Development and Validation of a UPLC-MS/MS Method to Monitor Cephapirin Excretion in Dairy Cows following Intramammary Infusion

Cephapirin, a cephalosporin antibiotic, is used by the majority of dairy farms in the US. Fecal and urinary excretion of cephapirin could introduce this compound into the environment when manure is land applied as fertilizer, and may cause development of bacterial resistance to antibiotics critical for human health. The environmental loading of cephapirin by the livestock industry remains un-assessed, largely due to a lack of appropriate analytical methods. Therefore, this study aimed to develop and validate a cephapirin quantification method to capture the temporal pattern of cephapirin excretion in dairy cows following intramammary infusion. The method includes an extraction with phosphate buffer and methanol, solid-phase extraction (SPE) clean-up, and quantification using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The LOQ values of the developed method were 4.02 µg kg⁻¹ and 0.96 µg L⁻¹ for feces and urine, respectively. This robust method recovered >60% and >80% cephapirin from spiked blank fecal and urine samples, respectively, with acceptable intra- and inter-day variation (<10%). Using this method, we detected trace amounts (µg kg⁻¹) of cephapirin in dairy cow feces, and cephapirin in urine was detected at very high concentrations (133 to 480 µg L⁻¹). Cephapirin was primarily excreted via urine and its urinary excretion was influenced by day (P = 0.03). Peak excretion (2.69 mg) was on day 1 following intramammary infusion and decreased sharply thereafter (0.19, 0.19, 0.08, and 0.17 mg on day 2, 3, 4, and 5, respectively) reflecting a quadratic pattern of excretion (Quadratic: P = 0.03). The described method for quantification of cephapirin in bovine feces and urine is sensitive, accurate, and robust and allowed to monitor the pattern of cephapirin excretion in dairy cows. This data will help develop manure segregation and treatment methods to minimize the risk of antibiotic loading to the environment from dairy farms.     

Detection of a calmodulin-sensitive cyclic nucleotide phosphodiesterase in rat parotid gland

Calmodulin coupled to Sepharose has provided a rapid and sensitive means of isolating a cyclic nucleotide phosphodiesterase activity which is stimulated by the calmodulin-Ca²⁺ complex, from rat parotid gland. Initial experiments established that phosphodiesterase activity sensitive to calmodulin and Ca²⁺ could not be demonstrated in crude extracts of rat parotid gland or after partial purification of rat parotid phosphodiesterase over DEAE-cellulose. However, it was possible to readily demonstrate the presence of a cyclic nucleotide phosphodiesterase activity regulated by calmodulin if the extracts were first purified by batch ion-exchange chromatography over DEAE-cellulose followed by affinity chromatography with calmodulin coupled to Sepharose. The batch ion-exchange chromatography step removed the major portion of free parotid calmodulin which could compete with calmodulin-coupled Sepharose for the proteins regulated by calmodulin. Thus, by employing an initial chromatography step over DEAE-cellulose to separate phosphodiesterase activity from calmodulin, it was possible to increase the recovery of calmodulin-sensitive phosphodiesterase after affinity chromatography with calmodulin coupled to Sepharose. This approach should be useful for demonstrating the presence of and for purifying other parotid proteins regulated by calmodulin.     

Power frequency magnetic field exposures among nurses in a neonatal intensive care unit and a normal newborn nursery

Given the current interest in potential carcinogenic and developmental effects of exposure to extremely-low-frequency electromagnetic fields, there is a need to identify cohorts of exposed female workers for future epidemiologic investigations. This study was designed to test the hypothesis that nurses working in neonatal intensive care units (NICU) may be significantly exposed to power-frequency magnetic fields. An electromagnetic field monitor was used to measure magnetic fields at distances of 5, 15, 30, and 60 cm from the surfaces of each device used in the NICU. Six female nurses assigned to the NICU (the “exposed” group) and six female nurses working in the normal newborn nursery (the “referent” group) wore EMDEX dosimeters for the entire duration of their 12 h shifts. An investigator kept a detailed log of each NICU subject's whereabouts for the first one-third of her shift. Magnetic fields at 5 cm from the front (defined by the nurses' usual work area) of the NICU devices ranged from less than 0.1 to 114 μT and in all cases decreased considerably with increasing distance. The geometric mean of the shift-time-weighted average exposure of the NICU nurses was 0.17 μT compared with 0.11 μT for the normal newborn nurses. The percentage of time when subjects were exposed to magnetic fields of 0.4 μT or greater ranged from 5.8% to 15.6% for the NICU nurses, 0.4% to 2.9% for five of the comparison group nurses, and was 9.4% for one of the normal newborn nurses with unidentified aberrantly high exposures. Log data revealed that the vast majority of observed peaks among NICU nurses occurred while subjects were in close proximity to infant bed units. We conclude that NICU nurses represent one female-intensive job sector with intermittent high exposures to ELF magnetic fields and encourage larger exposure studies of nurses in a variety of medical settings. © 1994 Wiley-Liss, Inc.     

BIOCHEMICAL, HEMATOLOGIC AND COAGULATION VALUES IN THE HEREFORD CALF

Biochemical, hematologic and coagulation values have been determined in normal Hereford calves of both sexes, two to six months of age. These data generally agree with those of others and indicate a similarity in calves irrespective of their age, sex or breed. Prothrombin times are markedly longer in the calf than in man. This has resulted in alterations of anticoagulation regimens after implantation of mechanical circulatory support systems.     

The distance dependence prediction of the Janzen-Connell hypothesis: a meta-analysis

The Janzen-Connell hypothesis explains the maintenance of tropical diversity through the interacting effects of parent-centered dispersal patterns and distance- and density-dependent propagule survival. These effects were thought to support regular spacing of species within tropical forest, enhancing diversity. One of the predictions of the hypothesis is that seed and seedling survival should improve with increased parental distance. Although there are many independent tests of this hypothesis for individual species, there are few synthetic studies that have brought these data together to test its validity across species. This paper reports the results of a meta-analysis of the effect of distance on enhancing propagule survival, employing an odds-ratio effect size metric. We found no general support for the distance-dependent prediction of the hypothesis, and conclude that further testing to explore this hypothesis as a diversity-maintaining mechanism is unnecessary. However, we did find that distance from parent slightly reduces survivorship in the temperate zone, as contrasted with the tropics, and we saw stronger evidence in support of the hypothesis for seedlings than for seeds. The phenomenon of enhanced propagule survival with distance from the parent may be important for the population biology of particular species, but it is not a general phenomenon across communities, life history stages or life forms.     

Nucleic acids in relation to cell division in Lilium longiflorum

Methods are described for preparing cell suspensions of Lilium microsporocytes, microspores and pollen grains; for obtaining cell counts of these suspensions; and for their analysis for pentose nucleic acid (PNA) and desoxypentose nucleic acid (DNA).The results of these analyses have been calculated to nucleic acid content in μμg per microsporocyte, microspore or pollen grain, and the results related to logarithm of flower bud length, an index of the developmental status of the cells, and of their temporal relationship to meiosis, microspore mitosis and opening of the flower.DNA content per cell drops sharply at the end of meiosis, with the formation of four microspores from each microsporocyte. It then increases gradually during the microspore interphase between meiosis and the microspore mitosis. At microspore mitosis DNA content doubles rapidly. In the development of the resulting binucleate pollen grain, from microspore mitosis until the opening of the flower, there is a further gradual increase of DNA content. PNA content of these cells follows the same pattern up to microspore mitosis at a level about twice that of DNA, increases sharply at mitosis, and continues to increase rapidly at a rate nine times that for DNA in the maturing pollen grain.The absolute amounts of DNA and PNA are great. At the time of anthesis the two-celled pollen grain contains about 375 μμg of DNA and 1705 μμg of PNA.