Prevalence and Antibiotic Susceptibility of Bacterial Isolates From Dogs With Ulcerative Keratitis in Midwestern United States.

The objective of this study was to describe bacterial culture and antibiotic susceptibility results in 476 dogs presenting with suspected bacterial keratitis in Iowa and surrounding Midwestern states, further detailing trends in patient characteristics, seasonality, and antimicrobial resistance. Corneal swabs yielded 465 bacterial isolates and 220 cultures (46.2%) with no apparent growth (0-5 isolates per culture). The most frequent bacterial genera were Staphylococcus (32.3%), Streptococcus (19.1%), and Pseudomonas (12.5%), while the most common bacterial species were Staphylococcus pseudintermedius (26.7%), Streptococcus canis (12%), and Pseudomonas aeruginosa (7.5%). Compared to mixed-breed dogs, canine breeds most likely to be examined for ulcerative keratitis included Boston terrier, Cavalier King Charles spaniel, miniature pinscher, pug, rat terrier, Saint Bernard, shih tzu, and silky terriers. In summer, the likelihood to yield a negative culture was reduced while the likelihood to culture Pseudomonas species was increased. Bacteria considered multidrug resistant (MDR, resistant to ≥ 3 antibiotic classes) represented 20% of all canine isolates and were most prevalent for Staphylococcus species (33%). An alarming, escalating trend of MDR prevalence was noted between 2016 (5%) and 2020 (34%). Individual ophthalmic preparations (i.e., single antibiotics or commercially available antibiotic combinations) with highest efficacy against all bacterial isolates included chloramphenicol (83%), ceftiofur (79%), amikacin (77%), neomycin-polymyxin B-bacitracin (77%), and gentamicin (74%). Efficacy of systemic antibiotics and combinations of ophthalmic preparations was also evaluated. Based on the present findings, triple antibiotic (Neo-Poly-Bac) is recommended as empirical monotherapy for prophylactic antibiotic therapy in dogs with simple corneal ulcers, while a chloramphenicol-ciprofloxacin combination is empirically recommended for therapeutic management of infected corneal ulcers. Pending culture and susceptibility results, appropriate selection of empiric antibiotic therapy is important to enhance therapeutic outcome and reduce antibacterial resistance in dogs with corneal ulceration.

Rapid paper spray mass spectrometry characterization of uranium and exemplar molecular species.

RATIONALE: The ability to detect and quantify the presence of specific inorganic elements and complexes is essential for environmental monitoring and nuclear safeguards applications. In this work, paper spray ionization mass spectrometry was used for the rapid chemical and isotopic characterization of trace inorganic species collected on cotton swipe substrates. The direct analysis of cotton swipes using this ambient ionization technique led to fast sample analysis that retained original chemical information of the source material with minimal sample preparation. METHODS: Mass spectra were collected with an atmospheric pressure ionization, high-resolution mass spectrometer for solutions containing uranyl acetate, uranyl chloride, uranyl nitrate, and uranyl tri-n-butylphosphate complexes. Gadolinium nitrate was used as an internal standard for the quantitative analysis of uranium. To demonstrate the ability to characterize inorganic contaminants in the presence of uranium, a multi-element inorganic standard containing U, Bi, Pb, Cd, Fe, and Zn was deposited onto cotton substrates and directly analyzed without purification. RESULTS: All elements doped on the cotton substrate were detected with strong signal-to-noise ratios (ca 1000 for UO2 + on multi-element doped swipes) and high integrated intensities (>105 counts) from collection periods of approximately 1 min. Limits of detection were determined to be approximately 94 ng for UO2 + and uranyl acetate through the measurement of ppb level solutions. CONCLUSIONS: The rapid analysis of uranium and other inorganic-containing samples while still retaining original chemical information (e.g. uranyl complexation) was demonstrated. Qualitative detection and speciation were achieved in less than 1 min of analysis. For uranium isotopic quantitation, longer accumulations (>15 min) can be sustained to improve the accuracy of minor 235 U isotopic abundance measurements to approximately 1% error.

A prime-boost concept using a T-cell epitope-driven DNA vaccine followed by a whole virus vaccine effectively protected pigs in the pandemic H1N1 pig challenge model.

Influenza A virus (IAV) vaccines in pigs generally provide homosubtypic protection but fail to prevent heterologous infections. In this pilot study, the efficacy of an intradermal pDNA vaccine composed of conserved SLA class I and class II T cell epitopes (EPITOPE) against a homosubtypic challenge was compared to an intramuscular commercial inactivated whole virus vaccine (INACT) and a heterologous prime boost approach using both vaccines. Thirty-nine IAV-free, 3-week-old pigs were randomly assigned to one of five groups including NEG-CONTROL (unvaccinated, sham-challenged), INACT-INACT-IAV (vaccinated with FluSure XP® at 4 and 7 weeks, pH1N1 challenged), EPITOPE-INACT-IAV (vaccinated with PigMatrix EDV at 4 and FluSure XP® at 7 weeks, pH1N1 challenged), EPITOPE-EPITOPE-IAV (vaccinated with PigMatrix EDV at 4 and 7 weeks, pH1N1 challenged), and a POS-CONTROL group (unvaccinated, pH1N1 challenged). The challenge was done at 9 weeks of age and pigs were necropsied at day post challenge (dpc) 5. At the time of challenge, all INACT-INACT-IAV pigs, and by dpc 5 all EPITOPE-INACT-IAV pigs were IAV seropositive. IFNγ secreting cells, recognizing vaccine epitope-specific peptides and pH1N1 challenge virus were highest in the EPITOPE-INACT-IAV pigs at challenge. Macroscopic lung lesion scores were reduced in all EPITOPE-INACT-IAV pigs while INACT-INACT-IAV pigs exhibited a bimodal distribution of low and high scores akin to naïve challenged animals. No IAV antigen in lung tissues was detected at necropsy in the EPITOPE-INACT-IAV group, which was similar to naïve unchallenged pigs and different from all other challenged groups. Results suggest that the heterologous prime boost approach using an epitope-driven DNA vaccine followed by an inactivated vaccine was effective against a homosubtypic challenge, and further exploration of this vaccine approach as a practical control measure against heterosubtypic IAV infections is warranted.

Somewhere I belong: Long-term increases in adolescents' resilience are predicted by perceived belonging to the in-group.

This study sought to examine the role of belonging in the increases in resilience observed following an adventure education programme (AEP). First, we demonstrate that group belonging makes a significant contribution to the improvement in resilience participants' experienced over the course of the AEP. Second, we demonstrate that this increase in resilience is maintained 9 months following the AEP and that group belonging maintained a significant contribution when controlling for participants' initial resilience level and other psychosocial variables (i.e., centrality of identity and social support). Our findings accord well with recent research on the Social Cure or Social Identity Approach to Health and add to a growing body of work identifying the mechanisms underlying this phenomenon.

Calculation of retention time tolerance windows with absolute confidence from shared liquid chromatographic retention data.

Compound identification by liquid chromatography-mass spectrometry (LC-MS) is a tedious process, mainly because authentic standards must be run on a user's system to be able to confidently reject a potential identity from its retention time and mass spectral properties. Instead, it would be preferable to use shared retention time/index data to narrow down the identity, but shared data cannot be used to reject candidates with an absolute level of confidence because the data are strongly affected by differences between HPLC systems and experimental conditions. However, a technique called "retention projection" was recently shown to account for many of the differences. In this manuscript, we discuss an approach to calculate appropriate retention time tolerance windows for projected retention times, potentially making it possible to exclude candidates with an absolute level of confidence, without needing to have authentic standards of each candidate on hand. In a range of multi-segment gradients and flow rates run among seven different labs, the new approach calculated tolerance windows that were significantly more appropriate for each retention projection than global tolerance windows calculated for retention projections or linear retention indices. Though there were still some small differences between the labs that evidently were not taken into account, the calculated tolerance windows only needed to be relaxed by 50% to make them appropriate for all labs. Even then, 42% of the tolerance windows calculated in this study without standards were narrower than those required by WADA for positive identification, where standards must be run contemporaneously.

"Measure Your Gradient": a new way to measure gradients in high performance liquid chromatography by mass spectrometric or absorbance detection.

The gradient produced by an HPLC is never the same as the one it is programmed to produce, but non-idealities in the gradient can be taken into account if they are measured. Such measurements are routine, yet only one general approach has been described to make them: both HPLC solvents are replaced with water, solvent B is spiked with 0.1% acetone, and the gradient is measured by UV absorbance. Despite the widespread use of this procedure, we found a number of problems and complications with it, mostly stemming from the fact that it measures the gradient under abnormal conditions (e.g. both solvents are water). It is also generally not amenable to MS detection, leaving those with only an MS detector no way to accurately measure their gradients. We describe a new approach called "Measure Your Gradient" that potentially solves these problems. One runs a test mixture containing 20 standards on a standard stationary phase and enters their gradient retention times into open-source software available at www.measureyourgradient.org. The software uses the retention times to back-calculate the gradient that was truly produced by the HPLC. Here we present a preliminary investigation of the new approach. We found that gradients measured this way are comparable to those measured by a more accurate, albeit impractical, version of the conventional approach. The new procedure worked with different gradients, flow rates, column lengths, inner diameters, on two different HPLCs, and with six different batches of the standard stationary phase.

A practical methodology to measure unbiased gas chromatographic retention factor vs. temperature relationships.

Compound identification continues to be a major challenge. Gas chromatography-mass spectrometry (GC-MS) is a primary tool used for this purpose, but the GC retention information it provides is underutilized because existing retention databases are experimentally restrictive and unreliable. A methodology called "retention projection" has the potential to overcome these limitations, but it requires the retention factor (k) vs. T relationship of a compound to calculate its retention time. Direct methods of measuring k vs. T relationships from a series of isothermal runs are tedious and time-consuming. Instead, a series of temperature programs can be used to quickly measure the k vs. T relationships, but they are generally not as accurate when measured this way because they are strongly biased by non-ideal behavior of the GC system in each of the runs. In this work, we overcome that problem by using the retention times of 25 n-alkanes to back-calculate the effective temperature profile and hold-up time vs. T profiles produced in each of the six temperature programs. When the profiles were measured this way and taken into account, the k vs. T relationships measured from each of two different GC-MS instruments were nearly as accurate as the ones measured isothermally, showing less than two-fold more error. Furthermore, temperature-programmed retention times calculated in five other laboratories from the new k vs. T relationships had the same distribution of error as when they were calculated from k vs. T relationships measured isothermally. Free software was developed to make the methodology easy to use. The new methodology potentially provides a relatively fast and easy way to measure unbiased k vs. T relationships.

Oxidative stress is not a major contributor to somatic mitochondrial DNA mutations.

The accumulation of somatic mitochondrial DNA (mtDNA) mutations is implicated in aging and common diseases of the elderly, including cancer and neurodegenerative disease. However, the mechanisms that influence the frequency of somatic mtDNA mutations are poorly understood. To develop a simple invertebrate model system to address this matter, we used the Random Mutation Capture (RMC) assay to characterize the age-dependent frequency and distribution of mtDNA mutations in the fruit fly Drosophila melanogaster. Because oxidative stress is a major suspect in the age-dependent accumulation of somatic mtDNA mutations, we also used the RMC assay to explore the influence of oxidative stress on the somatic mtDNA mutation frequency. We found that many of the features associated with mtDNA mutations in vertebrates are conserved in Drosophila, including a comparable somatic mtDNA mutation frequency (∼10(-5)), an increased frequency of mtDNA mutations with age, and a prevalence of transition mutations. Only a small fraction of the mtDNA mutations detected in young or old animals were G∶C to T∶A transversions, a signature of oxidative damage, and loss-of-function mutations in the mitochondrial superoxide dismutase, Sod2, had no detectable influence on the somatic mtDNA mutation frequency. Moreover, a loss-of-function mutation in Ogg1, which encodes a DNA repair enzyme that removes oxidatively damaged deoxyguanosine residues (8-hydroxy-2'-deoxyguanosine), did not significantly influence the somatic mtDNA mutation frequency of Sod2 mutants. Together, these findings indicate that oxidative stress is not a major cause of somatic mtDNA mutations. Our data instead suggests that somatic mtDNA mutations arise primarily from errors that occur during mtDNA replication. Further studies using Drosophila should aid in the identification of factors that influence the frequency of somatic mtDNA mutations.

Inverse kinetic isotope effect in the excited-state relaxation of a Ru(II)-aquo complex: revealing the impact of hydrogen-bond dynamics on nonradiative decay.

Photophysics of the MLCT excited-state of [Ru(bpy)(tpy)(OH2)](2+) (1) and [Ru(bpy)(tpy)(OD2)](2+) (2) (bpy = 2,2'-bipyridine and tpy = 2,2':6',2″-terpyridine) have been investigated in room-temperature H2O and D2O using ultrafast transient pump-probe spectroscopy. An inverse isotope effect is observed in the ground-state recovery for the two complexes. These data indicate control of excited-state lifetime via a pre-equilibrium between the (3)MLCT state that initiates H-bond dynamics with the solvent and the (3)MC state that serves as the principal pathway for nonradiative decay.

Dynamics of the 3MLCT in Ru(II) terpyridyl complexes probed by ultrafast spectroscopy: evidence of excited-state equilibration and interligand electron transfer.

Ground- and excited-state properties of [Ru(tpy)(2)](2+), [Ru(tpy)(ttpy)](2+), and [Ru(ttpy)(2)](2+) (where tpy = 2,2':6',2″-terpyridine and ttpy = 4'-(4-methylphenyl)-2,2':6',2″-terpyridine) in room temperature acetonitrile have been investigated using linear absorption, electrochemical, and ultrafast transient pump-probe techniques. Spectroelectrochemistry was used to assign features observed in the transient spectra while single wavelength kinetics collected at a variety of probe wavelengths were used to monitor temporal evolution of the MLCT excited state. From these data, the excited-state lifetime of each complex was recovered and the rate limiting decay step was identified. In the bis-heteroleptic complex [Ru(tpy)(ttpy)](2+), photoexcitation to the (1)MLCT manifold generates both tpy-localized and ttpy-localized excited states. Accordingly, interligand electron transfer (ILET) from tpy-localized to the ttpy-localized (3)MLCT excited states is observable and the time scale has been measured to be 3 ps. For the homoleptic complex [Ru(tpy)(2)](2+), evidence for equilibration of the (3)MLCT excited-state population with the (3)MC has been observed and the time scale is reported at 2 ps.

Exploiting conformational dynamics to facilitate formation and trapping of electron-transfer photoproducts in metal complexes.

Three new photoinduced electron donor-acceptor (D-A) systems are reported which juxtapose a Ru(II) excited-state donor with a bipyridinium acceptor via a conformationally active asymmetric aryl-substituted bipyridine ligand participating in the bridge between D and A. Across the series of complexes 1-3, steric bulk is sequentially added to tune the inter-ring dihedral angle theta between the bipyridine and the aryl substituent. Driving forces for photoinduced electron transfer (DeltaG(ET)) and back electron transfer (DeltaG(BET)) are reported based on electrochemical measurements of 1-3 as well as Franck-Condon analysis of emission spectra collected for three new donor model complexes 1'-3'. These preserve the substitution patterns on the aryl substituent in their respective D-A complexes but remove the bipyridinium acceptor. Both DeltaG(ET) and DeltaG(BET) are invariant to within 0.02 eV across the series. Upon visible photoexcitation of each of the D-A systems with approximately 100 fs laser pulses at 500 +/- 10 nm, an electron-transfer (ET) photoproduct is observed to form with a time constant of tau(ET) = 29 ps (1), 37 ps (2), and 57 ps (3). That ET remains relatively rapid throughout this series, even as steric bulk significantly increases the inter-ring dihedral angle theta, is attributed to the effects of ligand-based torsional dynamics driven by intraligand electron delocalization in the D*-A excited state manifold prior to ET. The lifetimes of the charge-separated states (tau(BET)) are also reported with tau(BET) = 98 ps (1), 217 ps (2), and 789 ps (3), representing a more than 8-fold increase across the series. This is attributed to reverse conformational dynamics in D(+)-A(-) driven by steric repulsions, which serves to minimize electronic coupling to the ground state. Steric control of ligand geometry and the range over which theta changes during conformational dynamics provides a new strategy to facilitate the formation and storage of charge-separated excited states.

Template Registration with Missing Parts: Application to the Segmentation of M. Tuberculosis Infected Lungs.

Many techniques have been proposed to segment organs from images, however the segmentation of diseased organs remains challenging and frequently requires lots of user interaction. The challenge consists of segmenting an organ while its appearance and its shape vary due to the presence of the disease in addition to individual variations. We propose a template registration technique that can be used to recover the complete segmentation of a diseased organ from a partial segmentation. The usual template registration method is modified in such a way that it is robust to missing parts. The proposed method is used to segment Mycobacterium tuberculosis infected lungs in CT images of experimentally infected mice. Using synthetic data, we evaluate and compare the performance of the proposed algorithm with the usual sum of squared difference cost function.