Polymorphism of P53‐Ets/AP1 transactivation region of MDM2 oncogene and its immunohistochemical analysis in canine tumours

Mouse Double Minute‐2 (MDM2) is an ubiquitin ligase which is overexpressed or its promoter polymorphism has been reported in different tumours. The objective of this study was to examine the MDM2 protein expression and its promoter polymorphism in some canine tumours. Twenty specimens were collected from 20 dogs with 15 mammary gland carcinomas, 3 lymphomas, 1 transmissible venereal tumour and 1 trichoblastoma. Samples were analysed immunohistochemically using human antibody against MDM2 protein. PCR and DNA sequencing were carried out to identify MDM2 promoter polymorphism. MDM2 gene was expressed in 13 of 20 samples including 11 mammary carcinomas, 1 lymphoma and 1 trichoblastoma. We found 94% homology between canine and human sequences. Four mutations including G169C, A177G, G291T and A177G were identified in different types of breast carcinomas. An extra p53 response element was found in a mixed mammary carcinoma.     

Influence of storage and drying methods on invertebrate elemental and isotopic measurements

Optimised pre-analytical methods for measuring the chemical properties of soil macro-organisms are needed. We tested the effects of ethanol immersion, freezer storage, and drying method on the reliability of estimates of key stoichiometric elements (carbon [C], nitrogen [N], phosphorus [P]) and abundances of ¹³C and ¹⁵N in samples of crickets (Acheta domestica), cockroaches (Nauphoeta cinerea) and mealworms (larval Tenebrio molitor). Ethanol immersion tended to increase A. domestica C and N, and reduced A. domestica P, relative to the reference treatment (deep freezing and oven drying). For N and P these effects were only present after 28-day ethanol immersion. Nauphoeta cinerea and T. molitor samples were generally unaffected by storage treatments, while δ¹³C and δ¹⁵N were not affected by any storage treatment for any species. Thus, five days of ethanol immersion may be acceptable prior to elemental and stoichiometric analyses of hard-bodied soil invertebrates in comparative studies.     

Dietary simultaneous replacement of fish meal and fish oil with blends of plant proteins and vegetable oils in yellowfin seabream (Acanthopagrus latus) fry: Growth,digestive enzymes,antioxidant status and skin mucosal immunity

A 56‐day nutritional research was performed to examine the influence of alternative vegetal protein and lipid sources on performance of yellowfin seabream fry (Acanthopagrus latus) (0.5 ± 0.0 g). In this regard, five isoproteic (Ca. 500 g/kg) and isolipidic (Ca. 150 g/kg) diets were formulated in which fish meal (FM) and fish oil (FO) were simultaneously replaced with blends of plant proteins (PP, soybean meal and corn gluten) and vegetal oils (VO, canola and soybean oils) at 20% (SR20), 40% (SR40), 60% (SR40) and 80% (SR80) levels, respectively; meanwhile, a control diet (SR0) was formulated based on FM and FO. Growth and feed utilization were not influenced by experimental diets. The fatty acid profile of fillet drastically altered by dietary treatments. Fish fed with the SR60 and SR80 feeds had higher total protease, trypsin and α‐amylase activities than other treatments. The antioxidant enzyme activities and glutathione content in liver were enhanced in fish fed with the SR40, SR60 and SR80 diets. Skin mucosal immune parameters including total protein content, alkaline phosphatase and alternative complement pathway activities in the control group were relatively lower than the vegetal treatments. According to these results, it is recommended that 410 g/kg of FM and 45 g of FO/kg can be replaced with alternative vegetal sources in diet for A. latus fry.     

Extrapituitary gonadotropin-releasing hormone (GnRH) binding sites in goldfish

In teleosts, as in other vertebrates, the secretion of pituitary gonadotropin (GTH) is mediated by the hypothalamic decapeptide, gonadotropin-releasing hormone (GnRH). Recent findings in teleosts indicate that GnRH receptors are not restricted to the pituitary gonadotropes and are also associated with somatotropes as well as being present in a number of other tissues. In the present study, we provide novel information on GnRH binding in a number of extrapituitary tissues in goldfish. However, we do not intend to provide full characterization of GnRH binding sites in various extrapituitary tissues in goldfish as this would clearly be outside the scope of this paper. In this study we examined GnRH binding in a number of extrapituitary tissues in goldfish and observed specific binding in ovary, testis, brain, liver and kidney. No specific GnRH binding was observed in muscle, skin, gut, gill and heart. In general, the present findings together with the results of other studies carried out in our laboratory demonstrate that mature goldfish ovary and testis contain two classes of GnRH binding sites, high affinity/low capacity and low affinity/high capacity sites with binding characteristics similar to those of the pituitary GnRH receptors. The brain of goldfish was also found to contain two classes of GnRH binding sites, a super-high affinity/low capacity and a low affinity/high capacity sites. Furthermore, study of goldfish liver and kidney demonstrated the presence of a single class of GnRH binding sites with characteristics different from those of pituitary, ovary, testis and brain. Overall, it is evident that goldfish contains a family of GnRH binding sites which can be classified into four groups based on binding affinities: 1) A class of high affinity binding sites present in the pituitary, ovary and testis, 2) a class of super high affinity sites so far only detected in the brain, 3) a class of intermediate-affinity GnRH binding sites in the liver and kidney, and 4) a class of low affinity binding sites present in all the tissues containing specific GnRH binding sites except for liver and kidney.     

A study of goldfish oocyte meiosisin vitro: effects of 2,4-dinitrophenol and adenosine-5-triphosphate

Germinal vesicle migration (GVM) and/or dissolution (GVD) were measured in goldfish oocytes, treated with 17α, 20β dihydroxyprogesterone (DHP) and other compounds considered to effect the cytoskeleton and oxidative phosphorylation,in vitro. Administration of DHP reinitiated meiotic maturation, increasing GVM and GVD in goldfish oocytes. Addition of 2,4-dinitrophenol (DNP) to the incubation medium significantly inhibited DHP-induced GVM and GVD. The DNP effect was found to be partially reversible after 24 h and could be reversed fully after a further delay of approximately 24h. Treatment of goldfish oocytes with demecolcine (DE; a colchicine derivative also known as colcemid) induces GVM to the micropyle without effecting GVD; while Cytochalasin-B which inhibits microfilament polymerization impairs both GVM and GVD. Administration of DNP, significantly inhibited DE-induced GVM, suggesting that GVM as well as GVD are dependent upon the process of oxidative phosphorylation. Addition of adenosine-5′ -triphosphate (ATP) at low concentrations (0.01–0.1 mM) did not effect DHP-induced or DNP-inhibited GVD in goldfish oocytes. The present results are consistent with the idea that migration of the oocyte nucleus during meiosis reinitiation has an energy requirement and involves participation by the cytoskeleton.     

Surgical treatment of a mammary gland comedocarcinoma in an Arabian mare: Post-operative management,and histopathological and immunohistochemical features

A lactating 20-year-old, brown, Arabian mare, weighing about 300 kg, presented for bleeding from one teat and severe swelling of the entire mammary gland. The mare had untreated mastitis 10 months before. Consequently, a gangrenous teat developed after chronic bloody and purulent discharges. The teat was removed surgically by the field veterinarian. At that time, the mammary gland increased in size. Bloody and purulent discharges restarted 10 days previously. Under general anaesthesia, the entire mammary gland was removed. Comedocarcinoma was diagnosed by histopathological assessment. Immunohistochemical staining was performed for pan-cytokeratin and vimentin. Microscopic examination of immunohistochemical stained slides revealed expression of pan-cytokeratin. In conclusion, this report describes clinical, macroscopic, histopathological and immunohistochemical characteristics of comedocarcinoma that did not metastasise to regional lymph nodes. Reports in the field of equine oncology contribute to improved general knowledge in equine medicine, contributing to better diagnosis and treatment.     

In ovo injection of flavanone on bone quality characteristics,biochemical parameters and antioxidant enzyme status of blood in daily chicks

In ovo injection (IOI) of Naringin (N), flavanone was examined on post‐hatch blood biochemical parameters, antioxidant status and bone characteristics. Fertile eggs (n = 700) were distributed in seven groups with 100 eggs. On 14th and 17.5th days of incubation, four groups were injected using 15 or 30 mg active ingredient levels of naringin/0.5 ml saline/egg, low and high level, into amnion sac. Controls include sham (injected normal saline, 0.5 ml/egg on day 14 and 17.5th) and un‐injected group. IOI of high naringin and saline on 14th day of incubation resulted in lower hatchability and then higher mortality in last week of embryonic life. On day hatch, high levels of injected groups more body weight compared to the control. Chick length was increased at high levels of naringin on day 17.5th compared to control and saline injected. Quality traits of bones were improved in naringin‐injected groups compared to control. IOI of naringin influenced thyroid hormones on 14th day of incubation. Naringin groups influenced the Alkaline phosphatase (ALP), Calcium (Ca), superoxide dismutase (SOD), blood biochemical and lipids. Totally, amniotic IOI of naringin in last days of developing embryo may be useful for hatched chick, development of leg long bone or effect on biochemical metabolites by levels of flavanone that it needs more research.     

The distribution and frequency of damage to roadside trees in low-volume road construction

We investigated the distribution and frequency of damage to tree stands adjacent to low-volume roads according to the type of hillside materials involved(soil or rock) and hillside gradient in mountainous forests of northern Iran. A total of 80 plots were systematically and randomly sampled to record damaged trees(bending,crushing and wounding) by class of hillside gradient and materials at the edge of road. Tree wounding and crushing at rock slopes was significantly greater than at hillsides with a mix of clay soil(p \ 0.05). Damage on hillsides with slope gradients[45% were 2, 8.5 and 2.3 times more frequent than on hillsides with slope gradient\15% for bending, crushing and wounding, respectively. The damage distribution varied according by type and the most frequent damage was tree wounding(p \ 0.05). The damage distribution was measured at distances of 4, 5 and 8 m from the road fillslope for tree bending, crushing and wounding, respectively. Using hydraulic excavators and physical barriers(wooden obstruction and synthetic holder) during earthworks for road construction could reduce these damage.     

Iron Fortification of Whole Potato Using Vacuum Impregnation Technique with a Pulsed Electric Field Pretreatment

In this study, the effect of a vacuum impregnation (VI) process with pulsed electric field (PEF) pretreatment on fortification of whole potato with ferric pyrophosphate was investigated. The VI process and PEF pretreatment were optimized with respect to physicochemical attributes of PEF-VI-processed samples using response surface methodology (RSM). The results indicated that the iron content of VI-processed and PEF-VI-processed potatoes was increased about 126 and 457%, respectively, compared to unprocessed potatoes. A more intensive PEF pretreatment led to a higher cell disintegration, decrease in colour and textural parameters and also increase in components leakage from treated potato. Our study suggests that whole 180-g potatoes should be pretreated at 394 V/cm with 36 pulses and then treated at 3.5 kPa for 37 min in vacuum stage and 39 min restoration time in atmospheric stage to achieve the highest iron fortification and the lowest physicochemical changes during PEF-VI processing.