Effects of dietary fish to rapeseed oil ratio on steatosis symptoms in Atlantic salmon (Salmo salar L) of different sizes.

Choline is recognized as an essential nutrient for Atlantic salmon at all developmental stages. However, its dietary requirement is not well defined. Choline plays a critical role in lipid transport, and the clearest deficiency sign is intestinal steatosis. The present work, aiming to find whether lipid source and fish size may affect steatosis symptoms, was one of a series of studies conducted to identify which production-related conditions may influence choline requirement. Six choline-deficient diets were formulated varying in ratios of rapeseed oil to fish oil and fed to Atlantic salmon of 1.5 and 4.5 kg. After eight weeks, somatic characteristics were observed, and the severity of intestinal steatosis was assessed by histological, biochemical, and molecular analyses. Fatty acid composition in pyloric intestine, mesenteric tissue, and liver samples was also quantified. The increasing rapeseed oil level increased lipid digestibility markedly, enhancing lipid supply to the fish. Moreover, small fish consumed more feed, and consequently had a higher lipid intake. In conclusion, the results showed that choline requirement depends on dietary lipid load, which depends on the fatty acid profile as well as the fish size.

Impact of different methodologies on the detection of point mutations in routine air-dried fine needle aspiration (FNA) smears.

Currently the best method to select suspicious thyroid nodules for surgery is fine needle aspiration (FNA) cytology. However, FNA cytology has some inherent limitations, which can partly be overcome by molecular analysis. Therefore, molecular testing for somatic mutations has emerged as the most promising approach for molecular FNA diagnostics. The objective of this methodological study was to evaluate the feasibility of detecting BRAF, NRAS, HRAS, and KRAS mutations from routine air-dried thyroid FNA smears, and to find an optimal method for detecting these mutations in FNA samples. DNA was extracted from 110 routine air-dried FNA smears and the corresponding surgically obtained formalin-fixed paraffin-embedded tissues. The presence of BRAF, NRAS, HRAS, and KRAS mutations was assessed by real-time PCRs and high resolution melting analysis, and/or pyrosequencing in comparison to real-time PCRs using hybridization probes and fluorescence melting curve analysis. The high-resolution melting-PCRs revealed a significantly lower number of PCR failures and questionable results, and detected more mutations than the PCRs using hybridization probes. The number of PCR failures ranging from 14-16% by high-resolution melting-PCRs could be further reduced to 5-14% by adding pyrosequencing assays. Moreover, pyrosequencing increased the specificity of the assays, up to 98-100%, while the sensitivity ranged between 32-63%. In summary, the mutation detection, especially in air-dried FNA samples, improves when using PCR assays in combination with high resolution melting analysis. Additional improvement can be obtained by subsequent pyrosequencing in comparison to previously described real-time PCRs using hybridization probes and fluorescence melting curve analysis.

The prognostic significance of histological features in oral squamous cell carcinoma.

BACKGROUND: Different factors predict nodal metastasis, recurrence and survival in oral cancer. The aim was to assess the prognostic value of histological features related to the primary tumour. METHODS: A total of 144 patients surgically treated at Odense University Hospital for oral cancer between 1999 and 2004 were included in the study. Postoperative radiation therapy was given in case of close and involved margins or high TNM stages (UICC 1997). Median follow-up time was 38 months. All surgical resections were reviewed and 22 histological characteristics were assessed. RESULTS: The predominant sites were floor of mouth (FOM, 39%) and lateral tongue (35%). Fifty-nine per cent had UICC97 stage I-II disease. Five-year cause-specific survival was observed in 65%. Nodal involvement at diagnosis was observed in 36% which was significantly related to grade, neural and vascular invasion; surgical margins and increasing tumour depth. A cut-off value of 2 mm (4 mm for FOM) separated patients without and with nodal metastasis at the time of diagnosis. However, on multivariate analysis, neck disease was only associated with tumour depth and grade. Cox analysis of local recurrence in the oral cavity over time showed that tumour diameter and surgical margins were significant predictors while cause-specific survival was related to diameter, depth of invasion, surgical margins and extracapsular spread (ECS). CONCLUSIONS: Tumour depth and grade were strong prognostic factors for nodal metastasis, independently of other histological features. Tumour diameter and margins independently predict local recurrences in the oral cavity as well as cause-specific survival. Nodal involvement and ECS were associated with adverse prognosis.

Atlantic salmon (Salmo salar L.) parr fed genetically modified soybeans and maize: Histological, digestive, metabolic, and immunological investigations.

Physiological and health related responses to dietary inclusion of genetically modified (GM) full-fat soybean meal (Roundup Ready; GM-soy) and maize (MON810 Bt-maize; GM-maize), as well as non-parental, untransformed lines (nGM-soy and nGM-maize D2), were evaluated in farmed Atlantic salmon (Salmo salar L.) parr during the first 8 months of feeding. Significant effects of dietary GM presence were only found in intestinal Na+-dependent d-glucose uptake and SGLT1 protein level in the region pyloric caeca in which the highest values were found in the GM-soy, intermediate in the nGM-soy, and lowest in the standard FM fed groups. Data from this study confirm that GM soybeans (RRS) and maize (MON810) at inclusion levels of about 6% appear to be as safe as commercially available nGM soy and maize in diets for Atlantic salmon parr. Results from studies with higher inclusion levels and with non-modified, isogenic or near-isogenic parental lines as control groups are pending.

Laryngeal cancer and human papillomavirus: HPV is absent in the majority of laryngeal carcinomas.

Thirty laryngeal carcinomas from patients without pre-existing laryngeal papillomatosis were examined by PCR for the presence of HPV DNA. The utmost care was taken during sectioning of the tissue blocks and DNA-extraction in order to avoid false positive results. Three pairs of consensus primers were used: MY9/MY11, GP5+/GP6+ and CPI/CPII. HPV was detected in 1/30 carcinomas. The HPV type present could not be determined, but it was not type 6, 11, 13, 16, 18, 30, 31, 33, 35 or 45. In other studies the reported frequency of HPV in laryngeal carcinomas, as estimated by PCR, varies between 3-85%. The reasons for this unacceptable variation in reported results are discussed. The present results indicate that HPV DNA does not have a major role in malignant tumours of the larynx in patients without pre-existing recurrent laryngeal papillomatosis.

A case of granulomatous sialadenitis of the submandibular gland.

Granulomatous inflammation of the major salivary glands is very rare and may be due to obstruction. Little attention has been paid to this condition. The reaction is caused by extravasation of mucus, as seen in the common mucocele of the minor salivary glands. A case of granulomatous inflammation of the submandibular gland caused by obstruction is presented. The etiology of granulomatous sialadenitis is reviewed.

Recurrent pregnancy-related upper airway obstruction caused by intratracheal ectopic thyroid tissue.

An unusual case of recurrent pregnancy-related thyroid growth stimulation is reported. A 27-year-old euthyroid woman had pulmonary symptoms, thought to be asthma during her first pregnancy, that improved postpartum. Bronchodilatators had no effect and symptoms recurred from gestational week 22 during her second pregnancy. Her 58-mL multinodular goiter (by ultrasound) was not thought to be responsible for her upper airway symptoms. Therefore, fiber laryngoscopy and computed tomographic (CT) scan were performed and revealed a 20 x 15 x 10 mm intratracheal tumor. After tracheostomy and microlaryngoscopy, benign goitrous thyroid tissue was removed through a tracheal fissure during gestational week 35. Postoperatively the patient had stopped medication and was without any pulmonary symptoms. The child was delivered by cesarean section in gestational week 39. Apgar score was normal and the child has developed normally. We believe that this case illustrates the recurrent effect of pregnancy-related thyroid tissue stimulation by a combination of increasing human chorionic gonadotropin (hCG) stimulation and iodine deficiency in a borderline iodine-deficient region. This is the first report on symptomatic intratracheal ectopic thyroid tissue diagnosed during pregnancy.

Effects of soybean meal and salinity on intestinal transport of nutrients in Atlantic salmon (Salmo salar L.) and rainbow trout (Oncorhynchus mykiss).

Groups of fresh- and seawater-adapted Atlantic salmon (Salmo salar L.) and rainbow trout (Oncorhynchus mykiss) were fed diets with (SBM diet) or without (control diet) extracted soybean meal (30% of protein substituted with SBM) for 3 weeks. Average fish size per group ranged from 597 to 1763 g. One tank or net pen per species, dietary group and water salinity was used. In vitro nutrient transport (D-glucose, the L-amino acids aspartate, lysine, methionine, phenylalanine and proline, and the dipeptide glycyl-sarcosine) was measured using intact tissue (everted sleeve method) from the different postgastric intestinal regions. The dimensions of the different intestinal regions were also measured for each treatment group. Results indicate that SBM causes decreased carrier-mediated transport and increased permeability of distal intestinal epithelium for the nutrients, and the capacity of this region to absorb nutrient was diminished. Salinity may also affect the relative contribution of carrier-mediated and independent uptake to total nutrient absorption.

A glucokinase-like-enzyme in the liver of Atlantic salmon (Salmo salar).

An enzyme with properties similar to rat liver glucokinase (Hexokinase IV or D) is present in salmon liver in addition to low-Km hexokinase(s). The specific activity of this enzyme increases about 1.6 fold, comparing activities after feeding diets with 25% and 0% digestive energy from starch. The enzyme has a low affinity for glucose, S0.5 = 25.2-26.8 mM (95% confidence interval) and a low activity with fructose, approximately 8% of the activity with glucose. Its molecular mass was estimated to 50.7 +/- 0.6 kDa (SEM. n = 3) by gel filtration, and it displays positive cooperativity with respect to glucose. The Hill constant = 1.73-1.81 (95% confidence interval). The enzyme is competitively inhibited by N-acetyl glucosamine, K(i) approximately 0.28 mM.

Carcinoma occurring in branchial cleft cysts.

In order to find histological data in the differentiation between branchial cleft carcinomas and metastatic carcinomas, the specimens from 154 patients with branchial cleft cysts and 7 patients with an isolated tumour in the neck with unknown primary tumour were reviewed and compared with 10 normal lymph nodes. Absence of lymph node structures as peripheral lobulation, internodular trabeculae and perinodular sinuses in branchial cleft cystc, are found valuable for distinguishing primary carcinoma of branchial cleft cysts from metastases. A correct diagnosis of this rare tumour is important in order to avoid overtreatment of these patients, who have a good prognosis if treated with surgical excision only.

Spindle cell carcinoma of the larynx.

Histology, histogenesis, biology and treatment of the rare spindle cell carcinoma of the larynx are presented. The histogenesis of spindle cell tumours has been much debated, but electron microscopic investigations have shown that most of the laryngeal lesions are squamous epithelial carcinomas that have undergone transformation to a spindle cell pattern. As confusion with supporting tissue neoplasms may occur in the light microscope, the use of the electron microscope in differential diagnostics is recommended.

Estimation of nitrogen digestibility in poultry: content and distribution of major urinary nitrogen compounds in excreta.

The investigation concerned the digestibility of nitrogen and the content of uric acid, ammonia, and urea in urine and feces in White Leghorn layers. Eight colostomized hens were given a commercial cage-layer diet. Feces and urine were collected under acid conditions. Acidified urine contained more ammonia than untreated urine. For quantitative determination of ammonia, Urine collection ought to be done under acid conditions. Of urinary nitrogen uric acid represented 88%, ammonia 7%, urea 3%, and 2% unidentified. Urinary ammonia constituted 90% of ammonia in total excreta and urinary urea 86% of the urea in excreta. By means of these distribution factors and content of nitrogen compounds in total excreta, good estimates of the nitrogen digestibility were obtained. Our method appears to be applicable to chickens of other ages, sex, breeds, and environmental conditions.

Influence of age on lipase, amylase, and protease activities in pancreatic tissue and intestinal contents of young turkeys.

Day-old male turkeys were fed either a reference diet with 12% sucrose or experimental diets with 12% tallow or 12% animal-vegetable blend (A-V fat) replacing sucrose until 56 days of age. Poults were sampled at 1 day of age and every 2 to 7 days thereafter for determination of enzyme activities of pancreas and contents of the proximal one-fourth of the small intestine. In pancreatic tissue, trypsin, protease, and lipase activities increased with age after a lag period of about 14 days. Amylase activity increased rapidly during the first 14 days. In intestinal contents, trypsin, protease, and amylase increased from Day 1 until Day 21. Development of intestinal lipase activity seemed to depend on dietary fat level. Low activities were observed with low fat diets throughout the study. With high fat diets, a lag period of about 3 wk was followed by a five-fold increase in lipase activity.

Influence of age on utilization of supplemental fats by young turkeys.

Male poults were fed diets containing 0, 4, 8, or 12% tallow or 4, 8, or 12% animal-vegetable fat blend (A-V Fat) from 1 day to 8 weeks of age. Supplemental fat improved 8-week body weight and feed efficiency, irrespective of fat source. Fat retention (fat consumed-fat excreted) was not affected by fat level or by fat source after 2 weeks of age. At 2 weeks, fat retention by poults fed tallow was less than that of poults fed A-V Fat. Fat retention increased with age, irrespective of fat source or level. Fat retentions ranged from 66.4 to 83.7 at 2 weeks and from 90.8 to 96.5 at 8 weeks of age. Changes in metabolizable energy (ME) values of the fats with age paralleled those of fat retention, whether ME were estimated by multiplying fat retention percentages times gross energy of the fats or by regression analysis of changes in diet ME associated with increments of fats. On the basis of changes in diet ME, the ME of tallow was 6808 kcal/kg at 2 weeks and 8551 kcal/kg at 8 weeks. Analagous values for A-V Fat were 7114 and 8924 kcal/kg, respectively. Fatty acid analysis of dietary and excreta fat indicated that, as turkeys aged, improvements in retention were greatest for palmitic and stearic acids, especially in the instance of tallow. Fatty acid binding protein (FABP) assay of intestines from poults fed no added fat, 12% tallow, or 12% A-V Fat showed that fat level did not significantly affect FABP activity. There was, however, a marked increase in intestinal FABP in 6- and 8-week-old poults as compared with 2- or 4-week-old poults.

Plasma cell gingivitis apparently related to the use of khat: report of a case.

Plasma cell gingivitis (PCG) is characterized by massive infiltration of plasma cells into the subepithelial tissue. It is a rare condition; the cause of which is still not fully understood. A case of PCG is reported in the mandibular gingiva probably caused by chewing khat. This report is the first, as far as we know, that relates PCG to the use of khat. The histological examination revealed infiltration of polyclonal plasma cells without signs of fungus, tuberculosis or malignancy. It is concluded that the changes were compatible with an allergic-like reaction. The patient, a 30-year-old immigrant from Somalia, revealed in a subsequent consultation that he regularly used khat. The leaves are placed in the buccal sulcus. The PCG disappeared within two weeks of the use of khat being discontinued. Dental surgeons (periodontists) in Europe and the New World will, due to increasing immigration from Africa and the Arabian Peninsula, meet more patients who regularly use khat. This means that PCG and other khat related intraoral changes will become more common in the future.

Oncocytomas of the salivary glands.

Two cases of oncocytoma of the parotid gland are presented, one of which was dominantly cystic and had recurred after 21 years. The light and electron, microscopic pictures are presented. The literature has been reviewed, and 131 cases were found in salivary glands; hence, oncocytomas in this location are rare. In older persons, they are found predominantly in the parotid gland, but have also been reported in the minor salivary glands. It is pointed out that electron microscopy or histochemical examination is indispensable for the demonstration of mitochondrial hyperplasia.

[Milker's nodules]. / Malkerknuder.

Milker's nodule is a parapox virus infection seen mostly on the hands of dairy farmers. We saw 15 cases over a period of two years in the County of North Jutland. Clinically, milker's nodule goes through a papular, a nodular and a crusted stage. Most patients were seen when the infection was in the nodular stage, an often painful condition requiring treatment. Three patients developed an erythema multiformelike secondary eruption. Lesions from nine patients were removed for histological examination. The histology of all lesions was consistent with milker's nodule. In three of seven patients parapox virus was demonstrated by electron microscopy. Treatment was commonly curettage followed by cauterization.

[Negative effects of anti-nutritional factors from soybeans in Salmonidae]. / Negatieve effecten van antinutritionele factoren in soyabonen bij salmoniden.

Soya bean products are used with increasing frequency in diets for salmonids. Soya beans however contain antinutritional factors (ANF), like protease inhibitors and lectins, which may decrease growth performance and even cause disease. The paper reviews the literature and adds some recent developments to the knowledge on the negative effects of ANF from soya beans in salmonids.

Context-dependent responses to novelty in Rainbow trout (Oncorhynchus mykiss), selected for high and low post-stress cortisol responsiveness.

Previous studies in a rainbow trout model, selectively bred for high (HR) and low (LR) post stress plasma cortisol levels, have yielded data that are indicative of contrasting stress coping styles. Fish from the HR line have been suggested to display a more diverse behavioral repertoire in challenging situations than the LR counterpart. The present study addressed whether such variation in behavioral flexibility traits was evident in different experimental settings using these selection lines. The fish were subjected to three sets of challenges (novel object test, resident-intruder test and confinement stressor test), all which were repeated a week later. Introducing a novel object evoked a divergent behavioral response in association with feeding: fish from the LR line displayed consistently suppressed feed intake while the HR fish remained unaffected. This observation was found to be repeatable along with attack latency and movement activity from the resident-intruder and confinement stressor tests. These results indicate that the behavioral responses in this animal model are context-dependent and shed new light on the expression of behavioral flexibility.