QTL and association analysis for skin and fibre pigmentation in sheep provides evidence of a major causative mutation and epistatic effects.

The pursuits of white features and white fleeces free of pigmented fibre have been important selection objectives for many sheep breeds. The cause and inheritance of non-white colour patterns in sheep has been studied since the early 19th century. Discovery of genetic causes, especially those which predispose pigmentation in white sheep, may lead to more accurate selection tools for improved apparel wool. This article describes an extended QTL study for 13 skin and fibre pigmentation traits in sheep. A total of 19 highly significant, 10 significant and seven suggestive QTL were identified in a QTL mapping experiment using an Awassi × Merino × Merino backcross sheep population. All QTL on chromosome 2 exceeded a LOD score of greater than 4 (range 4.4-30.1), giving very strong support for a major gene for pigmentation on this chromosome. Evidence of epistatic interactions was found for QTL for four traits on chromosomes 2 and 19. The ovine TYRP1 gene on OAR 2 was sequenced as a strong positional candidate gene. A highly significant association (P < 0.01) of grandparental haplotypes across nine segregating SNP/microsatellite markers including one non-synonymous SNP with pigmentation traits could be shown. Up to 47% of the observed variation in pigmentation was accounted for by models using TYRP1 haplotypes and 83% for models with interactions between two QTL probabilities, offering scope for marker-assisted selection for these traits.

Management of parapharyngeal space cavernous sinus haemangioma using transoral robotic surgery.

Cavernous haemangiomas are a very rare occurrence in the parapharyngeal space (PPS). Here, we present a case of a 58-year-old woman with an incidentally identified left PPS mass thought to be a pleomorphic adenoma that underwent excision by transoral robotic surgery (TORS). Intraoperative findings demonstrated no solid mass present and histological assessment of resected tissue confirmed a cavernous haemangioma. We discuss our experience in the management of a radiological and surgical mismatch of a PPS mass in what we believe to be the first cavernous haemangioma to be excised by TORS in this region.

Cigarette smoke inhibits macrophage sensing of Gram-negative bacteria and lipopolysaccharide: relative roles of nicotine and oxidant stress.

BACKGROUND AND PURPOSE: Smoking cigarettes is a major risk factor for the development of cardiovascular and respiratory disease. Moreover, smokers are more prone to infections. This has been associated with a suppression of the immune system by smoke. However, it is not clear how cigarette smoke affects the ability of immune cells to sense pathogens. Cigarette smoke contains a large number of molecules which may mediate responses on immune cells and of these, nicotine and oxidants have both been identified as inhibitory for the sensing of bacterial lipopolysaccharide (LPS). Nitric oxide synthase (NOS) and tumour necrosis factor (TNF)-alpha are both induced in macrophages on stimulation with Gram negative bacteria or LPS. EXPERIMENTAL APPROACH: We used murine macrophages stimulated with whole heat-killed bacteria or LPS. We measured output of NO (as nitrite) and TNFalpha, NOS protein by Western blotting and cellular oxidant stress. KEY RESULTS: Cigarette smoke extract suppressed the ability of murine macrophages to release NO, but not TNFalpha in response to whole bacteria. Cigarette smoke extract also inhibited nitric oxide synthase II protein expression in response to LPS. The effects of cigarette smoke extract on nitrite formation stimulated by LPS were unaffected by inhibition of nicotinic receptors with alpha-bungarotoxin (100 units ml(-1)). However, the effects of cigarette smoke extract on LPS-induced nitrite formation were mimicked by hydrogen peroxide and reversed by the anti-oxidants N-acetyl cysteine and glutathione. CONCLUSIONS AND IMPLICATIONS: We suggest that cigarette smoke exerts its immunosuppressive effects through an oxidant-dependent and not a nicotine-dependent mechanism.

Protein catabolism in advanced renal disease: role of cytokines.

BACKGROUND: Protein energy wasting is a maladaptive metabolic state often associated with inflammation, which is common in patients with chronic kidney disease (CKD). METHODS: A literature search was performed using MEDLINE and the reference lists of relevant review articles. The following key words were used in the MEDLINE search: "cytokines", "inflammation", "protein metabolism", "acute-phase protein", "cachexia", "chronic kidney disease", "end-stage renal disease" and "hemodialysis". The search was limited to English-language articles. RESULTS: While experimental models have shown that uremic animals are more prone for proteolysis, the results from the human studies are controversial. Intradialytic loss of amino acids and activation of proinflammatory cytokines lead to protein catabolism during hemodialysis (HD). At the whole-body level, intradialytic parenteral nutrition (IDPN) increases protein synthesis and decreases proteolysis. Amino acid infusion during HD increases muscle protein synthesis, but does not decrease protein catabolism. Activation of interleukin-6 during HD induces protein catabolism, impairs amino acid utilization for protein synthesis and increases acute-phase protein synthesis. CONCLUSION: The changes in albumin, fibrinogen and muscle protein kinetics during HD could be due to competing and complementary effects of availability of amino acids and activation of proinflammatory cytokines.

A genetic selection for isolating cDNAs encoding secreted proteins.

We describe a simple, rapid technique for simultaneously isolating large numbers of cDNAs encoding secreted proteins. The technique makes use of a facile genetic selection performed in a strain of Saccharomyces cerevisiae deleted for its endogenous invertase gene. A cDNA cloning vector which carries a modified invertase gene lacking its leader sequence is used in conjunction with this strain. Heterologous secreted genes fused appropriately upstream of this defective invertase provide the necessary signals to restore secretion, allowing the yeast to grow on sugars such as sucrose or raffinose. This microbial growth selection facilitates scanning cDNA libraries containing millions of clones, enabling the wholesale identification of novel secreted proteins without the need for specific bioassays. The technique is similar to one previously described (Klein et al. (1996) Proc. Natl. Acad. Sci. USA 93, 7108-7113). We describe results using a cDNA library derived from activated human peripheral blood mononuclear cells (PBMC). Genes identified from this library encoded signal sequences of proteins of diverse structure, function, and cellular location such as cytokines, type 1 and type 2 transmembrane proteins, and proteins found in intracellular organelles. In addition, a number of novel secreted proteins were identified, including a chemokine and a novel G-protein-coupled receptor. Since signal sequences possess features conserved throughout evolution, the procedure can be used to isolate genes encoding secreted proteins from both eukaryotes and prokaryotes.

Metal complexes of antiinflammatory drugs. Part VII: Salsalate complex of copper(II).

The preparation and properties of the Cu(II) complex Cu(SAS)2.H2O are reported for the antiinflammatory drug Salsalate (SAS). The diffuse reflectance spectra and magnetic moments are consistent with a dinuclear structure as found for [Cu(aspirinate)2(H2O)]2. The Cu(II) complex exhibits an increased superoxide dismutase activity compared with the parent drug molecule in the nitroblue tetrazolium assay.

Ultrasonic assessment of hemidiaphragmatic movement: an indirect method of evaluating mediastinal invasion in non-small cell lung cancer.

This preliminary study aimed to assess the potential value of diaphragmatic ultrasound (DUS) in evaluating phrenic nerve involvement indirectly, in the non-invasive pre-operative staging of mediastinal invasion in non-small cell lung cancer (NSCLC). A prospective study of 30 patients with NSCLC comparing the findings of diaphragmatic ultrasound, chest radiograph, computed tomography (CT) thorax and mediastinoscopy was performed. In all cases adequate quantitiative assessment of hemidiaphragmatic excursion was obtained. There was discordance in four of 30 patients between DUS and chest radiograph. Three of nine patients with extensive mediastinal disease on CT had abnormal DUS, and two of eight patients with indeterminate mediastinal disease on CT had abnormal DUS and were later found to be non-resectable. No abnormal cases of DUS were found in those cases with normal mediastinal CT. There was no clear relationship between the site, size and side of the primary tumour on CT, or its pleural or diaphragmatic contiguity, and hemidiaphragmatic excursion. There was concordance between DUS and mediastinoscopy in 17 of 21 patients. Two patients had normal mediastinoscopy and abnormal DUS but were not resectable at thoracotomy. No patient with abnormal DUS was resectable. DUS may be of potential value in the pre-operative staging of NSCLC and is therefore worthy of further evaluation.

Intestinal atresia with gastroschisis: a selective approach to management.

BACKGROUND: The association of bowel atresia with gastroschisis is well recognized, but the ideal management is less certain. METHODS: The records of 10 infants with gastroschisis and intestinal atresia treated between 1991 and 1997 in a single neonatal surgical unit were reviewed. RESULTS: Ten infants had midgut atresias: 5 small intestine, 1 jejunum and colon, and 4 colonic. Of the 6 with small bowel atresias, 4 had primary abdominal wall repair with electively delayed primary anastomosis at 21 to 46 days, 1 had decompressing tube jejunostomy at 11 days at the time of secondary abdominal closure, and 1 initial jejunostomy. Of the 4 with colonic atresia alone, 3 had perforation or infarction of the proximal colon at birth, 2 had a primary stoma, and 2, who did not have initial colostomy or anastomosis, required reoperation for increasing abdominal distension at 11 and 23 days. Complications were recorded in 6 patients: 2 had necrotizing enterocolitis after which 1 developed malabsorption, 1 had distal bowel obstruction after delayed primary anastomosis treated with a Bishop-Koop ileostomy at 50 days, 2 had stoma stenosis, and 1 had stoma prolapse. None died, and at 1 year 9 were within the normal range for body weight, and details of 1 are not known. CONCLUSIONS: At birth, diagnosis of atresia often is uncertain, and the bowel is difficult to repair or exteriorize. For proximal atresias, delayed primary repair is a safe satisfactory approach. At birth, distal atresias are more obvious, often complicated by perforation or infarction, and may benefit from early enterostomy.

Duodenal leiomyosarcoma presenting with iron deficiency anemia.

Failed iron therapy warrants investigation to rule out disorders of iron absorption or intestinal blood loss. The authors report the third case of duodenal leiomyosarcoma in childhood, which presented with iron deficiency anemia. Endoscopy failed to elicit the cause of this problem. Wide surgical resection, sparing the pancreas, was possible.

An autonomous module for supporting mice during spaceflight.

The Animal Module for Autonomous space Support (A-MASS) was developed to enable 30-day spaceflight for mice on the first Commercial Experiment Transporter mission. Because space hardware did not previously exist to support mice without astronaut intervention, the A-MASS presented considerable technical and animal care challenges. The technical challenges included maintaining a 42.5l payload volume and 20-g structural conformance while providing 30 days of autonomous mouse support. Sensors, video, a pressurized oxygen supply system and an internal data logging system were incorporated. The A-MASS met NIH guidelines for temperature, humidity, food and water access, oxygen supply, air quality and odor control. These technical and animal care challenges, along with power and mass constraints, were addressed using a novel design which ensures a fresh food and water supply, a clean view path into the cage for the camera system, and removal of the wastes from the air supply. The payload was successfully tested in an enclosed chamber and passed animal health, vibrational, mechanical, and electrical tests. The physiological, tactical and animal support information gathered will be applicable to the development of mouse support modules for the Shuttle Middeck and Space Station Freedom Express Rack environments.

Communication in and clinician satisfaction with multidisciplinary team meetings in neuro-oncology.

Multidisciplinary Team (MDT) meetings are critical in the management of complex cancer cases. There are limited data regarding the effectiveness of neuro-oncology MDT meetings and the impact of documenting and disseminating the recommended patient management. We established a weekly neuro-oncology MDT meeting and developed a standard electronic communication process. A survey was issued to participating clinicians to assess their level of satisfaction. The survey revealed that 100% felt the meeting and its documentation was very or extremely important, and 94% (n=15) felt the meeting was effective in documentation and communication of plans. There was a mixed response regarding which patients should be discussed: 44% (n=7) thought all patients should be discussed and 56% (n=9) thought only those patients with complex management issues should be discussed. We have developed an efficient method of documenting and disseminating patient information arising from our neuro-oncology MDT meeting. Clinician satisfaction was high.

Cullen's sign and massive ovarian enlargement secondary to primary hypothyroidism in a patient with a normal FSH receptor.

Ovarian hyperstimulation is a recognised complication of longstanding hypothyroidism. A 12 year old girl with atrophic thyroiditis who presented with abdominal pain and distension is reported. She was noted to have bruising in the vicinity of the umbilicus (Cullen's sign). She had pronounced ovarian enlargement on ultrasonography and it was hypothesised that this profound phenotype might reflect an abnormal FSH receptor. However sequencing of the FSH receptor was normal. The ovarian enlargement resolved with thyroxine replacement. Physicians and surgeons should consider longstanding hypothyroidism in patients presenting with Cullen's sign.

Repeat survey of current practice regarding corticosteroid prophylaxis for patients at increased risk of adverse reaction to intravascular contrast agents.

AIM: To compare the findings of a survey undertaken by us in 2002 regarding steroid premedication given in radiology departments to reduce the risk of adverse reactions in patients at increased risk of intravascular contrast media reactions with a similar survey published in 1994 by R. Seymour et al. The high risk patients considered in our survey were patients with history of asthma, drug allergies, hay fever and eczema. MATERIALS AND METHODS: A total of 225 questionnaires were sent to the consultant in charge for audit for radiology departments from the list given by the Royal College of Radiologists. 175 of the 225 questionnaires were returned (response rate 77.8%) and of these 172 were analysed with respect to the type, dose and duration of steroids. RESULTS: Compared to the survey in 1994, it was found that the number of departments who use steroid cover for all category of risk factors had increased compared to previous survey (73.8% in 2002 versus 55.3% in 1994 (p=0.001). There is now almost universal use of non-ionic contrast 98.8% versus 82.4% in 1994 (p=0.001). There is no agreed policy among radiology departments for the need or the dose or duration of steroid cover. CONCLUSION: Despite the more widespread use of non-ionic contrast media, the use of steroid premedication has increased which is contrary to what is expected as the incidence of adverse reaction to non ionic media is less than ionic contrast media.

Pigmented spots in the wool-bearing skin on white merino sheep induced by ultraviolet light.

Black-grey pigmented skin spots, some of which contained pigmented wool fibres, were observed in a flock of 8.5-year-old white Merino ewes. The spots were concentrated along the backline and increased in number following shearing, suggesting exposure to sunlight to be of importance in the development of these non-congenital pigmented skin spots in genetically white Merino sheep. To test the effect of ultraviolet light, white Merino sheep, ranging in age from 3 to 8 years, had a closely clipped midside area of wool-bearing skin irradiated on each of 28 consecutive days. Pigmented skin spots developed in 6 of the 16 white Merino sheep irradiated. Spots first appeared after 10 days of irradiation, the number subsequently increasing with time, and two skin spots were found to contain sparse numbers of black-grey pigmented wool fibres. Histological examination showed both the naturally occurring and irradiation-induced pigmented skin spots resulted from an increase in both number and activity of melanocytes localized along the epidermal-dermal border of the epidermis. With time, the melanocytes were observed to have entered, to varying depths, the outer-root sheath of follicles still producing white wool fibres. These ultraviolet-light-induced changes to epidermal melanocytes in white Merino sheep presumably occur due to alterations within the local tissue environment in which the melanocytes lie.

Albinism in a Suffolk sheep.

This report introduces the second form of true albinism to be documented in sheep, which appears mild enough not to cause serious undesirable side effects yet apparently effective enough to have the potential for general usage in the sheep industry. Based on the matings conducted to date, the albinism is inherited like an autosomal recessive. Histochemical tests reveal a defective melanin synthesis involving a block to the conversion of tyrosine to dopa but not the subsequent reactions that lead to melanin. The enzyme tyrosinase is a product of the C locus and catalyzes the conversion of tyrosine to dopa and the following reaction (dopa to dopaquinone). Therefore, it is proposed that the albinism arises from a gene in the C locus that encodes a defective tyrosinase. The gene is provisionally named albino marrabel, the gene symbol is cmar, and the locus allele symbol is Ccmar.

Dorsal relieving incision for the non-retractile foreskin.

The operation of dorsal relieving incision (DRI) for non-retractibility of the foreskin is described and the results of a retrospective review of 67 boys so treated are presented. We conclude that DRI is a simple, safe and effective procedure for the non-retractile foreskin not affected by fibrotic scarring.

Subclavian artery injury following percutaneous insertion of dialysis catheter.

Iatrogenic subclavian artery injury is a rare but recognized complication of central venous catheterization. The lesion is more severe and complex to treat when produced by large catheters such as dialysis catheters. A case is presented below with a discussion of vascular access for renal replacement therapy and the principles of treating penetrating arterial trauma at this location.