Assessment of Novel Semen Evaluation Technologies and Breed Comparisons in Yearling Beef Bulls

Objective: The objectives were 1) to evaluate the iSperm, when conducting breeding soundness exams (BSE) on bulls by comparing sperm motility to a technician’s assessment and 2) to evaluate correlations between sperm response to reactive oxygen species (ROS) and functional sperm measurements. Study Description: Ejaculates were collected via electroejaculation from yearling bulls as part of a BSE. All BSE were conducted by one veterinarian and ejaculates were evaluated by a single technician. Additional sperm motility analysis was conducted with the iSperm analyzer. Ejaculates meeting minimum thresholds for passing a BSE were diluted and sent overnight for flow cytometry evaluation. Data were analyzed using Pearson’s correlation coefficients in SAS. Results: Both gross and progressive motilities were significantly (r = 0.30; 0.38; P \u3c 0.001) correlated to the technician’s assessment of progressive motility. Percentage of live spermatozoa with positive ROS status was correlated (r = 0.53; P \u3c 0.001) with percentage progressive motility. Percentage of live spermatozoa with negative ROS status was moderately correlated with percentage spermatozoa exhibiting secondary abnormalities (r = 0.33; P = 0.02). Percentage live spermatozoa that had disrupted acrosomes was strongly correlated with percentage live spermatozoa with negative ROS (r = 0.66; P \u3c 0.001) and moderately negatively correlated with percentage live spermatozoa with positive ROS (r = -0.31; P = 0.04). Percentage of live spermatozoa with positive ROS status was correlated (r = 0.58; P \u3c 0.001) with percentage of spermatozoa with active mitochondrial membranes. Percentage of live spermatozoa with positive ROS status was strongly correlated (r = 0.92; P \u3c 0.001) with percentage of live spermatozoa with intact acrosomes. The Bottom Line: The iSperm can be used to produce semen assessments similar to those of a trained technician and may offer a useful tool for producers to perform on-farm semen analysis. Sperm health and function continue to be related to negative ROS status

Operando and High-throughput multicscale-tomography

We report about multiscale tomography with high throughput at the Diamond beamline I13L. The beamline has the purpose of multi-scale and operando imaging and consists of two independent branchlines operating in real and reciprocal space. The imaging branch -called Diamond-Manchester branchline- hosts micro-tomography, grating interferometry and a full-field microscope. For rapid recording a broad spectrum of the undulator radiation is used either with band-passing the light with a combination of a filter and a deflecting mirror or using a multilayer monochromator. For all the methods similar recording times can be achieved, with typical scanning times of some minutes and covering the resolution range from microns to the 100nm range. Most recently a robot arm has been installed to increase the throughput to 300 samples per day. The system is now implemented for user operation in remote operation mode for the micro-tomography setup and can be expanded to the two other experiments. The instrumental capabilities are applied on various topics such as the study of biodiversity of insects or the structural variations of electrode materials in batteries. Fast recording with dedicated sample environments (not using the sample changing robot) enables operando studies in many areas, the charging/discharging cycles on batteries, the degradation of teeth enamel under various conditions or loading brine sandstone mixtures with CO2, to name some examples. For imaging with highest spatial resolution we managed to improve significantly the recording speed of ptycho-tomography, which is now in the order of hours and will be reduced further. We demonstrated in the past 2-D recording with 10kHz and expand the instrumental capability with specific hardware dependent triggering and scanning schemes. We expand the research program for multi-scale imaging across both branchlines (imaging and coherence branchlines) with first studies such as batteries, brain research, concrete

Femtosecond multimodal imaging with a laser-driven X-ray source

Laser-plasma accelerators are compact linear accelerators based on the interaction of high-power lasers with plasma to form accelerating structures up to 1000 times smaller than standard radiofrequency cavities, and they come with an embedded X-ray source, namely betatron source, with unique properties: small source size and femtosecond pulse duration. A still unexplored possibility to exploit the betatron source comes from combining it with imaging methods able to encode multiple information like transmission and phase into a single-shot acquisition approach. In this work, we combine edge illumination-beam tracking (EI-BT) with a betatron X-ray source and present the demonstration of multimodal imaging (transmission, refraction, and scattering) with a compact light source down to the femtosecond timescale. The advantage of EI-BT is that it allows multimodal X-ray imaging technique, granting access to transmission, refraction and scattering signals from standard low-coherence laboratory X-ray sources in a single shot

Human factors and missed solutions to Enigma design weaknesses

The German World War II Enigma suffered from design weaknesses that facilitated its large-scale decryption by the British throughout the war. The author shows that the main technical weaknesses (self-coding and reciprocal coding) could have been avoided using simple contemporary technology, and therefore the true cause of the weaknesses is not technological but must be sought elsewhere. Specifically, human factors issues resulted in the persistent failure to seek out more effective designs. Similar limitations seem to beset the literature on the period, which misunderstands the Enigma weaknesses and therefore inhibits broader thinking about design or realising the critical role of human factors engineering in cryptography

Preparation of Group I Introns for Biochemical Studies and Crystallization Assays by Native Affinity Purification

The study of functional RNAs of various sizes and structures requires efficient methods for their synthesis and purification. Here, 23 group I intron variants ranging in length from 246 to 341 nucleotides—some containing exons—were subjected to a native purification technique previously applied only to shorter RNAs (<160 nucleotides). For the RNAs containing both exons, we adjusted the original purification protocol to allow for purification of radiolabeled molecules. The resulting RNAs were used in folding assays on native gel electrophoresis and in self-splicing assays. The intron-only RNAs were subjected to the regular native purification scheme, assayed for folding and employed in crystallization screens. All RNAs that contained a 3′ overhang of one nucleotide were efficiently cleaved off from the support and were at least 90% pure after the non-denaturing purification. A representative subset of these RNAs was shown to be folded and self-splicing after purification. Additionally, crystals were grown for a 286 nucleotide long variant of the Clostridium botulinum intron. These results demonstrate the suitability of the native affinity purification method for the preparation of group I introns. We hope these findings will stimulate a broader application of this strategy to the preparation of other large RNA molecules

Diclofenac Prolongs Repolarization in Ventricular Muscle with Impaired Repolarization Reserve

Background: The aim of the present work was to characterize the electrophysiological effects of the non-steroidal anti- inflammatory drug diclofenac and to study the possible proarrhythmic potency of the drug in ventricular muscle. Methods: Ion currents were recorded using voltage clamp technique in canine single ventricular cells and action potentials were obtained from canine ventricular preparations using microelectrodes. The proarrhythmic potency of the drug was investigated in an anaesthetized rabbit proarrhythmia model. Results: Action potentials were slightly lengthened in ventricular muscle but were shortened in Purkinje fibers by diclofenac (20 mM). The maximum upstroke velocity was decreased in both preparations. Larger repolarization prolongation was observed when repolarization reserve was impaired by previous BaCl 2 application. Diclofenac (3 mg/kg) did not prolong while dofetilide (25 mg/kg) significantly lengthened the QT c interval in anaesthetized rabbits. The addition of diclofenac following reduction of repolarization reserve by dofetilide further prolonged QT c . Diclofenac alone did not induce Torsades de Pointes ventricular tachycardia (TdP) while TdP incidence following dofetilide was 20%. However, the combination of diclofenac and dofetilide significantly increased TdP incidence (62%). In single ventricular cells diclofenac (30 mM) decreased the amplitude of rapid (I Kr ) and slow (I Ks ) delayed rectifier currents thereby attenuating repolarization reserve. L-type calcium current (I Ca ) was slightly diminished, but the transient outward (I to ) and inward rectifier (I K1 ) potassium currents were not influenced. Conclusions: Diclofenac at therapeutic concentrations and even at high dose does not prolong repolarization markedly and does not increase the risk of arrhythmia in normal heart. However, high dose diclofenac treatment may lengthen repolarization and enhance proarrhythmic risk in hearts with reduced repolarization reserve