Detection of singlet oxygen in blood serum samples of clinically healthy lambs and lambs suffering from alveld disease

Alveld is a disease in lambs of domestic sheep (Ovis aries L.), characterized by a combination of photosensitivity and liver damage. Generation of singlet oxygen play a major role in phototoxicity reactions. The compound phylloerythrin (phytoporphyrin) is so far assumed to be the main photodynamic agent in hepatogenous photosensitivity diseases in sheep. Phylloerythrin is a potent photosensitizer and an efficient source of singlet oxygen. The compound accumulates in the peripheral circualtion upon liver damage. Liver dysfunction is also likely to cause an increase in the blood level of bilirubin. Formation of singlet oxygen by bilirubin is reported. In the present work the photosensitizing potential of serum has been measured and related to the bilirubin- and phylloerythrin levels in lambs suffering from alveld and in clinically healthy controls. The singlet oxygen level of the serum was taken as a measure of the photosensitizing potential. The observed singlet oxygen values in serum from alveld lambs were significantly higher than the corresponding values observed in clinically healthy control lambs. This indicates that the serum of the alveld lambs contains an elevated concentration of photosensitizer. The singlet oxygen level was not correlated to the concentration of bilirubin or phylloerythrin. The results indicate that the photosensitizing mechanism is quite complex and may involve other sensitizer(s) than phylloerythrin.     

Premature death, risk factors, and life patterns in dogs with epilepsy

BACKGROUND: Epilepsy in dogs is often difficult to medically control, resulting in premature death of dogs with epilepsy. However, the risks of premature death are not known. HYPOTHESIS: Dogs with epilepsy have an increased risk of premature death as compared to a general population of dogs. ANIMALS: Sixty-three dogs diagnosed with epilepsy between 1993 and 1996 were included in this study. METHODS: A prospective longitudinal study of the population was performed from the diagnosis of epilepsy until the time of euthanasia, death, or a maximum of 12 years to investigate mortality and risk factors. Information about sex, onset, type, frequency, and control of seizures, remission of epilepsy, death, cause of death, and owner's perspective was collected and analyzed. RESULTS: The median age at death of dogs was 7.0 years. The life span of dogs in which euthanasia or death was directly caused by their epileptic condition was significantly shorter as compared with epileptic dogs that were euthanized because of other causes (P = .001). The median number of years that a dog lived with epilepsy was 2.3 years. Females lived longer with epilepsy than males (P = .036). Seizure type (primary generalized versus focal seizures) was not significantly associated with survival time. The remission rate of epilepsy (spontaneous remission and remission with treatment) was 15%. CONCLUSION AND CLINICAL IMPORTANCE: The diagnosis of epilepsy implies an increased risk of premature death. The prognosis for dogs with epilepsy is dependent on a combination of veterinary expertise, therapeutic success, and the owner's motivation.     

SOS response induction by beta-lactams and bacterial defense against antibiotic lethality

The SOS response aids bacterial propagation by inhibiting cell division during repair of DNA damage. We report that inactivation of the ftsI gene product, penicillin binding protein 3, by either beta-lactam antibiotics or genetic mutation induces SOS in Escherichia coli through the DpiBA two-component signal transduction system. This event, which requires the SOS-promoting recA and lexA genes as well as dpiA, transiently halts bacterial cell division, enabling survival to otherwise lethal antibiotic exposure. Our findings reveal defective cell wall synthesis as an unexpected initiator of the bacterial SOS response, indicate that beta-lactam antibiotics are extracellular stimuli of this response, and demonstrate a novel mechanism for mitigation of antimicrobial lethality.     

Cloning by cuttings in Nordmann fir,Abies nordmanniana: hormonal characteristics in relation crown position,rooting competence,and orthotropism as ramets

New Forests - Conifer tree species belonging to Abies are notoriously difficult to propagate vegetatively due to aging of material, slow rooting, and persistent plagiotropism in rooted cuttings. We...     

Combined high-field 13C CP MAS NMR and low-field NMR relaxation measurements on post mortem porcine muscles

Changes in postmortem muscle characteristics are investigated in muscles from eight pigs exposed to different combinations of preslaughter stress (exercise on treadmill) and stunning method (CO(2) vs electrical stunning). Solid-state (13)C cross-polarization (CP) magic-angle spinning (MAS) NMR experiments are carried out on a total of 16 rapidly frozen M. longissimus muscle biopsies taken in vivo the day before slaughter and at 45 min postmortem. Simultaneously, low-field NMR T(2) relaxation time measurements are carried out on samples from M. longissimus. Glycogen and lactate are estimated from the (13)C CP MAS spectra, and correlations of r = 0.89 and r = 0.70, respectively, to subsequent biochemical determinations using partial least squares regression (PLSR) are established. Moreover, PLSR reveals that, besides the 72 ppm signal (carbons in glycogen), a signal around 38 ppm, which increases concomitantly with lactate, is also significantly correlated to changes in glycogen/lactate. With the assumption that the 38 ppm signal reflects CH(2) in phosphocreatine/creatine, altered mobility of creatine as a result of dephosphorylation is indicated. Finally, PLSR on the 45 min (13)C CP MAS spectra also reveals correlation (r = 0.54) to the slowest relaxing T(2) population (50 min postmortem), known to reflect extra-myofibrillar water. Subsequently, evaluation of the loading plot in the PLSR analysis reveals that the correlation exclusively is associated to the 52 ppm resonance intensity. With the assumption that this resonance reflects methyl groups in choline/phosphatidyl choline, the intensity changes in the 52 ppm resonance imply alterations in membrane properties. Accordingly, the data indicate a relationship between membrane properties and the amount of water being expelled from muscle cells postmortem, which supports the hypothesis that disruption of membranes is implicated in the postmortem mobilization of muscle water.     

Natural variability in bovine milk oligosaccharides from Danish Jersey and Holstein-Friesian breeds

Free oligosaccharides are key components of human milk and play multiple roles in the health of the neonate, by stimulating growth of selected beneficial bacteria in the gut, participating in development of the brain, and exerting antipathogenic activity. However, the concentration of oligosaccharides is low in mature bovine milk, normally used for infant formula, compared with both human colostrum and mature human milk. Characterization of bovine milk oligosaccharides in different breeds is crucial for the identification of viable sources for oligosaccharide purification. An improved source of oligosaccharides can lead to infant formula with improved oligosaccharide functionality. In the present study we have analyzed milk oligosaccharides by high-performance liquid chromatography chip quadrupole time-of-flight mass spectrometry and performed a detailed data analysis using both univariate and multivariate methods. Both statistical tools revealed several differences in oligosaccharide profiles between milk samples from the two Danish breeds, Jersey and Holstein-Friesians. Jersey milk contained higher relative amounts of both sialylated and the more complex neutral fucosylated oligosaccharides, while the Holstein-Friesian milk had higher abundance of smaller and simpler neutral oligosaccharides. The statistical analyses revealed that Jersey milk contains levels of fucosylated oligosaccharides significantly higher than that of Holstein-Friesian milk. Jersey milk also possesses oligosaccharides with a higher degree of complexity and functional residues (fucose and sialic acid), suggesting it may therefore offer advantages in term of a wider array of bioactivities.     

Relationship between the metabolite profile and technological properties of bovine milk from two dairy breeds elucidated by NMR-based metabolomics

The aim of the present study was to investigate the relationship between the metabolite profile of milk and important technological properties by using nuclear magnetic resonance (NMR)-based metabolomics. The metabolomics approach was introduced for the metabolic profiling of a set of milk samples from two dairy breeds representing a wide span in coagulation properties. The milk metabolite profiles obtained by proton and carbon NMR spectroscopy could be correlated to breed and, more interestingly, also with the coagulation profile, as established by traditional methods by using principal component analysis (PCA). The metabolites responsible for the separation into breed could mainly be ascribed to carnitine and lactose, whereas the metabolites varying in the samples with respect to coagulation properties included citrate, choline, carnitine, and lactose. The results found in the present study demonstrated a promising potential of NMR-based metabolomics for a rapid analysis and classification of milk samples, both of which are useful for the dairy industry.     

The involvement of ethylene and calcium in gray mold of pelargonium,ruscus, and rose plants

Leaves ofPelargonium domesticum andRuscus hypoglossum infected byBotrytis cinerea Pers., produced 12 and 1.5 nl ethylene/h/g, respectively, 34 days after inoculation; wounded or healthy leaves and phyloclades of them produced much lower amounts. When the fungus was grown on dead leaves it produced negligible amounts of the gas even when supplemented with methionine. Exogenous ethylene enhanced gray mold development in both hosts. Silver thiosulfate, aminooxyacetic acid (AOA) and aminoethoxyvinylglycine significantly inhibited disease development in pelargonium, and the latter two compounds inhibited ethylene production. AOA inhibited disease development and ethylene production in cut rose flowers; calcium ions inhibited disease development whereas the chelator EGTA [ethylene glycol bis-(β-aminoethyl ether)N,N,N′,N′-tetraacetic acid] enhanced it. Disease suppression by an excess of Ca²⁺ was correlated with repression of ethylene production by the flowers, whereas deficiency in Ca²⁺ increased disease severity. Publication of the Agricultural Research Organization. No. 2413-E, 1988 series.     

Dynamic high-resolution 1H and 31P NMR spectroscopy and 1H T2 measurements in postmortem rabbit muscles using slow magic angle spinning

Postmortem changes in rabbit muscle tissue with different glycogen status (normal vs low) were followed continuously from 13 min postmortem until 8 h postmortem and again 20 h postmortem using simultaneous magic angle spinning (1)H and (31)P NMR spectroscopy together with measurement of the transverse relaxation time, T(2), of the muscle water. The (1)H metabolite spectra were measured using the phase-altered spinning sidebands (PASS) technique at a spinning rate of 40 Hz. pH values calculated from the (31)P NMR spectra using the chemical shifts of the C-6 line of histidine in the (1)H spectra and the chemical shifts of inorganic phosphate in the (31)P spectra confirmed the different muscle glycogen status in the tissues. High-resolution (1)H spectra obtained from the PASS technique revealed the presence of a new resonance line at approximately 6.8 ppm during the postmortem period, which were absent in muscles with low muscle glycogen content. This new resonance line may originate from the aminoprotons in creatine, and its appearance may be a result of a pH effect on the exchange rate between the amino and the water protons and thereby the NMR visibility. Alternatively, the new resonance line may originate from the aromatic protons in tyrosine, and its appearance may be a result of a pH-induced protein unfolding exposing hydrophobic amino acid residues to the aqueous environment. Further studies are needed to evaluate these hypotheses. Finally, distributed analysis of the water T(2) relaxation data revealed three relaxation populations and an increase in the population believed to reflect extramyofibrillar water through the postmortem period. This increase was significantly reduced (p < 0.0001) in samples from animals with low muscle glycogen content, indicating that the pH is controlling the extent of postmortem expulsion of water from myofibrillar structures. The significance of the postmortem increase in the amount extramyofibrillar water on the water-holding capacity was verified by centrifugation, which showed a reduced centrifugation loss in muscles with low preslaughter glycogen status (0.9 vs 1.9%, p = 0.07).