Sex pheromone of the winter moth, a geometrid with unusually low temperature precopulatory responses.

The sex pheromone for the winter moth, Operophtera brumata (L.), has been identified as the novel compound (Z,Z,Z)-1,3,6,9-nonadecatetraene. The male moths respond to the pheromone at low temperatures (4 degrees to 15 degrees C) and exhibit an upper response limit that coincides with the lower response limit for other reported moth sex pheromone systems. The pheromone attracted two other geometrid species, O. bruceata (Bruce spanworm) and O. occidentalis.

Identification and molecular characterisation of a CALM-AF10 fusion in acute megakaryoblastic leukaemia.

The t(10;11)(p13;q14-21) is a non-random translocation described in acute lymphoblastic and myeloid leukaemias. It results in the fusion of the gene CALM, which encodes a clathrin assembly protein, on 11q14 to the gene AF10, a putative transcription factor on 10p13. Here we describe for the first time, the occurrence of a CALM-AF10 fusion in a case of acute megakaryoblastic leukaemia. Fluorescence in situ hybridisation and reverse transcriptase polymerase chain reaction were used to confirm the presence of a CALM-AF10 fusion. A novel splice variant of CALM missing nt 1927-2091 was also detected. Though CALM is a cytoplasmic protein, the chimaeric fusion product is able to localise to both the nucleus and cytoplasm. Analysis of the fusion variants suggests, however, that the critical fusion product is likely to be cytoplasmic and contain the interactive leucine zipper of AF10.

Squamous oesophageal cancer can be downstaged using protracted venous infusion of 5-fluorouracil with epirubicin and cisplatin (ECF).

21 patients with squamous oesophageal carcinoma were treated with a new regimen designed in our unit and effective in treating gastric adenocarcinoma, consisting of continuous venous infusion of 5-fluorouracil for up to 24 weeks (200 mg/m2/day) with epirubicin (50 mg/m2) and cisplatin (60 mg/m2) every 3 weeks. 12 patients (57%) had an objective response. The median relapse free period was 7 months, median survival from start of chemotherapy 8.4 months, and median survival from diagnosis, 14 months. Symptomatic improvements were reported by 10/11 patients with pain (91%), 8/9 with anorexia (89%), 8/10 with reflux (80%) and 10/14 with dysphagia (71%). Grade 3 or 4 toxicity was reported by 11 patients: 5 had haematological toxicity, 3 vomiting, 2 infection and 1 diarrhoea. One patient developed peripheral neuropathy, 1 renal impairment and another peripheral vascular disease. Following chemotherapy, surgery was attempted in 5 patients. One remains well 3 years on, 2 had macroscopic clearance of tumour but died of postoperative complications. In 2, disease was irresectable. This regimen of moderate toxicity is effective at improving symptoms in the majority of patients. In some patients, tumours are briefly downstaged so that inoperable tumours may become operable.

Altering the fine specificity of an anti-Legionella single chain antibody by a single amino acid insertion.

Antibody engineering provides the potential to clone and manipulate antibody genes to produce fragments with altered specificity. We have produced an anti- Legionella single chain fragment with broader specificity towards Legionella serotypes than the parent monoclonal antibody. Using this relationship between the parent monoclonal and the recombinant antibody derived from it as a model, we attempted to identify those residues responsible for this change in fine specificity. Sequence analysis of this recombinant antibody revealed the deletion of a conserved residue, Asp101, in the CDR-H3 region. Using site-directed mutagenesis, we have created a mutant form of this single chain fragment with an aspartic acid insertion mutation at position 101 of the antibody heavy chain. This mutant scFv demonstrates improved specificity compared to the wild-type recombinant antibody, indicating an important role for Asp101.

Malignant lymphoma of the cervix uteri: histology and ultrastructure.

Two cases of primary lymphoma of the cervix uteri are described. Both responded to radiotherapy; both were composed at the ultrastructural level of mature macrophages and immature, apparently neoplastic lymphoreticular cells and are classified as reticulum cell lymphoma.

C-myc oncogene product P62c-myc in ovarian mucinous neoplasms: immunohistochemical study correlated with malignancy.

The monoclonal antibody Myc 1-6E10 was used to determine the cellular distribution of the c-myc oncogene product p62c-myc in 60 mucinous ovarian tumours. Three patterns of immunostaining were apparent: (i) nuclear staining alone; (ii) staining of the nucleus and basal cytoplasm; and (iii) staining of the entire cell. Of the 21 cases of mucinous cystadenoma, 11 showed nuclear staining alone, and a further case showed additional weak staining of the basal cytoplasm. Nuclear staining alone was not present in any of the 17 borderline mucinous tumours examined. Strong staining of the nucleus and basal cytoplasm was seen in 16 of these borderline cases, six of which also showed focal staining of the apical cytoplasm. All 22 cases of mucinous cystadenocarcinoma showed staining of the cell nucleus and entire cell cytoplasm. Focal staining of the apical cytoplasm in six of 17 borderline mucinous tumours produced a pattern of c-myc immunostaining similar to that of cystadenocarcinoma. Retrospective analysis of the clinical data showed that no significant differences between patients with borderline tumours of these two categories could be defined. Although immunostaining with Myc 1-6E10 can be used in the categorisation of mucinous ovarian tumours, it is concluded that standard histological criteria are more accurate indicators of tumour behaviour than is an assessment of c-myc expression.

Can histopathologists reliably diagnose molar pregnancy?

AIMS: To assess the degree of difficulty in diagnosing partial mole by analysing intraobserver and interobserver agreement among a group of pathologists for these diagnoses. METHODS: Fifty mixed cases of partial mole, complete mole, and non-molar pregnancy were submitted to seven histopathologists, two of whom are expert gynaecological pathologists; the other five were district general hospital consultants, one of whom works in Australia. These participants gave each slide a firm diagnosis of either partial mole, complete mole, or non-molar pregnancy. Some 12 months later, the slides were recorded and again submitted for a second diagnostic round to assess intraobserver as well as interobserver agreement. Standard histological criteria for each diagnostic category were circulated with the slides. RESULTS: kappa statistics showed that complete mole could be reliably distinguished from non-molar pregnancy, but neither non-molar pregnancy nor complete mole could be easily differentiated from partial mole. In only 35 out of 50 cases was there agreement between five or more of the seven participants. Agreement between the expert gynaecological pathologists was no better than for others in the group. Interestingly, the intraobserver agreement for each pathologist was good to excellent. CONCLUSIONS: These results imply that the reported histological criteria are either not being applied consistently or that they are lacking in practical use. An atypical growth pattern of trophoblast, rather than the polar accentuation seen in normal first trimester pregnancies, seems to be the important diagnostic histological feature for partial mole. Ploidy studies might also help with problem cases.

Chemistry, coeliac-toxicity and detection of gluten and related prolamins in foods.

Some recent advances in the understanding of the chemistry of gluten proteins and its relationship to the toxicity of different fractions in coeliac disease (gluten intolerance) is reviewed. Most recent studies on gluten toxicity have used in vitro analyses of cellular immune activation by gluten fractions and peptides. Our work indicates that gliadin is the most active of the different protein families found within the wheat grain and that a specific peptide sequence located in the amino terminus domain of alpha-gliadin and containing the sequence proline-serine-glutamine-glutamine was most active. Improvement in the dietary management of coeliac disease is possible by use of test kits for the detection of gluten in foods. Both laboratory kits and home test kits (suitable for use by individual coeliacs) are available and reliably detect gluten from wheat, rye and barley even after cooking or baking.

Immunoassay for wheat processing quality: utilization of a sandwich assay incorporating an immobilized single-chain fragment.

A single-chain fragment (scFv) was engineered from a monoclonal antibody to high molecular weight glutenin subunits (HMW-GS), wheat flour polypeptides that play a major role in determining the mixing- and extension strength-related properties of dough and its subsequent baking performance. The scFv was expressed in a thioredoxin mutant Escherichia coli strain that allows disulfide bond formation in the cytoplasm and incorporated into a diagnostic test for wheat quality. Although the scFv lacks the more highly conserved antibody constant regions usually involved with immobilization, it was able to be directly immobilized to a polystyrene microwell solid phase without chemical or covalent modification of the protein or solid phase and utilized as a capture antibody in a double-antibody (two-site) immunoassay. In the sandwich assay, increasing HMW-GS concentrations produced increasing assay color, and highly significant correlations were obtained between optical densities obtained in the ELISA using the scFv and the content of large glutenin polymers in flours as well as measures of dough strength as measured by resistance to dough extension in rheological testing. The assay using the scFv was able to be carried out at lower flour sample extract dilutions than that required for a similar assay utilizing a monoclonal capture antibody. This research shows that engineered antibody fragments can be utilized to provide superior assay performance in two-site ELISAs over monoclonal antibodies and is the first application of an engineered antibody to the analysis of food processing quality.

Adaptive transparent film dressings.

Transparent film dressings have many of the attributes of the ideal wound dressing. However, currently available film dressings are deficient in their ability to handle varying levels of wound exudate. The permeability of polymeric films to water vapor is discussed and techniques are described to produce films in which the moisture vapor permeability is a function of the moisture in the environment. Illustrations are provided showing the variation of permeability with relative humidity and water contact. The unique properties of coextruded films are illustrated and the responsiveness of such a film dressing to varying conditions at the wound are discussed.

Antioxidant prevention of Heinz body formation and oxidative injury in cats.

OBJECTIVE: To determine the effectiveness of 3 antioxidants in preventing Heinz body anemia in cats. DESIGN: Prospective study. ANIMALS: 44 specific-pathogen-free healthy cats. PROCEDURE: Cats were housed individually, divided randomly into 4 groups, and given the following orally every 12 hours: empty gelcaps (control cats), N-acetylcysteine (NAC, 100 mg/kg of body weight), vitamin E (d,l-alpha-tocopherol; 400 IU), or ascorbate (250 mg). After 2 weeks, Heinz bodies were induced by dietary onion powder (OP; 1% or 3% of dry matter) or propylene glycol (PG, 8% wt/vol in drinking water) for an additional 3 weeks. Intake of treated water or food was recorded daily. Body weight, PCV, Heinz body and reticulocyte percentages, reduced glutathione concentration, and total antioxidant status were measured twice weekly in all cats. RESULTS: Heinz body percentage and degree of anemia did not differ significantly among cats receiving antioxidants and control cats except in cats that ingested water containing PG, in which antioxidant supplementation was associated with a decrease in water intake. Of cats that were fed a diet that contained OP, cats that received NAC had significantly higher reduced glutathione concentrations, compared with other cats in the experiment. Total antioxidant status did not consistently correlate with antioxidant supplementation or type of oxidant administered (ie, OP or PG). CONCLUSIONS AND CLINICAL RELEVANCE: Although the effect of antioxidant supplementation on Heinz body anemia in cats was minimal, antioxidants may have subclinical biochemical effects such as GSH sparing that may be important against milder forms of oxidative stress.

Rapid antibody-based field test to distinguish between Helicoverpa armigera (Lepidoptera: Noctuidae) and Helicoverpa punctigera (Lepidoptera: Noctuidae).

Helicoverpa armigera (Hübner) and Helicoverpa punctigera (Wallengren) are the two most important insect pests of cotton production in Australia and require application of insecticides to control them. H. armigera has developed resistance to several insecticides but H. punctigera has not. Cost-effective management of insecticide resistance requires that growers be able to determine the proportion of H. armigera eggs or young larvae present on their crop before applying insecticides. This is impossible visually. We generated two monoclonal antibodies that reacted with the insect protein "lipophorin" and were capable of discriminating individuals of the two species at all life-stages. The antibodies were incorporated into a rapid test kit that was tested under field conditions over two growing seasons. Results obtained with the kit agreed closely with those obtained by rearing larvae through to second instar.

Precepting the clinical nurse specialist student.

The education role of the clinical nurse specialist (CNS) includes precepting the CNS graduate student. If the novice CNS is left to flounder, preceptorship can be a frustrating experience. This article describes a model for precepting CNS graduate students. Such a model provides a guideline and a sense of direction for the CNS. To clarify further the role of the CNS as a preceptor, the article offers practical learning experiences for the implementation of the model.

Introduction of solid foods to African American and Anglo American low-birth-weight and full-term infants.

A secondary data analysis of 7,174 infants explores the use of cereal, fruits and vegetables, and meats with African-American and Anglo-American very-low-birth-weight (VLBW), low-birth-weight (LBW) and term infants over the first five months after discharge. The first solid foods offered were cereal for African-American infants and fruits and vegetables for Anglo infants. There are statistically significant differences in the number of feedings offered by the two ethnic groups. During the last two-to-three months, African-American term and LBW infants received all solid foods more frequently than Anglo infants. Neither ethnic group followed current feeding practice recommendations on when to introduce solid food.

Allelic association and recombination hotspots in the mucin gene (MUC) complex on chromosome 11p15.5.

A family of four highly polymorphic genes encoding secreted gel forming mucins is located in the middle of a recombination rich region of the short arm of chromosome 11 (11p15.5; tel MUC6-MUC2-MUC5AC-MUC5B cen; approx. 400 kb). These genes are of interest as risk factors for inflammatory diseases of the epithelia, and we have for example reported association of a VNTR polymorphism of the major mucin domain of MUC2 with asthma, despite the fact that MUC2 is not a major respiratory mucin. To understand the significance of this and other mucin gene associations it is important to describe the patterns of linkage disequilibrium (LD) across this chromosomal region, which is still incomplete on HapMap and the UCSC Golden Path sequence. Our previous studies on the 40 core CEPH families provided direct evidence for several recombination events within the immediate region of the gene complex. This study examines these recombination events in more detail, and also the patterns of LD across the gene complex. We refine the location of the breakpoints, and the combined data suggest two probable recombination hotspots. Three breakpoints are located between MUC6 and MUC2: there is no association between MUC6 and MUC2, and the data collected here, combined with that publicly available, maps a hotspot to a region of 4 kb. The other recombinants map between MUC2 and intron 8 of MUC5B. Relatively strong LD is detected between MUC2 and MUC5AC, and although 10/70 of the chromosomes tested shared a common haplotype, which extends from MUC2 to MUC5B, statistically significant association was not detected between MUC2 and the markers tested in MUC5B. We discuss the possibility that the previously reported association between MUC2 and asthma is most likely attributable to association with functional variation in MUC5AC, which encodes one of the two major mucins expressed in both healthy and diseased airways.

Preliminary findings: a maximum oral feeding time for premature infants, the relationship to physiological indicators.

Oxygen saturation (SaO2) values, pulse rate and respiratory rate (RR) were monitored using a pulse oximeter during feeding sessions for 21 preterm infants (mean postconceptual age 35.61 weeks) at 3-minute intervals. The purpose of the study was to determine if there is a maximum stressful nippling time span within the context of the prescribed amounts of formula taken and to determine and verify the relationship between: (a) nipple feeding and clinical stress reactions to these variables, (b) the amount of formula taken in the first 3 minutes and (SaO2) values, PR, and RR; and (c) the birthweight of the infant and the length of time from birth to the initial nipple feed. Although the repeated analysis of variance failed to show a significant change (p > .05) from baseline in SaO2, PR, and RR, correlational analysis showed a significant relationship between the amount of formula taken in the first 3 minutes of feeding and the respiratory rate (r = .69, p = .001). Since no significant declines were detected across time in the physiological measures, a maximum stressful nippling time was not established from these data. As expected, a significant relationship between birthweight and the length of time from birth to the initial nipple feed was revealed (p < .05).

Enzyme immunoassay for determination of gluten in foods: collaborative study.

A collaborative study was performed in 15 laboratories to validate a monoclonal antibody-based enzyme immunoassay (EIA) for determination of gluten in foods. The study included 13 samples: maize starch, "gluten-free" baking mixes, wheat flours, cookies, cooked meats, and a soup. Gluten was present in these samples at either zero or 0.02 to 10% by weight, i.e., over almost 3 orders of magnitude. The mean assay values for the foods varied from 88 to 105% of the actual amounts. The assay was quantitative for cereal products and the soup with repeatability (RDS-r, relative standard deviation) and reproducibility (RSD-R) of 16-22% and 24-33%, respectively. The assay was semiquantitative for the processed meat products (RSD-r 14 and 26% and RSD-R 46 and 56%), probably because gluten was unevenly distributed in the small (1 g) samples that were analyzed. The ELISA method produced no false positive results, and false negatives obtained with tannin-containing foods could be avoided by use of a modified sample extractant. None of the collaborators reported problems in following the protocol. The method has been adopted official first action by AOAC for determination of wheat gluten in foods.