A simple and sensitive liquid chromatography-mass spectrometry confirmatory method for analyzing sulfonamide antibacterials in milk and egg

A simple and specific method able to identify and quantify traces of 14 sulfonamide antibacterials (SAs) in milk and eggs is presented. This method uses a single solid-phase extraction (SPE) cartridge for simultaneous extraction and purification of SAs in the above matrices. Milk and egg samples are passed through a Carbograph 4 sorption cartridge. After analyte desorption, an aliquot of the final extract is injected into a liquid chromatography-mass spectrometry (LC-MS) instrument equipped with an electrospray ion source (ESI) and a single quadrupole. MS data acquisition is performed in the positive-ion mode and by a time-scheduled multiple-ion selected ion monitoring program. Compared to two published methods, the present protocol extracted larger amounts of SAs from both milk and egg and decreased the analysis time by a factor of 3 with milk samples and by a factor of 2 with egg samples. Recovery of SAs in milk at the 5 ppb level ranged between 76 and 112% with relative standard deviations (RSDs) of 相似文献   

Isotretinoin in the treatment of multiple benign pilomatrixomas in a mixed-breed dog

An 18-month-old cross-breed dog developed multiple skin nodules, which over 8 months had increased in size and number. Biopsy samples were submitted for histopathological examination and revealed multiple benign pilomatrixomas. The dog was treated with 1 mg kg(-1) isotretinoin daily, which led to a stabilization of the size and number of the lesions. Attempts to lower the dosage to an every other day regimen led to an increase in the nodules' size and number. Monthly complete blood count and chemistry tests and Schirmer tear tests were performed to monitor any adverse effects of the retinoid treatment. No adverse effects were noted. To the authors' knowledge, this is the first report of multiple pilomatrixomas arising in a young dog and successfully controlled with oral retinoids.     

Reconstruction of the lower eyelid by third eyelid lateral advancement and local transposition cutaneous flap after "en bloc" resection of squamous cell carcinoma in 5 cats

OBJECTIVE: To describe a novel technique for blepharoplasty to cover a tissue defect involving >/=50% of the lower eyelid. STUDY DESIGN: Prospective clinical study. ANIMALS: Five cats with lower eyelid squamous cell carcinoma (SCC). METHODS: En bloc resection of SCC by removing >/=50% of the lower lid with either the medial or lateral canthus was performed without other adjunctive treatment for SCC. The lid defect was reconstructed with a transposition skin flap derived from the frontal (medial defect) or temporal (lateral defect) region. The third eyelid was advanced laterally without dissection from its insertion; its outer conjunctival layer was removed, and the skin flap was sutured with single interrupted sutures dorsally over the nictitating membrane, ventrally to the cutaneous edge of the surgical wound and medially or laterally (depending on the canthus removed) to the skin of the remaining lower lid. RESULTS: Satisfactory cosmetic and functional results were achieved and the Schirmer tear tests were normal. In 2 cats, the skin flap needed monthly hair trimming to avoid corneal lesions. CONCLUSIONS: After en bloc resection of SCC involving >/=50% of the lower eyelid, reconstruction can be achieved by relocation of the third eyelid and use of a cutaneous transposition flap sutured to the scarified external surface of the third eyelid. Eyelid apposition and lacrimal function were preserved. CLINICAL RELEVANCE: Blepharoplasty using a cutaneous transposition flap sutured to the scarified surface of a relocated third eyelid should be considered for reconstruction of lower eyelid defects with >/=50% tissue loss of the lid margin.     

Primary hyperoxaluria (L-glyceric aciduria) in a cat

A 7-month-old, male European cat was examined because of weakness and inappetence. The cat was dehydrated, polypnoeic and severely weak. Severe, generalised muscle atrophy was present. Spinal reflexes were all decreased to absent. Blood analysis and urinalysis showed several abnormalities, including intermittent hyperoxaluria. The L-gliceric acid concentration was remarkably increased. Electrodiagnostic tests of the peripheral nervous system were abnormal. At necropsy, generalised muscle atrophy was observed. Microscopically, both kidneys showed intraluminal birefringent oxalate crystals. Motor neuron degeneration and accumulation of neurofilaments were observed in the axons of the spinal motor neurons.     

Procalcitonin gene expression after LPS stimulation in the porcine animal model

Procalcitonin (PCT), recognised as a marker of sepsis, was investigated in a porcine model of endotoxic shock. The results showed that continuous IV infusion (1-4 h) of LPS (40 μg/kg) in pigs was able to induce a generalised increase of PCT expression in lung, heart, kidney and liver. The increase in PCT was significant only in kidney and was accompanied by an increase in IL-6 gene expression. In vitro results demonstrated that peripheral blood mononuclear cells (PBMCs), as well as endothelial cells, were potentially capable of contributing to in vivo extrathyroidal PCT production. These findings support previous data from pigs concerning the occurrence of widespread activation of PCT extrathyroidal gene expression during endotoxic shock in pigs. Nevertheless, the levels of PCT detected were very low, suggesting the need for additional studies to validate the pig as a reliable animal model for investigating the role of PCT in sepsis.     

Genomic characterization of H1N2 swine influenza viruses in Italy

Three subtypes (H1N1, H1N2, and H3N2) are currently diffused worldwide in pigs. The H1N2 subtype was detected for the first time in Italian pigs in 1998. To investigate the genetic characteristics and the molecular evolution of this subtype in Italy, we conducted a phylogenetic analysis of whole genome sequences of 26 strains isolated from 1998 to 2010. Phylogenetic analysis of HA and NA genes showed differences between the older (1998-2003) and the more recent strains (2003-2010). The older isolates were closely related to the established European H1N2 lineage, whereas the more recent isolates possessed a different NA deriving from recent human H3N2 viruses. Two other reassortant H1N2 strains have been detected: A/sw/It/22530/02 has the HA gene that is closely related to H1N1 viruses; A/sw/It/58769/10 is an uncommon strain with an HA that is closely related to H1N1 and an NA similar to H3N2 SIVs. Amino acid analysis revealed interesting features: a deletion of two amino acids (146-147) in the HA gene of the recent isolates and two strains isolated in 1998; the presence of the uncommon aa change (N66S), in the PB1-F2 protein in strains isolated from 2009 to 2010, which is said to have contributed to the increased virulence. These results demonstrate the importance of pigs as mixing vessels for animal and human influenza and show the presence and establishment of reassortant strains involving human viruses in pigs in Italy. These findings also highlighted different genomic characteristics of the NA gene the recent Italian strains compared to circulating European viruses.     

Novel Prunus rootstock somaclonal variants with divergent ability to tolerate waterlogging

Plants require access to free water for nutrient uptake, but excess water surrounding the roots can be injurious or even lethal because it blocks the transfer of free oxygen between the soil and the atmosphere. Genetic improvement efforts in this study were focused on the increased tolerance in roots to waterlogging. Among a pool of clones generated in vitro from leaf explants of rootstock Mr.S.2/5 of Prunus cerasifera L., the S.4 clone was flood tolerant whereas the S.1 clone was sensitive. The S.4 clone formed adventitious roots on exposure to flooding. Moreover, the chlorophyll content and mitochondrial activity in the leaf and root, soluble sugar content, alcohol dehydrogenase activity and ethylene content were different between the clones. The sorbitol transporter gene (SOT1) was up-regulated during hypoxia, the alcohol dehydrogenase genes (ADH1 and ADH3) were up-regulated in the leaves and down-regulated in the roots of the S.4 clone during hypoxia, and the 1-aminocyclopropane-1-oxidase gene (ACO1) was up-regulated in the leaves and roots of the S.4 clone during hypoxia and down-regulated in the wild-type roots. In addition, in the S.4 root, hypoxia induced significant down-regulation of a glycosyltransferase-like gene (GTL), which has a yet-undefined role. Although the relevant variation in the S.4 genome has yet to be determined, genetic alteration clearly conferred a flooding-tolerant phenotype. The isolation of novel somaclonals with the same genomic background but with divergent tolerance to flooding may offer new insights in the elucidation of the genetic machinery of resistance to flooding and aid in the selection of new Prunus rootstocks to be used in various adverse environments.     

Seasonal variation in nitrogen isotopic composition of bog plant litter during 3 years of field decomposition

In this study, we describe the seasonal variation in ¹⁵N abundance in the litter of two Sphagnum species and four vascular plant species during 3 years of field decomposition in an Italian Alpine bog. Litter bags were periodically retrieved at the end of summer and winter periods, and the δ¹⁵N in residual litter was related to mass loss, litter chemistry, and climatic conditions. In Sphagnum litter, higher rates of decomposition during summer months were associated with an increase of δ¹⁵N probably due to the incorporation of microbial organic compounds rich in ¹⁵N. The litter of Eriophorum vaginatum and Carex rostrata was characterized by a decrease of δ¹⁵N, so that the final signature was significantly lower than in initial litter. On the other hand, the residual litter of Potentilla erecta and Calluna vulgaris was characterized by a final δ¹⁵N higher than in initial litter. Our data reported a seasonality of ¹⁵N abundance in the residual litter of Sphagnum species, but not in that of vascular plant species, thus highlighting the role of differences in litter chemistry.