Immunohistochemical Expression of Cyclooxygenase‐2 (COX‐2) in Feline Endometrial Adenocarcinoma and in Normal and Hyperplastic Endometria

Cyclooxygenase‐2 (COX‐2) is overexpressed in several human and animal neoplasms, including the human endometrial carcinoma. It has been suggested as a prognostic marker and a potential therapeutic target. This study aimed to (i) clarify histological aspects of feline endometrial adenocarcinomas (FEA) of the papillary serous type and (ii) characterize COX‐2 immunohistochemical expression in normal, hyperplastic and neoplastic endometrium in this species. Archived paraffin‐embedded tissue samples of 33 FEA, eight cystic endometrial hyperplasias (CEHs) and 21 samples of normal, healthy endometrium in the follicular (FS; n = 10) and luteal (LS; n = 11) stages were evaluated. Histological evaluation of haematoxylin and eosin‐stained sections of the FEA revealed a papillary proliferation of neoplastic cells of serous type, accompanied by clear and multinucleated cells. Other architectural arrangements mainly included solid and tubular growth. Randomly distributed areas of necrosis within the tumours were commonly observed. Invasion of the myometrium, of the serosa and of the vascular and/or lymphatic vessels was not constant features. The mean number of mitoses was higher in FEA compared to non‐neoplastic endometrium. COX‐2 scores were lower in FEA (p = 0.003) and CEH (p = 0.05) when compared to normal epithelium (NE). The loss of the membrane apical reinforcement in epithelial cells was observed in FEA samples, which was accompanied by the dislocation of COX‐2 labelling into the cytoplasm and the perinuclear area; in contrast, in epithelial cells in the healthy and hyperplastic endometria, the immunoreaction showed the characteristic pattern of apical membrane reinforcement, suggestive of the membrane polarization. COX‐2 epithelial scores were higher in the FS than in the LS. No differences were found in stromal COX‐2 expression between normal, CEH and FEA groups, but it was higher in the LS than in the FS. In summary, loss of COX‐2 compartmentalization in neoplastic epithelial cells might be one of the molecular events underlying endometrial carcinogenesis.     

Venous hepatic segmentation in dogs (Canis lupus familiaris—L. 1758)

The external shape of the liver is varied and determines specific vascular arrangements. This morphological relationship is important to establish hepatic segmentation in different species submitted to surgeries that aim to preserve a larger area of liver parenchyma. After observing 60 livers injected with Neoprene Latex and three plastic moulds obtained by corrosion, eight hepatic venous segments were identified, drained by six hepatic veins agrouped into segmental veins, which drained one sector (segments I, VI, VII and VIII) and intersegmental veins, which drained more than one sector (segments II/III and IV/V). They were described as follows: left intersegmental vein, formed by a segmental vein from the papillary process (segment I), two to three lateral left segmental veins that drained the segment II, and one to five left paramedian segmental veins that drained the segment III; sagittal intersegmental vein, formed by the confluence between segmental vein of the quadrate lobe (segment IV) and the medial right paramedian segmental vein, which derived from the segment V; lateral right paramedian vein drained the dorsocranial sector of the segment VI; the lateral right segmental vein, formed by one to four vessels that drained segment VII, and the segmental vein of the caudate process, which drained the segment VIII. Understanding the number and disposition of the hepatic veins in lobate livers is essential to reduce bleeding risks in surgeries. The nomenclature based on segmentation analogy of non-lobate liver could be less confusing and, therefore, be more useful in the surgical approaches of lobate livers.     

Iron derivatives from casein hydrolysates as a potential source in the treatment of iron deficiency

The properties of an Fe(3+)-peptide complex containing 5.6% Fe, obtained by the reaction of ferric chloride with an enzymatic hydrolysate of casein, are described. The major site of iron binding corresponds primarily to the carboxylate groups and to a lesser extent to the peptide bonds. The Fe(3+)-peptide complex is insoluble at acid pH and completely soluble at neutral to alkaline pH. When soluble, the Fe(3+) is tightly bound to the complex peptide mixture but can be displaced and complexed by a low molecular weight ligand such as cysteine. Its efficacy in relation to iron sulfate was compared in rats. Both iron sources were administrated in Milli-Q water by gastric gavage to male Wistar rats (180-200 g) after an 18 h fast with water ad libitum. Fe(3+) from the Fe(3+)-peptide complex was transferred to the blood in a dose-dependent manner (1-8 mg of Fe/kg), and the serum iron levels were significantly higher (p < 0.001) than in a similar group of rats treated with iron sulfate. In the comparative kinetics experiments, the rats received 4 mg of Fe/kg. Both iron sources presented maximum absorption, as indicated by the elevation of serum iron levels, 30 min after administration, and the AUC(0)(-->2h) of the Fe(3+)-peptide complex was significantly higher (p < 0.05) than that observed with iron sulfate. The simultaneous administration of free peptides (0-192 mg) with the Fe(3+)-peptide complex or iron sulfate did not modify the extent of absorption of iron from both sources, suggesting that the absorption is due to the complex formed and probably not to exchange reactions in the gastrointestinal tract. In the hemoglobin repletion experiments carried out on newly weaned rats with anemia induced by a low-iron diet, supplementation of the diet with the the Fe(3+)-peptide complex was as efficient as supplementation with iron sulfate in the conversion from diet to hemoglobin iron. These results, taken together, suggest that the Fe(3+)-peptide complex is a potential compound for use as an iron source in biological situations.     

Feasibility of automatic detection of knots in maritime pine timber by acousto-ultrasonic scanning

In order to assess the lengthwise variation of mechanical properties of maritime pine (Pinus pinaster Ait.) a study on the possibility of automatically detecting knots by ultrasonic readings across the grain was carried out. A through-transmission acousto-ultrasonic (AU) setup was implemented allowing acquisition of AU profiles along the central mid-third of each specimen. An algorithm was proposed and successfully implemented for enhancing variations on the AU profiles due to knot presence. Discriminant analysis was applied to the AU profiles. The results show the possibility of automatically differentiating clear wood zones from zones with a knot area ratio value above 20% or 30%, with a success rate of 75% to 83% depending on the dimension of the specimens tested.This research project was supported by the Foundation for Science and Technology (FCT) under contract Nos. PBIC/C/AGR/2296/95 and POCTI/33967/AGR/2000     

Photodynamic therapy for pythiosis

Background –  Pythiosis is a life‐threatening disease caused by Pythium insidiosum. Photodynamic therapy (PDT) is an alternative treatment to surgery that uses the interaction of a photosensitizer, light and molecular oxygen to cause cell death. Objectives –  To evaluate the effect of PDT on the in vitro growth of P. insidiosum and in an in vivo model of pythiosis. Methods –  For in vitro studies, two photosensitizers were evaluated: a haematoporphyrin derivative (Photogem^®) and a chlorine (Photodithazine^®). Amphotericin B was also evaluated, and the control group was treated with sterile saline solution. All experiments (PDT, porphyrin, chlorine and light alone, amphotericin B and saline solution) were performed as five replicates. For in vivo studies, six rabbits were inoculated with 20,000 zoospores of P. insidiosum, and an area of 1 cm³ was treated using the same sensitizers. The PDT irradiation was performed using a laser emitting at 660 nm and a fluence of 200 J/cm². Rabbits were clinically evaluated daily and histopathological analysis was performed 72 h after PDT. Results –  For in vitro assays, inhibition rates for PDT ranged from 60 to 100% and showed better results in comparison to amphotericin B. For the in vivo assays, after PDT, histological analysis of lesions showed a lack of infection up to 1 cm in depth. Conclusions and clinical importance –  In vitro and in vivo studies showed that PDT was effective in the inactivation of P. insidiosum and may represent a new approach to treating pythiosis.     

Growth, feed intake, carcass characteristics, and eating behavior of feedlot lambs fed high-concentrate diets containing soybean hulls

The objectives of this experiment were to determine the effects of replacing ground corn with soybean hulls (SH) in high-concentrate diets on the growth (56-d period), carcass characteristics, and eating behavior of feedlot lambs. Sixty-four Santa Inês ram lambs (18.3 ± 2.8 kg of BW and 69 ± 5 d of age) were assigned to a randomized complete block design experiment with 8 blocks and 4 diets. The control diet contained 10% coastcross (Cynodon sp.) hay, 70% corn, and no SH (SH0) in the dietary DM. In the remaining diets, SH replaced corn at the rate of 15 (SH15), 30 (SH30), or 45% (SH45) of the original corn concentration, resulting in 0, 10.5, 21.0, or 31.4% SH in the dietary DM. Dry matter intake increased linearly (P < 0.01) when SH replaced ground corn (1.0, 1.0, 1.1, and 1.1 kg/d for SH0, SH15, SH30, and SH45, respectively). There was no effect on ADG of lambs, with values of 276, 278, 282, and 287 g for SH0, SH15, SH30, and SH45, respectively. Feed efficiency decreased linearly (P < 0.01) with SH inclusion. Carcass measures were not affected by SH as a replacement for ground corn. Eating time, expressed as minutes per day and minutes per gram of NDF, showed a quadratic effect (P < 0.05), whereas no effect was observed when expressed as minutes per gram of DM. Rumination, in minutes per day, was not influenced by dietary SH inclusion, but a linear decrease (P < 0.01) was observed when this variable was expressed as minutes per gram of NDF. Soybean hulls can replace up to 45% of the ground corn (31.4% of SH in the dietary DM) in high-concentrate diets fed to feedlot lambs without negative effects on ADG and carcass measures. The linear decrease in feed efficiency (11.6% reduction from SH0 to SH45) suggests that optimal dietary SH inclusion rates should be dictated by the relative costs of SH and corn.     

Differential preference of <Emphasis Type="Italic">Capsicum</Emphasis> spp. cultivars by <Emphasis Type="Italic">Aphis gossypii</Emphasis> is conferred by variation in volatile semiochemistry

The aim of this study was to demonstrate that Capsicum spp. cultivars are differentially preferred by the cotton aphid, Aphis gossypii, and to investigate the role of volatile semiochemicals in conferring differences in host preferences. Two preference assays were conducted in 2008 under greenhouse conditions. Fourteen different commercially available cultivars were grown in cages protected by an anti-aphid net, and were infested 60 days after planting, through the release of ten adult female A. gossypii per plant. The results showed that after a five-day infestation period, statistically significant differences in the mean number of A. gossypii between cultivars were observed, with Sweet Pepper Hybrid Green Belt (SPHGB) being one of the cultivars with the lowest number of A. gossypii per plant. To test the hypothesis that the preference of cultivars was associated with release of volatile, Capsicum spp-derived semiochemicals, olfactometer behavior bioassays were conducted with A. gossypii, using volatile organic compounds (VOCs) collected from non-preferred SPHGB and preferred SPAB cultivars. A. gossypii was significantly repelled only by the VOCs of infested SPHGB. Furthermore, coupled gas chromatography-mass spectrometry (GC-MS) analysis of VOCs released by plants prior to, and after, A. gossypii infestation, revealed that the non-preferred SPHGB cultivar released nine additional compounds after infestation, including 6-methyl-5-hepten-2-one, a known plant defense semiochemical involved in plant—aphid interactions. These data suggest that non-preferred cultivars releasing this semiochemical have the potential to be used in breeding programs aimed at producing A. gossypii-resistant Capsicum spp. cultivars.     

Helical structures of ESCRT-III are disassembled by VPS4

During intracellular membrane trafficking and remodeling, protein complexes known as the ESCRTs (endosomal sorting complexes required for transport) interact with membranes and are required for budding processes directed away from the cytosol, including the budding of intralumenal vesicles to form multivesicular bodies; for the budding of some enveloped viruses; and for daughter cell scission in cytokinesis. We found that the ESCRT-III proteins CHMP2A and CHMP3 (charged multivesicular body proteins 2A and 3) could assemble in vitro into helical tubular structures that expose their membrane interaction sites on the outside of the tubule, whereas the AAA-type adenosine triphosphatase VPS4 could bind on the inside of the tubule and disassemble the tubes upon adenosine triphosphate hydrolysis. CHMP2A and CHMP3 copolymerized in solution, and their membrane targeting was cooperatively enhanced on planar lipid bilayers. Such helical CHMP structures could thus assemble within the neck of an inwardly budding vesicle, catalyzing late steps in budding under the control of VPS4.     

Resistance of Santa Ines and crossbred ewes to naturally acquired gastrointestinal nematode infections

This trial was carried out in Piracicaba, São Paulo State, Brazil, to comparatively evaluate the degree of resistance to naturally acquired gastrointestinal nematode infections in sheep of the following genetic groups: purebred Santa Ines (SI), SI crossbred with Dorper (DO × SI), Ile de France (IF × SI), Suffolk (SU × SI), and Texel (TE × SI). Fifteen ewes from each group were raised indoors until 12 months of age. At this age, they were moved to pasture that was naturally contaminated by nematode infective larvae and were evaluated from December to May, 2007. Rainfall ranged from 267 mm in January to 37 mm in April. Maximum and minimum mean temperatures ranged from 32.5 °C to 19.0 °C in March and from 25.9 °C to 12.8 °C in May. There was an increase in the mean number of eggs per gram of feces (EPG) after animals were placed on pasture with significant difference between the SI (80 EPG) and IF × SI (347 EPG) groups in January; and the DO × SI (386 EPG) and TE × SI (258 EPG) groups in May. The highest mean fecal egg count (FEC), 2073 EPG, was recorded for the TE × SI group in February. All groups showed a progressive reduction in body weight throughout the experiment of 12.0% (TE × SI) to 15.9% (SU × SI). In general, the animals with the highest FEC presented the lowest packed cell volumes (PCV); the highest correlation coefficient between FEC × PCV occurred in the SU × SI sheep in January (r = −0.70; P < 0.01). Similarly, there was an inverse relationship between FEC and blood eosinophil values, with the highest correlation coefficient in the TE × SI sheep in February (r = −0.64; P < 0.05). Immunoglobulin G (IgG) levels against Haemonchus contortus antigens increased in all groups as a result of the exposure to parasites and remained relatively constant until the end of the study, with the exceptions of SU × SI and TE × SI, which showed a rise in IgG levels during the last sampling that coincided with a reduction in mean FEC. In conclusion, crossbreeding Santa Ines sheep with any of the breeds evaluated can result in a production increase and the maintenance of a satisfactory degree of infection resistance, especially against H. contortus and Trichostrongylus colubriformis, the major nematodes detected in this flock.