Assessment of qualitative and quantitative parameters of healthy thyroid gland in adult horses by magnetic resonance imaging

In humans and pets, magnetic resonance imaging (MRI) provides important adjunctive anatomic information about the thyroid and its adjacent structures. To the best of our knowledge, however, no studies have been reported regarding the use of MRI for thyroid evaluation in horses. The aim of this study was to obtain qualitative and quantitative parameters of healthy thyroid gland in adult horses by MRI. Eleven horses were submitted to thyroid MRI evaluation. The healthy thyroid of these animals was ensured by normal examinations performed before (free thyroxine and ultrasound) and after (cytology) to MRI. It was concluded that the dorsal spin-echo T1-weighted, fast spin-echo T2-weighted, sagittal STIR and a three-dimensional T1-weighted gradient echo were the sequences that best characterized the thyroid lobes. Qualitatively, there was a significant variation in the elliptical format of some lobes considering the sagittal plane, which oscillated between rounded and flattened conformation. Excellent reproducibility of the quantitative thyroid parameters obtained was verified. The thyroid gland total volume mean of observers was 14.31 cm³ (±0.69). In relation with comparisons between quantitative parameters of left and right lobes, only differences regarding their respective lengths were observed. Thus, we can infer that the difference in some thyroid lobes format can be explained by the significant variation identified between left and right lobes lengths.     

Induction of variability in chimeric Aster cordifolius `White Elegans' through somaclonal variation

A protocol was developed for plant regeneration from leaf explants (laminaand petiole) of Aster cordifolius cultivar `White Elegans', which ischimeric for chromosome number and the ligulate flower colour. Explantswere cultured on MS or Gamborg's B5 medium in presence of IAA, 2,4-Dand BAP, alone or in combination. The highest frequency of shootregeneration was obtained on B5 medium supplemented with 2,4-D(0.09 mg l^-1) and BAP (1.8 mg l^-1). After the invitro rooting stage and acclimatization, 90% of plantlets survived in thegreenhouse. The regeneration process occurred by indirect organogenesisas shown by cyto-histology. The regenerants maintained the originalchimerism for chromosome number. Pink flowers instead of theexpected white ones were obtained in all regenerants. Ahypothesis for the uniform colour change is discussed. Variations weredetected in the regenerant generation (R₀) for quantitative characterssuch as capitulum and disc diameters, and number of ligulate flowers. SinceAster can be vegetatively propagated, selection of variants in R₀can provide interesting material for breeding purposes.     

Increasing resistance to Fusarium crown and root rot in asparagus by gametophyte selection

In garden asparagus, Fusarium crown and root rot is the main cause of crop decline. Since chemical treatments are inefficient, efforts should focus on the development of resistant cultivars to control the disease. Toxic culture filtrate (TCF) of F. oxysporum has affected asparagus pollen germination and tube growth. Consequently, gametophyte selection was evaluated to ascertain if the application of selective agents at this level could increase selection efficiency. Two susceptible pistillate plants and one tolerant and one susceptible staminate plants were used in controlled crosses. Before pollination, a drop of a germination vehicle with TCF or without it was applied to the stigmas. Some pollinated pistils were fixed and analyzed by fluorescence microscopy; the rest were left on the plant for seed production. Fifty to 200 seeds were obtained per treatment combination (staminate plant x pistillate plant x pollination vehicle). The derived plantlets were inoculated in vitroand evaluated for disease symptoms. The application of TCF to stigmas reduced pollen germination and tube growth compared with untreated controls,regardless of the genotypic combination. Pollen germination and tube growth was poorer for the tolerant staminate genotype than for the susceptible one. When the TCF was applied, the number of seeds per pollination in comparison with the controls diminished only when the susceptible genotype was the pollinator. The percentage of affected root area of the progenies obtained after applying the TCF was lower than in the controls only when the tolerant genotype was the pollinator. Increasing Fusarium resistance in asparagus by means of gametophyte selection seems feasible. This revised version was published online in August 2006 with corrections to the Cover Date.     

Ovum pick-up and in vitro maturation in spotted paca (Cuniculus paca—Linnaeus, 1766)

We tested FSHp, eCG and FSHp + eCG to establish ovum pick-up (OPU) and in vitro maturation method in spotted paca. Eight healthy adult females were subjected to each of four treatments to stimulate ovarian follicular growth. All females were subjected to a hormonal protocol using a single dose of 45 mg of injectable progesterone and single intramuscular injection of 0.075 mg d-cloprostenol on day 6. Ovarian stimulation was carried out as follows: in Group TFE (FSHp and eCG), animals were treated with a single dose of 80 mg of FSHp and 200 IU of eCG intramuscularly on day 6 after the application of progesterone; in Group TF (FSHp), they were treated with a single dose of 80 mg of FSHp intramuscularly on day 6 after application of progesterone; in Group treatment eCG, they were treated with 200 IU of eCG intramuscularly on day 6 after application of progesterone; and in Group TC (saline solution), 1 ml of saline solution was administered to control does. The OPU was performed between 22 and 26 hr after gonadotropin treatments. All recovered oocytes were placed into maturation media and incubated for 24 hr. There were no differences among the mean number of observed follicles, aspirated follicles and oocytes recovered per treatment. Oocyte maturation rates did not differ among groups, except, TF and treatment eCG oocytes had greater maturation rates than TC oocytes. In this study, gonadotropin administration failed to superovulate treated does and increase oocyte retrieval efficiency. Despite the feasibility of the procedure, further studies are needed to develop and refine hormonal protocols for oocyte recovery and in vitro maturation in this species.     

Novel approach to study oxidative stability of extra virgin olive oils: importance of alpha-tocopherol concentration

The stability of extra virgin olive oils is mainly due to their relatively low fatty acids unsaturation and to the antioxidant activity of some of the unsaponifiable components. We studied the activity of alpha-tocopherol in extra virgin olive oil in its natural state, using new and simple oxidizing conditions in "thin layer" and "bulk phase". Oxidation time course was monitored at 37 degrees C or 75 degrees C. A storage test was also performed. Two parameters were evaluated: depletion of alpha-tocopherol and formation of PUFA hydroperoxides, measured as conjugated dienes (CD) and peroxide value. The value of complex polyphenols was also measured. alpha-Tocopherol concentration decreased in function of time and temperature and showed a strong inverse correlation with the increase of CD. When alpha-tocopherol reached a "threshold value" of 60-70 mg/kg, a significant increase of CD formation was observed, together with a good correlation between CD and peroxide value. The initial amount of alpha-tocopherol is one of the components that influences oil oxidative susceptibility.     

Differential immunohistochemical expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in cow uteri with adenomyosis during follicular phase

The present investigation was intended to show a different immunohistochemical profile of matrix metalloproteinase-2 and Tissue inhibitor metalloproteinase-2 in bovine uteri with adenomyosis during follicular phase. Uterine samples of 32 cows in reproductive age were taken from the medial third of one of the uterine horns and grouped according to the adenomyosis degree (superficial and deep). Tissue sections (4 μm) were incubated overnight at 4°C with monoclonal antibody for matrix metalloproteinase-2 and Tissue inhibitor metalloproteinase-2. Staining intensities were evaluated in the luminal epithelium, ectopic and dystopic endometrial tissue (stroma, capillaries and glands), endometrial-myometrial border, myometrium, myometrial vessels (middle tunic and endothelium). The matrix metalloproteinase-2 expression was higher for deep adenomyosis samples, showing a differential mean reactivity in superficial endometrium, myometrial vessels, myometrium adjacent to adenomyotic focus and endometrial-myometrial border (P < 0.05). Moreover, matrix metalloproteinase-2 expression was higher in deep adenomyosis samples than that of Tissue inhibitor metalloproteinase-2 in almost all uterine structures analyzed (except for the endometrial and myometrial vessels and endometrial-myometrial border). The opposite was observed in the follicular phase, for both normal specimens and with superficial adenomyosis, where Tissue inhibitor metalloproteinase-2 expression was higher than that of matrix metalloproteinase-2. In conclusion, a differential pattern of matrix metalloproteinase-2 and Tissue inhibitor metalloproteinase-2 was observed in cow uteri with adenomyosis.