Setting Scottish diagnostic reference levels for mammography incorporating both craniocaudal and oblique projections between 30 and 80 mm.

Optimisation must be carried out on all medical radiological units to ensure doses are as low as reasonably practicable, consistent with the intended purpose. To achieve this, population doses must be estimated and diagnostic reference levels (DRLs) set. For mammography examinations, mean glandular doses (MGDs) are calculated for this purpose. The average MGD per unit is compared to the national mammography DRL, which is applicable to compressed breast thicknesses (CBTs) of 50-60 mm for oblique (OB) views only and set using data from screening units. It is the purpose of this work to assess planar MGDs across Scotland and set DRLs based on data collected from all screening and symptomatic units across Scotland, considering craniocaudal (CC) and OB views and a wider range of CBTs. Data from the most recent dose audit (spanning 2015-2017) for 67 mammography x-ray units were collated and analysed (26 195 images). No large differences between MGD of CC and OB views were found when considering specific CBT ranges (median difference 2.6%). There was, however, a significant difference between screening and symptomatic data (19%). As expected, MGD increased with CBT and there were significant differences in MGD between manufacturers. From the data analysed, Scottish DRLs were set based on 95th percentile values for digital mammography units for three CBT ranges (30-49, 50-60 and 61-80 mm): 1.3, 1.8 and 2.6 mGy respectively. These values consider OB and CC views collectively. Fifth percentile values are quoted to highlight units at greater risk of insufficient image quality. These MGD values, together with image quality assessments, will facilitate optimisation across Scotland. Results show that use of different CBT ranges and inclusion of CC views increases the number of images included in dose audit data analysis from approximately 12%-92%, which is substantially more representative of the population.

Resting metabolic rate prediction equations and the validity to assess energy deficiency in the athlete population.

NEW FINDINGS: What is the topic of this review? We review the issues with using predicted resting metabolic rate equations in athletic populations. What advances does it highlight? The use of dated predicted resting metabolic rate equations is not appropriate for athletic populations until more studies have been conducted among these unique populations. ABSTRACT: Resting metabolic rate (RMR) is the amount of energy the body uses at rest. A suppressed RMR has been correlated with low energy availability and therefore used as an indicator of an individual's energy state. Furthermore, confounding identification of low energy availability within an athletic population are the physiological measures required, which can be time consuming and require professional expertise. To negate the demands of laboratory protocols in measuring RMR, predicted RMR (p RMR) equations were developed. Caution should be exercised when applying the p RMR equations for determining low energy availability in athletes owing to the population used to develop the equations and the higher metabolic cost of fat-free mass, thus elevated RMR, associated with athletes. Moreover, a low ratio of measured RMR to p RMR is often used as an alternative marker for energy deficiency. Predictive equations should implement fat-free mass within the algorithm when estimating RMR in athletic populations. The purpose of this paper is to describe p RMR equation development and the issues associated with use of p RMR equations for athletic populations. As professional sport increases, validation of p RMR equations in the modern athlete population is needed to monitor energy availability for athletic health and performance.

Fibronectin-binding protein A of Staphylococcus aureus has multiple, substituting, binding regions that mediate adherence to fibronectin and invasion of endothelial cells.

Invasive Staphylococcus aureus infection frequently involves bacterial seeding from the bloodstream to other body tissues, a process necessarily involving interactions between circulating bacteria and vascular endothelial cells. Staphylococcus aureus fibronectin-binding protein is central to the invasion of endothelium, fibronectin forming a bridge between bacterial fibronectin-binding proteins and host cell receptors. To dissect further the mechanisms of invasion of endothelial cells by S. aureus, a series of truncated FnBPA proteins that lacked one or more of the A, B, C or D regions were expressed on the surface of S. aureus and tested in fibronectin adhesion, endothelial cell adhesion and invasion assays. We found that this protein has multiple, substituting, fibronectin-binding regions, each capable of conferring both adherence to fibronectin and endothelial cells, and endothelial cell invasion. By expressing S. aureus FnBPA on the surface of the non-invasive Gram-positive organism Lactococcus lactis, we have found that no other bacterial factor is required for invasion. Furthermore, we have demonstrated that, as with other cell types, invasion of endothelial cells is mediated by integrin alpha5beta1. These findings may be of relevance to the development of preventive measures against systemic infection, and bacterial spread in the bacteraemic patient.

Country report: Australia.

The last 10 years has been an interesting time for Australian medical education despite reduced funding. WORKFORCE: There are five main workforce trends: a rural/urban maldistribution, a need for more specialists, public hospital staffing difficulties, increasing female practitioners and under-representation of indigenous practitioners. ISSUES FACING THE DEANS: Lack of resources is a problem facing Deans, with pressure for clinical service in teaching hospitals. Entrepreneurial activities have been undertaken including the enrollment of overseas students. Medical schools have also responded to important government initiatives. DEVELOPMENTS IN MEDICAL EDUCATION: Australia's 11 medical schools have undergone significant reform in the last decade. There is a mix of four (graduate), five and six year courses. AUSTRALIA'S NEW MEDICAL SCHOOL: James Cook University opened the first medical school in northern Australia in 2000. The School admits students from rural, northern Australian and indigenous backgrounds. It has a strong regional mission. RURAL AND COMMUNITY-BASED EDUCATION: Government funding to address the maldistribution of the workforce has led to the establishment of rural clubs, Departments of Rural Health and community-based programs. THE FIRST TWO POSTGRADUATE YEARS: There have been recent moves to improve education in the two years following graduation. This includes the initiation of national projects in curriculum and assessment. POSTGRADUATE AND CONTINUING MEDICAL EDUCATION: Postgraduate programs in Australia are being reformed to build on the changes in undergraduate education. CME is also under review. CONCLUSION: Australian medical educators should build on the recent reforms and take on some of the new directions in medical education.

E-cadherin expression is a sensitive and specific method for detection of carcinoma cells in fluid specimens.

The distinction between reactive mesothelial cells and carcinoma in pleural, peritoneal, and pericardial fluids is often difficult. We and others previously showed that E-cadherin, an epithelial-specific adhesion protein, can be useful for this distinction. In this study we tested the sensitivity and specificity of E-cadherin compared to, and in combination with, conventional cytology for assessment of carcinoma in fluids. Cytyc ThinPreptrade mark slides (Marlborough, MA) from 102 sequential fluids were evaluated for E-cadherin expression by routine immunologic techniques. No evidence of E-cadherin staining was seen in 71 cases, while 31 showed unequivocally positive staining. Sensitivity and specificity were independently determined for E-cadherin alone (72% and 97%, respectively), cytomorphology alone (62% and 100%, respectively), and both together (92% and 100%, respectively). We conclude that assessment of E-cadherin expression has sensitivity and specificity comparable to, or better than, conventional cytomorphology. If both cytomorphology and E-cadherin are used together, a definitive and correct diagnosis could have been made on nearly every case in this study.

Comparison of acoustic and perceptual measures of voice in male-to-female transsexuals perceived as female versus those perceived as male.

The present study explored significant differences between male-to-female transgendered speakers perceived as male and those perceived as female in terms of speaking fundamental frequency (SFF) and its variability, vowel formants for /a/ and /i/, and intonation measures. Fifteen individuals who identified themselves as male-to-female transsexuals served as speaker subjects, in addition to 6 biological female control subjects and 3 biological male control subjects. Each subject was recorded reading the Rainbow Passage and producing the isolated vowels /a/ and /i/. Twenty undergraduate psychology students served as listeners. Results indicated that subjects perceived as female had a higher mean SFF and higher upper limit of SFF than subjects perceived as male. A significant correlation between upper limit of SFF and ratings of femininity was achieved.

Expression of proteoglycan core proteins in human bone marrow stroma.

Heparan sulphate proteoglycans (HSPGs) present on the surface of bone marrow stromal cells and in the extracellular matrix (ECM) have important roles in the control of adhesion and growth of haemopoietic stem and progenitor cells. The two main groups of proteoglycans which contain heparan sulphate chains are members of the syndecan and glypican families. In this study we have identified the main surface membrane and matrix-associated HSPGs present in normal human bone marrow stroma formed in long-term culture. Proteoglycans were extracted from the adherent stromal layers and treated with heparitinase and chondroitinase ABC. The core proteins were detected by Western blotting using antibodies directed against syndecans-1-4, glypican-1 and the ECM HSPG, perlecan. Stromal cell expression at the RNA level was detected by Northern blotting and by reverse transcription PCR. Glypican-1, syndecan-3 and syndecan-4 were the major cell-membrane HSPG species and perlecan was the major ECM proteoglycan. There was no evidence for expression of syndecan-1 protein. Syndecan-3 was expressed mainly as a variant or processed 50-55 kDa core protein and in lower amounts as the characteristic 125 kDa core protein. These results suggest that syndecan-3, syndecan-4 and glypican-1 present on the surface of marrow stromal cells, together with perlecan in the ECM, may be responsible for creating the correct stromal 'niche' for the maintenance and development of haemopoietic stem and progenitor cells. The detection of a variant form of syndecan-3 as a major stromal HSPG suggests a specific role for this syndecan in haemopoiesis.

alpha4beta1 integrin-mediated adhesion of CD34+ cells from patients with chronic myeloid leukaemia: influence of IL-3.

Interactions between integrins on haemopoietic progenitor cells and their stromal ligands have an important role in the control of haemopoiesis. Growth factors can modulate these interactions (so-called 'inside-out' signalling) resulting in changes in ligand binding activity. We have studied alpha4beta1 integrin-mediated adhesion to the H120 fragment of fibronectin (which contains the strongest alpha4beta1 binding site) in CD34+ cells from patients with chronic myeloid leukaemia (CML) and have determined the effect of IL-3 on the level of adhesion. Compared to normal CD34+ cells isolated from cord blood and peripheral blood progenitor harvests (mean of 61.4 +/- 14.9% of cells attached) the CML CD34+ cells showed reduced levels of adhesion (mean of 41.9 +/- 14.7%, P < 0.05). The effect of 10 ng/ml of IL-3 resulting in reduced adhesion of normal CD34+ cells at 30 min was absent in 6/7 patients with CML. Abnormalities of adhesion to fibronectin may thus be related to IL-3 pathways affected by BCR-ABL. These findings will have implications for understanding the dysregulation of growth and adhesion in CML.

Coordinated modulation of the fibroblast growth factor dual receptor mechanism during transformation from human colon adenoma to carcinoma.

Basic fibroblast growth factor (bFGF) is dependent on heparan sulphate for its ability to activate the cell surface signal transducing receptor. We have investigated the FGF dual receptor mechanism in a novel model of the transformation from human colon adenoma to carcinoma in vitro. Reverse transcription-polymerase chain reaction showed that mRNA for FGF receptors 1 and 2 were expressed in both the adenoma and carcinoma cells whereas immunocytochemistry showed that the expression of the FGF R1 was reduced significantly in the carcinoma cells. We have reported previously that the composition and sequence of human colon adenoma and carcinoma heparan sulphate (HS) differ in a defined and specific manner. The functional significance of these changes was assessed by affinity co-electrophoresis, which showed that the affinity of adenoma HS for bFGF was 10-fold greater than that of the carcinoma HS (Kd 220 nM vs. 2493 nM, respectively). In addition, Northern studies of the expression of syndecan 1 and 4 mRNA showed that proteoglycan core protein expression was reduced significantly in the carcinoma cells. These findings were associated with a reduced biological response to bFGF in the carcinoma cells that could be partially reversed by the addition of exogenous heparin, suggesting that both the proteoglycan and signal transducing receptor control the cells' response to bFGF.

Assessment of mucociliary transport in patients with chronic mucoid rhinitis.

Chronic rhinitis is the manifestation of a heterogeneous group of disease entities and often proves difficult to manage successfully. We present the investigations of the mucociliary system in 40 patients with mucoid rhinorrhoea as their principal symptom of whom 20 had pan respiratory disease. The saccharin clearance time (SCT) was measured and classified as normal if it was below 20 min. Objective measurement of clearance was made using 99mTechnetium-labelled human serum albumin (99mTc-HSA). We have standardized our method using a micrometer syringe driver to produce a droplet of consistent size (droplet size, 0.01 ml, SD 0.0002 ml) that reduces the dose of radiation. The movement of the droplet was measured over 20 min (RLT). The mean, maximum rate and percentage moved were calculated. Patients were divided into those who had chest disease (20) and those without and a chi 2-test was performed for the mean RLT time between the two groups. There was a strong correlation between mean and maximum rates (r = 0.91). One patient has a normal SCT and normal RLT. Patients with chest disease had a significantly lower mean RLT (P > 0.01). Assuming that RLT is the standard investigation, six patients were normal but had an abnormal SCT, this is a false positive error of 15%. The false negative error was 4/40 (10%). The association between sinus and chest disease with abnormal mucociliary clearance is stressed.

Transcription analysis of the prolate-headed lactococcal bacteriophage c2.

A detailed transcription map of the prolate-headed lactococcal phage c2 has been constructed. Transcription of about one-third of the genome, encoding 22 open reading frames, began within the first 2 min of infection and produced at least 12 overlapping transcripts that persisted until lysis occurred at 30 min after initiation of infection. The remaining two-thirds of the genome, encoding 17 open reading frames, was divergently transcribed, beginning between 4 and 6 min after initiation of infection, and resulted in at least 18 overlapping transcripts that persisted until lysis. Five very strong, simultaneously active, and probably unregulated early promoters and a single positively regulated late promoter were identified. The late promoter had an extended -10 sequence, had a significant basal level of activity in the uninduced state, and was induced to high activity by a phage gene product. The complex overlapping pattern of transcripts resulted from the action of the multiple early promoters, inefficient termination of transcription, and (possibly) processing of a late precursor transcript(s). Phage proteins were not required for these processes, and the host RNA polymerase was probably used for both early and late transcription.

Mucosal delivery of murine interleukin-2 (IL-2) and IL-6 by recombinant strains of Lactococcus lactis coexpressing antigen and cytokine.

Lactococcus lactis is a nonpathogenic and noncolonizing bacterium which is being developed as a vaccine delivery vehicle for immunization by mucosal routes. To determine whether lactococci can also deliver cytokines to the immune system, we have constructed novel constitutive expression strains of L. lactis which accumulate a test antigen, tetanus toxin fragment C (TTFC), within the cytoplasmic compartment and also secrete either murine interleukin-2 (IL-2) or IL-6. When mice were immunized intranasally with various different expression strains of L. lactis, the anti-TTFC antibody titers increased more rapidly and were substantially higher in mice immunized with the bacterial strains which secreted IL-2 or IL-6 in addition to their production of TTFC. This adjuvant effect was lost when the recombinant strains of L. lactis were killed by pretreatment with mitomycin C and could therefore be attributed to the secretion of IL-2 or IL-6 by the recombinant lactococci. These results provide the first example of the use of a cytokine-secreting, noninvasive experimental bacterial vaccine vector to enhance immune responses to a coexpressed heterologous antigen and point the way to experiments which will test the possible therapeutic efficacy of this mode of cytokine delivery.

The effect of alpha4 beta1-integrin binding sequences of fibronectin on growth of cells from human hematopoietic progenitors.

Highly regulated interactions between adhesion receptors on progenitor cells and their extracellular matrix ligands are essential for the control of hematopoiesis in bone marrow stroma. We have examined the relationship between alpha4beta1-integrin-mediated adhesion and growth of CD34(+) cells by assessing their adhesive and migratory patterns of proliferation in a mixture of hematopoietic growth factors in the presence of different recombinant fragments of the HepII/IIICS region of fibronectin. CD34(+) cells were isolated from cord blood and placed in culture wells containing serum-free medium and growth factors. Wells were precoated with either the H120 fragment of fibronectin, which contains three alpha4beta1-integrin binding sites, or the H0 fragment, which lacks the two highest affinity alpha4beta1 binding sequences. Proliferation of single cells of CD34(+)38(+)DR+ and CD34(+)38(-)DR+ phenotypes occurred in contact with the H120 substrate and was associated with migration. Larger numbers of cells were used to quantitate proliferative responses. Cells growing in wells coated with H120 formed attachments to the base of the wells throughout the culture period. Higher total cell counts were consistently found in wells coated with H120 compared with H0 and bovine serum albumin controls. The difference was first apparent at day 8 of culture and reached a maximum at days 11 through 13, when expansion with H120 was a mean of 1.8-fold higher than that seen with H0 (P

Maintenance of remission of ulcerative colitis: a comparison between balsalazide 3 g daily and mesalazine 1.2 g daily over 12 months. ABACUS Investigator group.

BACKGROUND: Despite widespread use of aminosalicylates as maintenance treatment for ulcerative colitis (UC), patients still report troublesome symptoms, often nocturnally. AIM: To compare the efficacy and safety of balsalazide (Colazide) with mesalazine (Asacol) in maintaining UC remission. METHODS: A randomized, double-blind comparison of balsalazide 3 g daily (1.04 g 5-ASA) and mesalazine 1.2 g daily for 12 months, in 99 (95 evaluable) patients in UC remission. RESULTS: Balsalazide patients experienced more asymptomatic nights (90% vs. 77%, P=0.0011) and days (58% vs. 50%, N.S.) during the first 3 months. Balsalazide patients experienced more symptom-free nights per week (6.4+/-1.7 vs. 4.7+/-2.8; P=0.0006) and fewer nights per week with blood on their stools or on the toilet paper, mucus with their stools or with sleep disturbance resulting from symptoms or lavatory visits (each P < 0.05). Fewer balsalazide patients relapsed within 3 months (10% vs. 28%; P=0.0354). Remission at 12 months was 58%, in both groups. Similar proportions of patients reported adverse events (61% balsalazide vs. 65% mesalazine). There were five serious adverse events (two balsalazide, three mesalazine) and four withdrawals due to unacceptable adverse events (three balsalazide, one mesalazine), of which one in each group was also a serious adverse event. CONCLUSIONS: Balsalazide 3 g/day and mesalazine 1.2 g/ day effectively maintain UC remission and are equally well tolerated over 12 months. At this dose balsalazide prevents more relapses during the first 3 months of treatment and controls nocturnal symptoms more effectively.

Influence of interleukin-3 and other growth factors on alpha4beta1 integrin-mediated adhesion and migration of human hematopoietic progenitor cells.

The mechanisms by which hematopoietic progenitor cells are normally anchored in stromal niches and yet can be mobilized by specific growth factors are poorly understood. It is likely, however, that integrins and their extracellular matrix (ECM) ligands play a key role in this process, and recent evidence suggests that integrin function is modulated by signals originating from activated growth factor receptors. We have now examined this further by studying the role of growth factors on alpha4beta1 integrin-mediated adhesion of human CD34+ hematopoietic progenitor cells to specific recombinant fibronectin fragments coated onto tissue culture dishes. Cells were prepared from cord blood and peripheral blood harvests. During a 30-minute adhesion assay a mean of 74% of CD34 cells attached to the so-called H120 fragment of fibronectin, which contains the strongest alpha4beta1 integrin-binding sequence. The level of cell adhesion was significantly reduced by low concentrations of interleukin-3 (IL-3) (2.5 to 10 ng/mL), whereas stem cell factor (SCF) and granulocyte colony-stimulating factor (G-CSF) at these concentrations did not affect adherence of the cells. Migratory behavior of CD34 cells was examined using fibronectin fragments adsorbed onto a Transwell filter. The H120 fragment supported much higher levels of cell migration than the H0 fragment of fibronectin, which contains a weak alpha4beta1 integrin binding sequence. Over a 16-hour assay, migration of peripheral blood progenitor cells was increased slightly by SCF and by G-CSF. However, a marked stimulation was observed with IL-3, which significantly increased migration. Similar effects were noted with cord blood cells, although a small proportion of cells were able to migrate in the absence of growth factors. These results indicate that there is a highly selective and functional link between the alpha4beta1 integrin and IL-3/IL-3-receptor that could affect the position of stem and progenitor cells in the marrow stroma and influence their growth and development.

Oral vaccination of mice against tetanus with recombinant Lactococcus lactis.

To determine whether a protective immune response could be elicited by oral delivery of a recombinant bacterial vaccine, tetanus toxin fragment C (TTFC) was expressed constitutively in Lactococcus lactis and administered orally to C57 BL/6 mice. The antibody titers elicited were lower than those following intranasal immunization (a route already known to result in high-level systemic anti-TTFC immune responses) but the protective efficacy was the same order of magnitude. The serum antibody isotypes elicited were predominantly IgG1 and IgG2a. TTFC-specific fecal IgA responses could be detected following oral or intranasal immunization. Chemically killed lactococci administered via the intranasal route were also able to elicit serum antibody responses of similar levels and kinetics to those induced by live bacteria.

The cell adhesion molecule, E-cadherin, distinguishes mesothelial cells from carcinoma cells in fluids.

BACKGROUND: The distinction between benign reactive mesothelial cells and well differentiated carcinoma can be difficult in pleural, peritoneal, and especially pericardial fluids. E-cadherin is an adhesion protein that is specifically expressed in cells of epithelial lineage. In this study, anti-E-cadherin antibodies were used to identify and distinguish carcinoma cells from reactive mesothelial cells. METHODS: Pleural, peritoneal, and pericardial fluids were prepared using the Cytyc Thin Prep processor. The specimens were comprised of a mix of 45 cases that were diagnosed as carcinoma, suspicious, or reactive by Papanicolaou staining of routine material seen by the authors' service. Routine immunologic techniques were used with a commercially available E-cadherin antibody. RESULTS: In most cases of carcinoma, tumor cells showed a strong positive membranous reaction product (32 of 37). This included four cases that were not cytomorphologically diagnosed as malignant, but subsequently proved to be malignant. E-cadherin staining was not observed in five tumors, two of which were not expected to express this protein. One benign case showed cells staining for E-cadherin, although the cells were not malignant by morphologic criteria. Because this case was a surgical pelvic washing, these cells more likely were epithelial contaminants than true false-positives. CONCLUSIONS: The epithelial specific cell-cell adhesion marker E-cadherin reliably distinguishes reactive mesothelial cells from carcinoma and is a useful adjunctive test to distinguish benign reactive mesothelial cells from well differentiated carcinoma cells in fluid specimens.