Effect of oxytocin on expression of cytosolic phospholipase A2 mRNA and protein in ovine endometrial tissue in vivo

The induction of endometrial prostaglandin (PG) F₂ synthesis by oxytocin is dependent upon activation of phospholipase (PL) A₂ and mobilization of arachidonic acid. The objective of this study was to determine if oxytocin stimulates PGF₂ synthesis by inducing synthesis of cytosolic PLA₂ (cPLA₂). In Experiment 1, 15 ovariectomized ewes were given progesterone and estradiol to simulate an estrous cycle. Ewes were then given an injection of oxytocin on Day 14 of the simulated estrous cycle. Jugular blood samples were collected and assayed for 13,14-dihydro-15-keto-prostaglandin F₂ (PGFM). Uteri were collected at 0, 7.5, 25, 90, or 240 min postinjection (n = 3 ewes/time point). Total RNA was isolated from caruncular endometrium and subjected to dot-blot analysis. Oxytocin induced a rapid and transient increase in serum PGFM (P < 0.01). However, endometrial concentrations of cPLA2 mRNA did not change following oxytocin administration (P > 0.10). In Experiment 2, 11 ovary-intact ewes were given oxytocin (n = 5) or saline (n = 6) on Day 15 after estrus. Jugular blood samples were collected and assayed for serum concentrations of PGFM. Uteri were collected at 15 min postinjection. Homogenates were prepared from caruncular endometrium and subjected to Western blot analysis. Concentrations of PGFM were higher in oxytocin treated ewes compared to saline treated ewes at 15 min postinjection (P < 0.01). Endometrial concentrations of cPLA₂ protein were greater in the cytosolic than in the microsomal fraction (P < 0.01). Oxytocin did not affect the amount of cPLA₂ protein in either fraction (P > 0.10). In conclusion, oxytocin did not effect expression of either cPLA₂ mRNA or protein in ovine endometrium. Oxytocin may stimulate PGF₂ synthesis by activating cPLA₂ protein that is already present in an inactive form.     

FEASIBILITY FOR USING HYPOFRACTIONATED STEREOTACTIC VOLUMETRIC MODULATED ARC RADIOTHERAPY (VMAT) WITH ADAPTIVE PLANNING FOR TREATMENT OF THYMOMA IN RABBITS: 15 CASES

Thymoma is a relatively common tumor in rabbits. Treatment with surgery, radiation therapy, and chemotherapy alone or in combination has been reported with varying outcomes. Stereotactic volumetric modulated arc radiotherapy delivered in a hypofractionated manner allows high doses of radiation to be delivered to the target volume while allowing sparing of adjacent critical structures. This therapy is ideally suited for thymomas in rabbits given their size, the difficulty of multiple anesthesia episodes and the complexity of the radiotherapy plans required due to the tumor's proximity to the heart, lungs, and mediastinal structures. Fifteen rabbits with thymoma were prospectively recruited for this observational, single institution, single arm clinical study. All rabbits were imaged with both computed tomography (CT) and magnetic resonance imaging (MRI). A total dose of 40 Gy in six fractions was delivered using a single arc over an 11‐day period with repeat CT simulation done every other fraction for adaptive planning. Follow‐up evaluation was done through repeat CT and MRI imaging and revealed complete responses using the Response Evaluation Criteria in Solid Tumors (RECIST) criteria. Two rabbits had died at 618 and 718 days, 10 were alive and three were lost to follow‐up. Observed acute and late effects were graded according to the Veterinary Radiation Therapy Oncology Group (VRTOG) criteria and were found to be minimal.     

Response of gladiolus (Gladiolus spp) plants after exposure corms to chitosan and hot water treatments

The state of Morelos, Mexico has gradually become an important producer of gladiolus. Some preconditioning treatments of corms are empirically done causing uneven emergence and low quality of flowers. In this investigation, before planting, gladiolus corms var. ‘Blanca Borrego’ were dipped in chitosan (chitosan reagent and commercial chitosan Biorend^®), in hot water at various temperatures and in treatments combined with Biorend^® at 1.5% and hot water. Results indicated that the most influenced variables were corm germination, number of flowers per spike, number of cormlets and vase life. Overall, the commercial product Biorend^® at 1.5% accelerated corm emergence in approximately 4 days, the number of flowers increased by 2–7 and the vase life extended for 3 days. The number of cormlets was also duplicated. Corms dipped in the commercial chitosan Biorend^® at 1.5% at different intervals of time were not greatly affected except for the emergence and number of cormlets. However, for this experiment there were significant effects on the number of leaves and flowers because of the interactions between chitosan and the immersion time. The temperature of 55 °C affected plant development because emergence was delayed by 6 days; and there were less number of leaves, flowers and cormlets. On the other hand, the incidence of Fusarium oxysporum in naturally infected corms was 0% at temperatures of 55 °C and 50 °C. Immersion times (0, 10, 15 and 20 min) in hot water at 50 °C did not show significant effects on plant development and vase life. Corms dipped in Biorend^® at 1.5% and hot water at 50 °C accelerated their emergence for about 1–7 days, the number of flowers increased by two, extended the storage life for 1–3 days and increased the number of cormlets. The integration of these two treatments -Biorend^® and hot water- might be a good option for increasing the gladiolus plant quality and vase life.     

Cover Image,Volume 14, Issue 3

The cover image, by Giuliano Bettini et al., is based on the Original Article Cytological grading of canine cutaneous mast cell tumours, DOI: 10.1111/vco.12090 .

     

A rare Cryptosporidium parvum genotype associated with infection of lambs and zoonotic transmission in Italy

An outbreak of cryptosporidiosis occurred in a mixed sheep/cattle farm of Central Italy in October 2011. A total of 450 ovines (250 sheep and 200 lambs) and 140 bovines (130 cows and 10 calves) were housed in two separated units, at the time of the outbreak. About half of the lambs had diarrhea due to Cryptosporidium sp. with a mortality rate of 80%; calves were not infected. Genomic DNA was extracted from an archived slide and from fecal specimens, and the parasite was identified as Cryptosporidium parvum by PCR and sequence analysis at the CpA135 gene. Genotyping at the GP60 gene showed the presence of a very rare genotype, IIaA20G2R1. Shortly after the outbreak was identified, the son of the farm's owner, aged 18 months, experienced an acute gastroenteritis and was hospitalized due to recurrent episodes of diarrhea, fever, vomiting and lack of appetite. The feces tested negative for bacteria and viruses, whereas cryptosporidiosis was diagnosed by microscopy and an immunochromatographic test. Molecular typing identified the C. parvum genotype IIaA20G2R1 in the feces of the child. This is the first case of transmission of cryptosporidiosis in Italy involving lambs as source of oocysts infectious to humans.     

COMPARISONS AMONG COMPUTED TOMOGRAPHIC FEATURES OF ADIPOSE MASSES IN DOGS AND CATS

A better understanding of the CT features of different forms of canine and feline adipose tumors would be valuable for improving patient management and treatment. The purpose of this retrospective, cross‐sectional study was to describe and compare the CT features of pathologically confirmed lipomas, infiltrative lipomas, and liposarcomas in a sample of canine and feline patients. A total of 50 animals (46 dogs, four cats) and a total of 60 lesions (23 lipomas, 20 infiltrative lipomas, and 17 liposarcomas) were included in the study. Lipomas appeared as round to oval‐shaped (n = 21), well‐marginated (n = 20) fat‐attenuating lesions. Infiltrative lipomas appeared as homogeneous, fat‐attenuating masses but, unlike lipomas, they were most commonly characterized by an irregular shape (75%; P < 0.001), and linear components, hyperattenuating relative to the surrounding fat (100%; P < 0.05). Liposarcomas were represented exclusively by heterogeneous lesions with soft tissue attenuating components with a multinodular appearance (76.5%; P < 0.05). Regional lymphadenopathy (n = 10) and amorphous mineralization (n = 4) were also observed in association with liposarcomas. Computed tomography can provide useful information regarding disease location, extent, and involvement of the adjacent structures. Tumor definition and shape were the most useful parameters to differentiate between lipomas and infiltrative lipomas. The presence of a heterogeneous mass, with a multinodular soft tissue component and associated regional lymphadenopathy and mineralization, were features favoring a diagnosis of liposarcoma.     

Dual-label time-resolved fluoroimmunoassay for simultaneous quantification of haptoglobin and C-reactive protein in meat juice from pigs

A new method was developed to simultaneously measure 2 acute-phase proteins (APPs) by time-resolved immunofluorometry. The assay, based on double-label quantification of haptoglobin (Hp) and C-reactive protein (CRP) in meat juice samples from pigs, was constructed by use of a combination of europium and samarium chelate lanthanides as labels. Meat juice samples from 154 pigs were used for analytic and clinical validation of the assay through determination of precision, accuracy, limit of detection, and quantification. The analytic performance of the assay was satisfactory, with good intra-assay and interassay precision and accuracy. The levels of Hp and CRP were increased in the meat juice samples of diseased animals compared with healthy ones. According to the results, higher sensitivity could be achieved if the cut-off values of both proteins were taken into account for clinical relevance rather than used individually. Since the dual assay saved both time and sample, it could be used as a rapid and sensitive screening test in porcine production.     

Synthesis and Antifouling Activity Evaluation of Analogs of Bromosphaerol,a Brominated Diterpene Isolated from the Red Alga Sphaerococcus coronopifolius

Marine biofouling is an epibiotic biological process that affects almost any kind of submerged surface, causing globally significant economic problems mainly for the shipping industry and aquaculture companies, and its prevention so far has been associated with adverse environmental effects for non-target organisms. Previously, we have identified bromosphaerol (1), a brominated diterpene isolated from the red alga Sphaerococcus coronopifolius, as a promising agent with significant antifouling activity, exerting strong anti-settlement activity against larvae of Amphibalanus (Balanus) amphitrite and very low toxicity. The significant antifouling activity and low toxicity of bromosphaerol (1) motivated us to explore its chemistry, aiming to optimize its antifouling potential through the preparation of a number of analogs. Following different synthetic routes, we successfully synthesized 15 structural analogs (2–16) of bromosphaerol (1), decorated with different functional groups. The anti-settlement activity (EC₅₀) and the degree of toxicity (LC₅₀) of the bromosphaerol derivatives were evaluated using cyprids and nauplii of the cirriped crustacean A. amphitrite as a model organism. Derivatives 2, 4, and 6–16 showed diverse levels of antifouling activity. Among them, compounds 9 and 13 can be considered as well-performing antifoulants, exerting their activity through a non-toxic mechanism.     

Effect of food on the pharmacokinetics of oral cefuroxime axetil in dogs

Cefuroxime axetil pharmacokinetic profile was investigated in 12 Beagle dogs after single intravenous and oral administration of tablets or suspension at a dose of 20 mg/kg, under both fasting and fed conditions. A three-period, three-treatment crossover study (IV, PO under fasting and fed condition) was applied. Blood samples were withdrawn at predetermined times over a 12-hr period. Cefuroxime plasma concentrations were determined by HPLC. Data were analyzed by compartmental analysis. No statistically significant differences were observed between formulations and feeding conditions on PK parameters. Independently of the feeding condition, absorption of cefuroxime axetil after tablet administration was low and erratic. The drug has been quantified in plasma in 3 out of 6 and 5 out of 6 dogs in the fasted and fed groups. For this formulation, the bioavailability (F), peak plasma concentration (C_max), and area under the concentration–time curve (AUC) of cefuroxime axetil were significantly enhanced (p < .05) by the concomitant ingestion of food (32.97 ± 13.47–14.08 ± 7.79%, 6.30 ± 2.62–2.74 ± 0.66 µg/ml, and 15.75 ± 3.98–7.82 ± 2.76 µg.hr/ml for F, C_max, and AUC in fed and fasted dogs, respectively), while for cefuroxime axetil suspension, feeding conditions affected only the rate of absorption, as reflected by the significantly shorter absorption half-life (T_½(a)) and time to peak concentration (T_max) (0.55 ± 0.27–1.15 ± 0.19 hr and 1.21 ± 0.22–1.70 ± 0.30 for T_½(a) and T_max in fed and fasted dogs, respectively). For cefuroxime axetil tablets, T > MIC (≤1 µg/ml) was <2 hr in fasted and ≈4 hr in fed animals, and for cefuroxime axetil suspension, T > MIC (≤1 µg/ml) was ≈5 hr and for T >MIC (≤4 µg/ml) was ≈2.5 hr for fasted and fed dogs, respectively. Cefuroxime axetil as a suspension formulation seems to be a better option than tablets. However, its short permanence in plasma could reduce its clinical usefulness in dogs.