Bacterial killing activity and lysozymes: A stable defence mechanism in stallion seminal plasma?

During insemination, bacterial contamination of the ejaculate can lead to reduced sperm quality and transmission of pathogens to the female, thus should be avoided. The semen of a variety of animal taxa possess antimicrobial properties against a wide range of bacterial species through antimicrobial molecules, such as lysozyme, but their variance and the factors influencing it are unknown for most species. In this study, the antibacterial defence (bacterial killing activity (BKA) against Escherichia (E.) coli and Staphylococcus (S.) aureus as well as lysozyme concentration) was studied in seminal fluid from two consecutive ejaculates of 18 stallions. All ejaculates showed BKA against the tested bacteria, which correlated between the two consecutive ejaculates (r_S = 0.526, p = .025 for E. coli and r_S = 0.656, p = .003 for S. aureus) and appeared to be stable over the tested period. The lysozyme concentration (LC) showed no significant correlation between the consecutive ejaculates (r_S = 0.161, p = .681). However, LC had a positive correlation to the ratio of apoptotic spermatozoa within the ejaculates (r_S = 0.426, p = .019). In contrast to other livestock (e.g., boar, bull), the BKA in stallion semen did not correlate significantly with the age of the animal nor sperm quality characteristics.     

The effect of low dose rocuronium on globe position, muscle relaxation and ventilation in dogs: a clinical study

OBJECTIVE: The purpose of this study was to evaluate globe position, muscle relaxation and changes in ventilatory parameters after intravenous administration of 0.1 mg/kg rocuronium. STUDY DESIGN: Prospective clinical study. ANIMAL STUDIED: Sixteen dogs of different breeds, with a body weight of 22.1 +/- 13 kg and age of 5.6 +/- 2.8 years (mean +/- SD), were anesthetized for a short ophthalmic examination requiring central position of the globe. PROCEDURES: All dogs were premedicated with 0.005 mg/kg medetomidine and 0.1 mg/kg methadone IV. Anesthesia was induced with propofol to effect and maintained with 10 mg/kg/h propofol by continuous rate infusion. Following endotracheal intubation all dogs breathed 100% oxygen via an anesthetic circle system. Neuromuscular function was assessed with an acceleromyograph (TOF-Guard, Organon Teknika NV, Turnhout, Belgium) and by stimulation of the nervus peroneus superficialis. The ventilation parameters were measured using spirometry and capnography. After baseline measurements 0.1 mg/kg rocuronium was administered IV. Minute volume (MV), tidal volume (Vt), respiratory rate (RR), end expiratory carbon dioxide concentration (PE'CO(2)) and maximal depression of the response of the first twitch (T1) of train-of-four (TOF) stimulation and train-of-four ratio (TOFR) was measured. The change in the position of the globe was recorded. RESULTS: T1 decreased to 61 +/- 18% and the TOF ratio to 45 +/- 21% of baseline values. Both parameters returned to baseline after 9 min. There was no significant reduction in MV, TV and RR and no increase in PE'CO(2). The globe rotated to a central position of 45 +/- 7.7 s after administration of rocuronium and remained there for 23 +/- 10.8 min in all dogs. CONCLUSION: Rocuronium administered intravenously at a dose of 0.1 mg/kg to dogs causes a central position of the globe but minimal impairment of ventilation parameters.     

Reversal of neuromuscular block in companion animals

ObjectiveTo review the evidence regarding the reversal of neuromuscular block (NMB) in companion animals with emphasis on the development and use of newer agents.Database usedData sources include scientific reviews and original research publications in both human and veterinary literature using Pubmed and Scopus as search data bases. Unpublished and locally published data on reversal of NMB are presented.ConclusionsResidual NMB has been shown to increase morbidity and mortality in humans and needs to be avoided. It can be detected only by adequate neuromuscular monitoring. The proper use of reversal agents avoids residual NMB and recurarization should not occur. Anticholinesterase inhibitors, such as edrophonium and neostigmine have been used to reverse NMB when the need for this has been established. Reversal is influenced by several factors and a number of undesirable side- effects of these drugs have been reported. Sugammadex, a γ-cyclodextrin, which was designed specifically to encapsulate rocuronium, is more rapid in its actions, has fewer side effects and can reverse profound NMB induced by aminosteroidal muscle relaxants.     

In search for the ecological and toxicological relevance of sediment re-mobilisation and transport during flood events

In response to increasing concerns about the potential toxicological impacts of (extreme) flood events, scientists from several disciplines have joined to form the interdisciplinary research project named FLOODSEARCH. FLOODSEARCH is one of the recent Pathfinder Projects supported by the German Excellence Initiative via the Exploratory Research Space at RWTH Aachen (ERS). FLOODSEARCH aims to combine methodologies of hydraulic engineering and ecotoxicology in a new interdisciplinary approach to assess the risks associated with the re-mobilisation of particulate bound contaminants often observed after severe flood events. Impacts of extreme flood events and aspects of re-mobilisation of sediment-bound toxic compounds will be characterised and evaluated in controlled experiments fusing flood simulation technologies with biological effects assessment. The overall goal is to establish a novel and more realistic approach towards flood event testing that can be applied to a number of different questions and species. Specifically, model aquatic species such as rainbow trout (Onchorhynchus mykiss) will be exposed to particle-bound contaminants in flood-like conditions in a specifically designed annular flume that permits monitoring of both physical/chemical and biological parameter. Ultimately, this approach will assist to further our understanding of the potential biological risks associated with increasingly frequent extreme flood events, e.g., as a consequence of climate change, by bridging the gap between the physical (re-)mobilisation of contaminants and resulting toxicological impacts on aquatic organisms. Thus, it is the objective of the project to derive relationships between the hydrodynamic parameters such as velocities and turbulences, the parameters associated to sediment transport such as sediment concentration and grain sizes and the biological parameters.     

Metabolism and persistence of atrazine in several field soils with different atrazine application histories

To assess the potential occurrence of accelerated herbicide degradation in soils, the mineralization and persistence of (14)C-labeled and nonlabeled atrazine was evaluated over 3 months in two soils from Belgium (BS, atrazine-treated 1973-2008; BC, nontreated) and two soils from Germany (CK, atrazine-treated 1986-1989; CM, nontreated). Prior to the experiment, accelerated solvent extraction of bulk field soils revealed atrazine (8.3 and 15.2 μg kg(-1)) in BS and CK soils and a number of metabolites directly after field sampling, even in BC and CM soils without previous atrazine treatment, by means of LC-MS/MS analyses. For atrazine degradation studies, all soils were incubated under different moisture conditions (50% maximum soil water-holding capacity (WHC(max))/slurried conditions). At the end of the incubation, the (14)C-atrazine mineralization was high in BS soil (81 and 83%) and also unexpectedly high in BC soil (40 and 81%), at 50% WHC(max) and slurried conditions, respectively. In CK soil, the (14)C-atrazine mineralization was higher (10 and 6%) than in CM soil (4.7 and 2.7%), but was not stimulated by slurried conditions. The results revealed that atrazine application history dramatically influences its degradation and mineralization. For the incubation period, the amount of extractable atrazine, composed of residues from freshly applied atrazine and residues from former field applications, remained significantly greater (statistical significance = 99.5 and 99.95%) for BS and CK soils, respectively, than the amount of extractable atrazine in the bulk field soils. This suggests that (i) mostly freshly applied atrazine is accessible for a complex microbial community, (ii) the applied atrazine is not completely mineralized and remains extractable even in adapted soils, and (iii) the microbial atrazine-mineralizing capacity strongly depends on atrazine application history and appears to be conserved on long time scales after the last application.     

A comparison of cardiorespiratory variables during isoflurane-fentanyl and propofol-fentanyl anaesthesia for surgery in injured cats

OBJECTIVE: To compare haemodynamic and respiratory variables during isoflurane-fentanyl (IF) and propofol-fentanyl (PF) anaesthesia for surgery in injured cats. STUDY DESIGN: Prospective, randomized, controlled clinical study. ANIMALS: Thirty-three client-owned injured cats undergoing orthopaedic surgery. MATERIALS AND METHODS: Pre-anaesthetic medication was intravenous midazolam 1 mg kg(-1), butorphanol 0.4 mg kg(-1) and ketamine 2 mg kg(-1). Anaesthesia was induced with propofol (P) and maintained with either: (a) a continuous rate infusion (CRI) of fentanyl (F) 0.02 mg kg(-1) hour(-1) and isoflurane (initial end-tidal concentration of 1%), (b) a fentanyl CRI (dose as before) and sevoflurane (initial end-tidal concentration of 2%) or (c) a CRI of propofol (12 mg kg(-1) hour(-1)). All three techniques were given to effect until surgical anaesthesia was achieved. Heart rate and rhythm (ECG), mean arterial blood pressure, respiratory rate, tidal volume and end-tidal CO(2) concentration were recorded. Venous blood gas analysis was performed before and after sedation, and at the end of anaesthesia. Blood chemistry and blood cell counts were assessed before, at the end of, and 24 hours after anaesthesia. The variables recorded from cats anaesthetized with IF and PF were compared. RESULTS: Mean end-expiratory isoflurane concentration was 1.19 +/- 0.19%. The propofol infusion rate was 11.4 +/- 0.8 mg kg(-1) hour(-1). No significant differences between the two groups in heart rate were identified; no cardiac dysrhythmias were recorded. Mean arterial blood pressure was significantly lower in IF cats during skin incision (p = 0.01), during surgery without intense surgical stimulation (p < 0.01) and during surgery with intense surgical stimulation (p = 0.01). Nine of 11 cats in the IF group were markedly hypotensive (34-49 mmHg) while seven of 11 cats in group PF were mildly hypotensive (49-59 mmHg). One of 11 cats in group IF and nine of 11 cats in group PF required intermittent positive pressure ventilation (IPPV) to maintain end-tidal CO(2) levels below 6.66 kPa (50 mmHg). CONCLUSION AND CLINICAL RELEVANCE: Despite the necessity to ventilate the lungs of cats in the PF group, arterial blood pressure was better maintained. Propofol-fentanyl anaesthesia is better for surgery in injured cats providing the means to impose IPPV are available.     

Heat-induced changes in myofibrillar protein structures and myowater of two pork qualities. A combined FT-IR spectroscopy and low-field NMR relaxometry study

Low-field NMR T(2) and Fourier transform infrared (FT-IR) measurements were performed on meat samples of two qualities (normal and high ultimate pH) during cooking from 28 degrees C to 81 degrees C. Pronounced changes in both T(2) relaxation data and FT-IR spectroscopic data were observed during cooking, revealing severe changes in the water properties and structural organization of proteins. The FT-IR data revealed major changes in bands in the amide I region (1700-1600 cm(-)(1)), and a tentative assignment of these is discussed. Distributed NMR T(2) relaxation data and FT-IR spectra were compared by partial least-squares regression. This revealed a correlation between the FT-IR peaks reflecting beta-sheet and alpha-helix structures and the NMR relaxation populations reflecting hydration water (T(2B) approximately 0-10 ms), myofibrillar water (T(21) approximately 35-50 ms), and also expelled "bulk" water (T(2) relaxation times >1000 ms). Accordingly, the present study demonstrates that definite structural changes in proteins during cooking of meat are associated with simultaneous alterations in the chemical-physical properties of the water within the meat.     

Influence of aging and salting on protein secondary structures and water distribution in uncooked and cooked pork. A combined FT-IR microspectroscopy and 1H NMR relaxometry study

Fourier transform infrared (FT-IR) microspectroscopy and low-field (LF) proton NMR transverse relaxation measurements were used to study the changes in protein secondary structure and water distribution as a consequence of aging (1 day and 14 days) followed by salting (3%, 6%, and 9% NaCl) and cooking (65 degrees C). An enhanced water uptake and increased proton NMR relaxation times after salting were observed in aged meat (14 days) compared with nonaged meat (1 day). FT-IR bands revealed that salting induced an increase in native beta-sheet structure while aging triggered an increase in native alpha-helical structure before cooking, which could explain the effects of aging and salting on water distribution and water uptake. Moreover, the decrease in T2 relaxation times and loss of water upon cooking were attributed to an increase in aggregated beta-sheet structures and a simultaneous decrease in native protein structures. Finally, aging increased the cooking loss and subsequently decreased the final yield, which corresponded to a further decrease in T2 relaxation times in aged meat upon cooking. However, salting weakened the effect of aging on the final yield, which is consistent with the increased T2 relaxation times upon salting for aged meat after cooking and the weaker effect of aging on protein secondary structural changes for samples treated with high salt concentration. The present study reveals that changes in water distribution during aging, salting, and cooking are not only due to the accepted causal connection, i.e., proteolytic degradation of myofibrillar structures, change in electrostatic repulsion, and dissolution and denaturation of proteins, but also dynamic changes in specific protein secondary structures.     

Structural basis for prereceptor modulation of plant hormones by GH3 proteins

Acyl acid amido synthetases of the GH3 family act as critical prereceptor modulators of plant hormone action; however, the molecular basis for their hormone selectivity is unclear. Here, we report the crystal structures of benzoate-specific Arabidopsis thaliana AtGH3.12/PBS3 and jasmonic acid-specific AtGH3.11/JAR1. These structures, combined with biochemical analysis, define features for the conjugation of amino acids to diverse acyl acid substrates and highlight the importance of conformational changes in the carboxyl-terminal domain for catalysis. We also identify residues forming the acyl acid binding site across the GH3 family and residues critical for amino acid recognition. Our results demonstrate how a highly adaptable three-dimensional scaffold is used for the evolution of promiscuous activity across an enzyme family for modulation of plant signaling molecules.