veins) are presented.

Effects of silver nanoparticles on microbial growth dynamics.

AIMS: Engineered metal nanoparticles are increasingly used in consumer products, in part as additives that exhibit advantageous antimicrobial properties. Conventional nanoparticle susceptibility testing is based largely on determination of nontemporal growth profiles such as measurements of inhibition zones in common agar diffusion tests, counting of colony-forming units, or endpoint or regular-interval growth determination via optical density measurements. For better evaluation of the dynamic effects from exposure to nanoparticles, a cultivation-based assay was established in a 96-well format and adapted for time-resolved testing of the effects of nanoparticles on micro-organisms. METHODS AND RESULTS: The modified assay allowed simultaneous cultivation and on-line analysis of microbial growth inhibition. The automated high-throughput assay combined continuous monitoring of microbial growth with the analysis of many replicates and was applied to Cupriavidus necator H16 test organisms to study the antimicrobial effects of spherical silver [Ag(0)] nanoparticles (primary particle size distribution D90 < 15 nm). Ag(0) concentrations above 80 µg ml(-1) resulted in complete and irreversible inhibition of microbial growth, whereas extended lag phases and partial growth inhibition were observed at Ag(0) concentrations between 20 and 80 µg ml(-1) . Addition of Ag(0) nanoparticles at different growth stages led to either complete inhibition (addition of 40 µg ml(-1) Ag(0) from 0 h to 6 h) or resulted in full recovery (40 µg ml(-1) Ag(0) addition ≥9 h). CONCLUSIONS: Contrary to the expected results, our data indicate growth stimulation of C. necator at certain Ag(0) nanoparticle concentrations, as well as varying susceptibility to nanoparticles at different growth stages. SIGNIFICANCE AND IMPACT OF THE STUDY: These results underscore the need for time-resolved analyses of microbial growth inhibition by Ag(0) nanoparticles. Due to the versatility of the technique, the assay will likely complement existing microbiological methods for cultivation and diagnostics of microbes, in addition to tests of other antimicrobial nanoparticles.

Growth inhibition of Cronobacter spp. strains in reconstituted powdered infant formula acidified with organic acids supported by natural stomach acidity.

Cronobacter is associated with outbreaks of rare, but life-threatening cases of meningitis, necrotizing enterocolitis, and sepsis in newborns. This study was conducted to determine the effect of organic acids on growth of Cronobacter in laboratory medium and reconstituted powdered infant formula (PIF) as well as the bacteriostatic effect of slightly acidified infant formula when combined with neonatal gastric acidity. Inhibitory effect of seven organic acids on four acid sensitive Cronobacter strains was determined in laboratory medium with broth dilution method at pH 5.0, 5.5 and 6.0. Acetic, butyric and propionic acids were most inhibitive against Cronobacter in the laboratory medium. The killing effect of these three acids was partially buffered in reconstituted PIF. Under neonatal gastric acid condition of pH 5.0, the slightly acidified formula which did not exert inhibition effect solely reduced significantly the Cronobacter populations. A synergistic effect of formula moderately acidified with organic acid combined with the physiological infant gastric acid was visible in preventing the rapid growth of Cronobacter in neonatal stomach. The study contributed to a better understanding of the inhibitory effect of organic acids on Cronobacter growth in different matrixes and provided new ideas in terms of controlling bacteria colonization and translocation by acidified formula.

Cerebrovascular super-resolution 4D Flow MRI - Sequential combination of resolution enhancement by deep learning and physics-informed image processing to non-invasively quantify intracranial velocity, flow, and relative pressure.

The development of cerebrovascular disease is tightly coupled to regional changes in intracranial flow and relative pressure. Image-based assessment using phase contrast magnetic resonance imaging has particular promise for non-invasive full-field mapping of cerebrovascular hemodynamics. However, estimations are complicated by the narrow and tortuous intracranial vasculature, with accurate image-based quantification directly dependent on sufficient spatial resolution. Further, extended scan times are required for high-resolution acquisitions, and most clinical acquisitions are performed at comparably low resolution (>1 mm) where biases have been observed with regard to the quantification of both flow and relative pressure. The aim of our study was to develop an approach for quantitative intracranial super-resolution 4D Flow MRI, with effective resolution enhancement achieved by a dedicated deep residual network, and with accurate quantification of functional relative pressures achieved by subsequent physics-informed image processing. To achieve this, our two-step approach was trained and validated in a patient-specific in-silico cohort, showing good accuracy in estimating velocity (relative error: 15.0 ± 0.1%, mean absolute error (MAE): 0.07 ± 0.06 m/s, and cosine similarity: 0.99 ± 0.06 at peak velocity) and flow (relative error: 6.6 ± 4.7%, root mean square error (RMSE): 0.56 mL/s at peak flow), and with the coupled physics-informed image analysis allowing for maintained recovery of functional relative pressure throughout the circle of Willis (relative error: 11.0 ± 7.3%, RMSE: 0.3 ± 0.2 mmHg). Furthermore, the quantitative super-resolution approach is applied to an in-vivo volunteer cohort, effectively generating intracranial flow images at <0.5 mm resolution and showing reduced low-resolution bias in relative pressure estimation. Our work thus presents a promising two-step approach to non-invasively quantify cerebrovascular hemodynamics, being applicable to dedicated clinical cohorts in the future.

Agave fructans enhance the effects of fermented milk products on obesity biomarkers: a randomised trial.

Dysbiosis has been implicated in childhood obesity. Oral intake of fermented milk containing Lacticaseibacillus casei strain Shirota preserves gut microbiota (GM) diversity in children and adults. This study was a double-blind trial involving 37 overweight or obese children aged 6-10 years. Children were followed over a 6-week intervention period in which they received different fermented milk products containing L. casei Shirota: 10 in the first group received just L. casei Shirota; 13 received L. casei Shirota with 3 g/day of inulin (L. casei+inulin); and 14 received L. casei Shirota with 3 g/day of fructans from Agave salmiana (L. casei+fructans). Principal component analysis showed the relationship between microbial abundance, GM metabolites, and other obesity-related markers. Supplementation with probiotics and synbiotics improved the HDL-cholesterol levels of overweight and obese children, although no changes in body composition were detected. We observed an increase in butyrate or propionate concentrations in the L. casei+fructans group compared to the end of the intervention (P<0.03). A diminished level of ANGPTL4 within the L. casei+fructans group (P=0.04) was also found, but no differences when lipopolysaccharide-binding protein was evaluated. The FFAR2+ cell frequency decreased between baseline and at the end of 6-week intervention in L. casei+inulin (P=0.02) and L. casei+fructans groups (P=0.04). In contrast, the percentage of CD14+FFAR3+ frequency increased in the same groups (P=0.04). The L. casei Shirota with inulin or fructans modulates GM, which improves the lipid profile and changes at a molecular level, such as expression of FFAR3 and FFAR2, ANGPTL4, propionate, and butyrate. It, therefore, could be considered an interesting therapeutic possibility for treating childhood overweight and obesity. The study was registered at ClinicalTrials.gov (ID: NCT05423015).

Unexpected stability of Bacteroidetes and Firmicutes communities in laboratory biogas reactors fed with different defined substrates.

In the present study, bacterial communities in 200-liter biogas reactors containing liquid manure consecutively fed with casein, starch, and cream were investigated over a period of up to 33 days. A 16S rRNA gene clone library identified Bacteroidetes and Firmicutes as the most abundant bacterial groups in the starting material, at 58.9% and 30.1% of sequences, respectively. The community development of both groups was monitored by real-time PCR and single-strand conformation polymorphism (SSCP) analysis. The Firmicutes and Bacteroidetes communities were unexpectedly stable and hardly influenced by batch-feeding events. The continuous feeding of starch led to community shifts that nevertheless contributed to a stable reactor performance. A longer starving period and a change in the pH value resulted in further community shifts within the Bacteroidetes but did not influence the Firmicutes. Predominant DNA bands from SSCP gels were cloned and sequenced. Sequences related to Peptococcaceae, Cytophagales, and Petrimonas sulfuriphila were found in all samples from all experiments. Real-time PCR demonstrated the abundance of members of the phylum Bacteroidetes and also reflected changes in gene copy numbers in conjunction with a changing pH value and acetate accumulation.

Fully automated classification of HARDI in vivo data using a support vector machine.

The purpose of this study is the classification of high angular resolution diffusion imaging (HARDI) in vivo data using a model-free approach. This is achieved by using a Support Vector Machine (SVM) algorithm taken from the field of supervised statistical learning. Six classes of image components are determined: grey matter, parallel neuronal fibre bundles in white matter, crossing neuronal fibre bundles in white matter, partial volume between white and grey matter, background noise and cerebrospinal fluid. The SVM requires properties derived from the data as input, the so called feature vector, which should be rotation invariant. For our application we derive such a description from the spherical harmonic decomposition of the HARDI signal. With this information the SVM is trained in order to find the function for separating the classes. The SVM is systematically tested with simulated data and then applied to six in vivo data sets. This new approach is data-driven and enables fully automatic HARDI data segmentation without employing a T1 MPRAGE scan and subjective expert intervention. This was demonstrated on five test in vivo data sets giving robust results. The segmentation results could be used as a priori knowledge for increasing the performance of fibre tracking as well as for other clinical and diagnostic applications of diffusion weighted imaging (DWI).

Screening, plant growth promotion and root colonization pattern of two rhizobacteria (Pseudomonas fluorescens Ps006 and Bacillus amyloliquefaciens Bs006) on banana cv. Williams (Musa acuminata Colla).

Banana is the second largest export crop in Colombia. To meet the demand of international markets, high amounts of chemical fertilizers are required, which represent high costs and can be hazardous to the environment. Plant growth promoting rhizobacteria (PGPR) can, at least partially, replace chemical fertilizers. In this paper, we evaluated the effect of nine PGPR of the genera Bacillus and Pseudomonas on banana growth. Banana seedlings were produced through tissue culture and acclimatized in the greenhouse core. Plants were inoculated with the rhizobacteria and growth parameters (plant height, leaf number, leaf area, pseudostem thickness, root and shoot fresh weight, root and shoot dry weight) were assessed after 55 days. The two best performing PGPR, Bs006 and Ps006 previously identified as Bacillus amyloliquefaciens and Pseudomonas fluorescens, respectively, promoted banana growth similarly or even slightly superior to 100% chemical fertilization, and were selected for further characterization of root colonization by both eletron microscopy and confocal microscopy of fluorescence in situ hybridization (FISH)-stained root tissues. Both P. fluorescens Ps006 and B. amyloquifaciens Bs006 showed ability to colonize banana roots, but Bs006 appeared faster than Ps006 in the colonization dynamics. This work demonstrated that inoculation of rhizobacteria Bacillus amyloliquefaciens Bs006 and Pseudomonas fluorescens Ps006 could partially replace the chemical fertilization of tissue cultured banana plants, and therefore could be used for the formulation of a new biofertilizer.

Connecting and merging fibres: pathway extraction by combining probability maps.

Probability mapping of connectivity is a powerful tool to determine the fibre structure of white matter in the brain. Probability maps are related to the degree of connectivity to a chosen seed area. In many applications, however, it is necessary to isolate a fibre bundle that connects two areas. A frequently suggested solution is to select curves, which pass only through two or more areas. This is very inefficient, especially for long-distance pathways and small areas. In this paper, a novel probability-based method is presented that is capable of extracting neuronal pathways defined by two seed points. A Monte Carlo simulation based tracking method, similar to the Probabilistic Index of Connectivity (PICo) approach, was extended to preserve the directional information of the main fibre bundles passing a voxel. By combining two of these extended visiting maps arising from different seed points, two independent parameters are determined for each voxel: the first quantifies the uncertainty that a voxel is connected to both seed points; the second represents the directional information and estimates the proportion of fibres running in the direction of the other seed point (connecting fibre) or face a third area (merging fibre). Both parameters are used to calculate the probability that a voxel is part of the bundle connecting both seed points. The performance and limitations of this DTI-based method are demonstrated using simulations as well as in vivo measurements.

Limit cycles in the presence of convection: a traveling wave analysis.

We consider a diffusion model with limit cycle reaction functions. In an unbounded domain, diffusion spreads the pattern outwards from the source. Convection adds instability to the reaction-diffusion system. The result of this instability is a readiness to create a pattern. We choose the Lambda-Omega reaction functions for their simple limit cycle. We carry out a transformation of the dependent variables into polar form. From this we consider the initiation of the pattern to approximate a traveling wave. We carry out numerical experiments to test our analysis. These confirm the premise of the analysis, that the initiation can be modeled by a traveling wave. Furthermore, the analysis produces a good estimate of the numerical results. Most significantly, we confirm that the pattern consists of two different types.

Genetic impacts of using female-sorted semen in commercial and nucleus herds.

This study was designed to investigate the genetic effects of using sorted semen in a dairy cattle population. Progress was monitored in elite and commercial animals over 20 yr of selection. To study the genetic impact of using sorted semen in commercial herds, a scenario was evaluated in which female-sorted semen was available to commercial herds. Second, to study the genetic impact of using sorted semen in nucleus herds, scenarios were simulated in which female-sorted semen was used only in a centralized nucleus herd, in which multiple ovulation and embryo transfer (MOET) took place. Because of the additional advantage of marker-assisted selection when sorted semen was used in nucleus herds, a second scenario was simulated in which both sorted semen and marker-assisted selection were implemented. In the scenario in which female-sorted semen was used in commercial herds, a large genetic advantage was observed early in commercial cows. The average superiority in first-lactation cows exceeded 30% in yr 11, relative to a base scheme with regular semen, but continued to decrease until it reached 9% in yr 20. The increased selection intensity in commercial cows contributed to the genetic merit of future cows (cow-to-cow contribution), but the contribution of the nucleus grew over time and gradually marginalized the cow-to-cow contribution. The genetic advantage of gender control in MOET schemes was minimal except when marker-assisted selection was also available. Two factors that affected the contribution of marker-assisted selection were studied: 1) within- vs. across-family selection of donors, and 2) the number of loci in the quantitative trait locus component. Schemes that selected donors regardless of their family structure were superior, and the quantitative trait locus component with more loci increased the effectiveness of sorted semen. Finally, we studied a reduced MOET scheme in which the number of harvested females was reduced from 42 to 25/yr. The reduced scheme in combination with female-sorted semen was not found to be genetically inferior to the large scheme in combination with regular semen.

Turing pattern outside of the Turing domain.

There are two simple solutions to reaction-diffusion systems with limit-cycle reaction kinetics, producing oscillatory behaviour. The reaction parameter mu gives rise to a 'space-invariant' solution, and mu versus the ratio of the diffusion coefficients gives rise to a 'time-invariant' solution. We consider the case where both solution types may be possible. This leads to a refinement of the Turing model of pattern formation. We add convection to the system and investigate its effect. More complex solutions arise that appear to combine the two simple solutions. The convective system sheds light on the underlying behaviour of the diffusive system.

In Vivo Assessment of the Impact of Regional Intracranial Atherosclerotic Lesions on Brain Arterial 3D Hemodynamics.

BACKGROUND AND PURPOSE: Intracranial atherosclerosis induces hemodynamic disturbance, which is not well-characterized, particularly in cerebral flow redistribution. We aimed to characterize the impact of regional stenotic lesions on intracranial hemodynamics by using 4D flow MR imaging. MATERIALS AND METHODS: 4D flow MR imaging was performed in 22 symptomatic patients (mean age, 68.4 ± 14.2 years) with intracranial stenosis (ICA, n = 7; MCA, n = 9; basilar artery, n = 6) and 10 age-appropriate healthy volunteers (mean age, 60.7 ± 8.1 years). 3D blood flow patterns were visualized by using time-integrated pathlines. Blood flow and peak velocity asymmetry indices were compared between patients and healthy volunteers in 4 prespecified arteries: ICAs, MCAs, and anterior/posterior cerebral arteries. RESULTS: 3D blood flow pathlines demonstrated flow redistribution across cerebral arteries in patients with unilateral intracranial stenosis. For patients with ICA stenosis compared with healthy volunteers, significantly lower flow and peak velocities were identified in the ipsilateral ICA (P = .001 and P = .001) and MCA (P < .001 and P = .001), but higher flow, in the ipsilateral PCA (P < .001). For patients with MCA stenosis, significantly lower flow and peak velocities were observed in the ipsilateral ICA (P = .009 and P = .045) and MCA (P < .001 and P = .005), but significantly higher flow was found in the ipsilateral posterior cerebral artery (P = .014) and anterior cerebral artery (P = .006). The asymmetry indices were not significantly different between patients with basilar artery stenosis and the healthy volunteers. CONCLUSIONS: Regional intracranial atherosclerotic lesions not only alter distal arterial flow but also significantly affect ipsilateral collateral arterial hemodynamics.

A systematic investigation of the rate laws valid in intracellular environments.

Recently there has been significant interest in deducing the form of the rate laws for chemical reactions occurring in the intracellular environment. This environment is typically characterized by low-dimensionality and a high macromolecular content; this leads to a spatial heterogeneity not typical of the well stirred in vitro environments. For this reason, the classical law of mass action has been presumed to be invalid for modeling intracellular reactions. Using lattice-gas automata models, it has recently been postulated [H. Berry, Monte Carlo simulations of enzyme reactions in two dimensions: Fractal kinetics and spatial segregation, Biophys. J. 83 (2002) 1891-1901; S. Schnell, T.E. Turner, Reaction kinetics in intracellular environments with macromolecular crowding: simulations and rate laws, Prog. Biophys. Mol. Biol. 85 (2004) 235-260] that the reaction kinetics is fractal-like. In this article we systematically investigate for the first time how the rate laws describing intracellular reactions vary as a function of: the geometry and size of the intracellular surface on which the reactions occur, the mobility of the macromolecules responsible for the crowding effects, the initial reactant concentrations and the probability of reaction between two reactant molecules. We also compare the rate laws valid in heterogeneous environments in which there is an underlying spatial lattice, for example crystalline alloys, with the rate laws valid in heterogeneous environments where there is no such natural lattice, for example in intracellular environments. Our simulations indicate that: (i) in intracellular environments both fractal kinetics and mass action can be valid, the major determinant being the probability of reaction, (ii) the geometry and size of the intracellular surface on which reactions are occurring does not significantly affect the rate law, (iii) there are considerable differences between the rate laws valid in heterogeneous non-living structures such as crystals and those valid in intracellular environments. Deviations from mass action are less pronounced in intracellular environments than in a crystalline material of similar heterogeneity.

Microbiology of flooded rice paddies.

Flooded rice paddies are one of the major biogenic sources of atmospheric methane. Apart from this contribution to the 'greenhouse' effect, rice paddy soil represents a suitable model system to study fundamental aspects of microbial ecology, such as diversity, structure, and dynamics of microbial communities as well as structure-function relationships between microbial groups. Flooded rice paddy soil can be considered as a system with three compartments (oxic surface soil, anoxic bulk soil, and rhizosphere) characterized by different physio-chemical conditions. After flooding, oxygen is rapidly depleted in the bulk soil. Anaerobic microorganisms, such as fermentative bacteria and methanogenic archaea, predominate within the microbial community, and thus methane is the final product of anaerobic degradation of organic matter. In the surface soil and the rhizosphere well-defined microscale chemical gradients can be measured. The oxygen profile seems to govern gradients of other electron acceptors (e.g., nitrate, iron(III), and sulfate) and reduced compounds (e.g., ammonium, iron(II), and sulfide). These gradients provide information about the activity and spatial distribution of functional groups of microorganisms. This review presents the current knowledge about the highly complex microbiology of flooded rice paddies. In Section 2 we describe the predominant microbial groups and their function with particular regard to bacterial populations utilizing polysaccharides and simple sugars, and to the methanogenic archaea. Section 3 describes the spatial and temporal development of microscale chemical gradients measured in experimentally defined model systems, including gradients of oxygen and dissolved and solid-phase iron(III) and iron(II). In Section 4, the results of measurements of microscale gradients of oxygen, pH, nitrate-nitrite, and methane in natural rice fields and natural rice soil cores taken to the laboratory will be presented. Finally, perspectives of future research are discussed (Section 5).

[Basic psychosomatic care in ophthalmology. Relevance, training and case examples]. / Psychosomatische Grundversorgung in der Augenheilkunde. Relevanz, Ausbildung, Fallbeispiele.

BACKGROUND: The incidence of psychosomatic disorders and their impact on society are increasing. Many patients suffer from psychosomatic symptoms. Medical studies and most notably medical training for ophthalmologists do not sufficiently cover these topics and do not adequately prepare doctors for dealing with patients suffering from psychosomatic disorders. OBJECTIVE: Training in basic psychosomatic care can be absolved by all physicians irrespective of specialization. The structure, benefits and importance of this professional training are explained. MATERIAL AND METHODS: The curriculum of the German Medical Association forms the basis of training in basic psychosomatic care. The personal experiences of the authors after completing the training as well as case studies are presented. RESULTS AND CONCLUSION: Training in basic psychosomatic care conveys practical skills for dealing with patients with psychosomatic symptoms, which are often not acquired during medical training for ophthalmologists, where technical procedures predominate. Thus the professional ability is broadened with an immediate positive effect not only on the physician-patient relationship but also on the professional and private environment. Training in basic psychosomatic care should be obligatory in the specialist training of ophthalmologists.

Reaction kinetics in intracellular environments with macromolecular crowding: simulations and rate laws.

We review recent evidence illustrating the fundamental difference between cytoplasmic and test tube biochemical kinetics and thermodynamics, and showing the breakdown of the law of mass action and power-law approximation in in vivo conditions. Simulations of biochemical reactions in non-homogeneous media show that as a result of anomalous diffusion and mixing of the biochemical species, reactions follow a fractal-like kinetics. Consequently, the conventional equations for biochemical pathways fail to describe the reactions in in vivo conditions. We present a modification to fractal-like kinetics following the Zipf-Mandelbrot distribution which will enable the modelling and analysis of biochemical reactions occurring in crowded intracellular environments.

Mathematical and computational techniques to deduce complex biochemical reaction mechanisms.

Time series data can now be routinely collected for biochemical reaction pathways, and recently, several methods have been proposed to infer reaction mechanisms for metabolic pathways and networks. In this paper we provide a survey of mathematical techniques for determining reaction mechanisms for time series data on the concentration or abundance of different reacting components, with little prior information about the pathways involved.

Effect of semen thaw method on conception rate in four large commercial dairy heifer herds.

Semen processed with procedures intended to permit a flexible thaw method is used to breed millions of cows yearly. One method of thawing straws, the "pocket thaw" is used extensively with semen prepared with these procedures. Published field data is lacking for thaw method comparisons with semen processed to permit flexible-thawing. The objective of the present study was to measure the effect of semen thaw method (warm-water or pocket thaw) over all seasons and its interaction with herds, inseminators, straw package size, and sperm number on conception rate in commercial dairy heifer herds using semen processed with procedures historically optimized for success with flexible-thawing. Professional inseminators performed 11,215 services over a 16-month period in four large herds, achieving a 67.6% conception rate. Thaw method was alternated weekly. Thaw effect on conception status, determined by 70 days non-return rate, was estimated by a generalized linear mixed model. Neither thaw method nor number of sperm per straw significantly affected probability of conception (P=0.658 and 0.769, respectively). No interactions of thaw method with herd, sperm number, season, straw size, and straw size by season were detected (P=0.297, 0.526, 0.365, 0.723, and 0.824, respectively). Bull, herd, inseminator within herd, year, season, and straw size affected conception rate (P=0.002, 0.000, 0.000, 0.000, 0.000, and 0.014, respectively). In conclusion, for semen processed with procedures that permit flexible-thawing, thaw method (pocket thaw versus warm-water thaw) did not affect conception rate under commercial conditions and with routine semen handling methods.