Frequent loss of RAF kinase inhibitor protein expression in acute myeloid leukemia.

RAF kinase inhibitor protein (RKIP) is a negative regulator of the RAS-mitogen-activated protein kinase/extracellular signal-regulated kinase signaling cascade. We investigated its role in acute myeloid leukemia (AML), an aggressive malignancy arising from hematopoietic stem and progenitor cells (HSPCs). Western blot analysis revealed loss of RKIP expression in 19/103 (18%) primary AML samples and 4/17 (24%) AML cell lines but not in 10 CD34+ HSPC specimens. In in-vitro experiments with myeloid cell lines, RKIP overexpression inhibited cellular proliferation and colony formation in soft agar. Analysis of two cohorts with 103 and 285 AML patients, respectively, established a correlation of decreased RKIP expression with monocytic phenotypes. RKIP loss was associated with RAS mutations and in transformation assays, RKIP decreased the oncogenic potential of mutant RAS. Loss of RKIP further related to a significantly longer relapse-free survival and overall survival in uni- and multivariate analyses. Our data show that RKIP is frequently lost in AML and correlates with monocytic phenotypes and mutations in RAS. RKIP inhibits proliferation and transformation of myeloid cells and decreases transformation induced by mutant RAS. Finally, loss of RKIP seems to be a favorable prognostic parameter in patients with AML.

Additional sex combs-like 1 belongs to the enhancer of trithorax and polycomb group and genetically interacts with Cbx2 in mice.

The Additional sex combs (Asx) gene of Drosophila behaves genetically as an enhancer of trithorax and polycomb (ETP) in displaying bidirectional homeotic phenotypes, suggesting that is required for maintenance of both activation and silencing of Hox genes. There are three murine homologs of Asx called Additional sex combs-like1, 2, and 3. Asxl1 is required for normal adult hematopoiesis; however, its embryonic function is unknown. We used a targeted mouse mutant line Asxl1(tm1Bc) to determine if Asxl1 is required to silence and activate Hox genes in mice during axial patterning. The mutant embryos exhibit simultaneous anterior and posterior transformations of the axial skeleton, consistent with a role for Asxl1 in activation and silencing of Hox genes. Transformations of the axial skeleton are enhanced in compound mutant embryos for the polycomb group gene M33/Cbx2. Hoxa4, Hoxa7, and Hoxc8 are derepressed in Asxl1(tm1Bc) mutants in the antero-posterior axis, but Hoxc8 expression is reduced in the brain of mutants, consistent with Asxl1 being required both for activation and repression of Hox genes. We discuss the genetic and molecular definition of ETPs, and suggest that the function of Asxl1 depends on its cellular context.

High expression of the sister-chromatid separation regulator and proto-oncogene hSecurin occurs in a subset of myeloid leukaemias but is not implicated in the pathogenesis of aneuploidy.

Aneuploidy is considered to play an important role in the pathogenesis of malignancies. We were interested whether abnormalities of the sister-chromatid separation regulator and proto-oncogene hSecurin occurred in myeloid leukaemias, and whether such abnormalities correlated with aneuploidy. The expression of hSecurin was assessed by real-time quantitative PCR in samples from patients with acute myeloid leukaemia (AML, n=70), chronic myeloid leukaemia (CML) in chronic phase (CP, n=20) or blast phase (BP, n=12), and granulocytes as well as mononuclear cells (MNCs) from healthy donors (n=21). Median hSecurin expression in AML with normal karyotypes was not significantly different from AML showing aneuploidy, CML BP or cells from healthy donors. However, hSecurin expression in CML CP was significantly increased compared to AML with normal karyotypes (1.82-fold; P<0.001), CML BP (3.18-fold; P<0.001), MNCs (3.17-fold; P<0.001) and granulocytes (2.69 fold; P<0.001) from healthy donors. Mutations in the coding region of hSecurin were not detected. These results do not support a major role of hSecurin in the development of aneuploidy in myeloid leukaemias. However, high expression of hSecurin may be of pathogenetic relevance in a subset of patients with regard to its potential to stimulate angiogenesis and to interact with the DNA-damage response pathway.

An application of the United Kingdom Working Party diagnostic criteria for atopic dermatitis in Scottish infants.

The United Kingdom Working Party diagnostic criteria for atopic dermatitis have been characterized in infants and children; however, the need for visual confirmation of flexural dermatitis by a trained investigator limits their use in large epidemiologic studies. We have administered the complete United Kingdom Working Party criteria in a postal questionnaire format to the mothers of year old infants and determined the concordance between mothers' and trained investigator's reports of visual flexural dermatitis. Based on mothers' responses to the questionnaire, 59 infants with atopic dermatitis and 59 controls were identified. In subsequent home interviews conducted by a trained investigator, the United Kingdom criteria questions were repeated and sites of current visible dermatitis were identified by mothers and the investigator as per United Kingdom Working Party protocol. Agreement between the mothers' postal and home interview responses was high: kappa= 0.75-0.94 for individual criteria; kappa= 0.93 for diagnosed atopic dermatitis. Agreement between the mothers' and investigator's observations of visible flexural dermatitis was high for all sites: kappa= 0.88-1.0. The results demonstrate that mothers are able to apply the United Kingdom criteria and accurately report visible flexural dermatitis in their year old infants. The postal application of the United Kingdom Working Party's diagnostic criteria for atopic dermatitis in year old infants appears to be a practical, reliable, epidemiologic tool in the investigation of atopic dermatitis with results comparable with formal application of the criteria by a trained investigator.

Intravenous administration of stabilized antisense lipid particles (SALP) leads to activation and expansion of liver natural killer cells.

Stabilized antisense lipid particles (SALP) have been developed for the systemic delivery of oligonucleotides. The impact of intravenous SALP administration was measured with respect to activation of natural killer (NK) and NK1.1+ T (NKT) cells in the livers of immunocompetent mice. Treatment with a SALP containing a highly mitogenic oligonucleotide (INX-6295) generated an increase in NK cytolytic activity and cell number within the liver but did not appear to affect the number of hepatic NKT cells or their cytolytic activity. The same results were observed after intravenous administration of the mitogenic oligonucleotide alone. Interestingly, treatment with a SALP containing a weakly mitogenic oligonucleotide (INX-6300) also activated the liver NK cells, whereas the oligonucleotide alone was unable to elicit these effects. The NK stimulatory activity of a SALP containing INX-6300 required both lipid and oligonucleotide components. These results demonstrate that in addition to modifying the pharmacokinetics and biodistribution of intravenously administered oligonucleotides, SALP possess immunostimulatory activity independent of oligonucleotide mitogenicity, which can serve as an adjuvant to antisense therapies for cancer.

Childhood exposure to infection and risk of adult onset wheeze and atopy.

BACKGROUND: The prevalence of asthma and allergic diseases in children and young adults is inversely associated with family size. It has been suggested that more frequent exposure to infections in a large family group, particularly those spread by the faecal-oral route, may protect against atopic diseases, although not all published data support this hypothesis. Whether similar considerations apply to adult onset wheeze is unknown. The relationship between adult onset wheezing and atopy measured in adulthood and childhood exposure to a range of infections was investigated. METHODS: A nested case control study of participants in a 30 year follow up survey was conducted. Questionnaire data on childhood infections had been obtained in a 1964 survey. In 1995 a further questionnaire on respiratory symptoms and other risk factors for wheezing illness was administered, total IgE, skin and RAST tests were performed, and serum was stored. In 1999 serological tests for hepatitis A, Helicobacter pylori, and Toxoplasma gondii were performed on the stored samples. Information from the 1964 questionnaires was available for 97 cases and 208 controls and serological tests were obtained for 85 cases and 190 controls. The potential risk factors were examined for all cases, those who reported doctor diagnosed asthma, those who described persistent cough and phlegm with wheeze, and subjects stratified by atopic status. RESULTS: The sibship structure was similar in cases and controls. In univariate analysis of all cases, childhood infections reported by parents as acquired either before or after the age of three years did not influence case:control or atopic status. Seropositivity was also similar for all cases and controls, but cases in the subgroup with chronic cough and phlegm were more likely to be seropositive for hepatitis A and H pylori. Seropositivity was unrelated to atopic status. In multivariate analyses both the effect of having two or more younger siblings (OR 0.1, 95% CI 0.03 to 0.8) and of acquiring measles up to the age of three (OR 0.2, CI 0.03 to 0.8) were significantly related to a lower risk of doctor diagnosed asthma. CONCLUSIONS: In these well characterised subjects, exposure to infections as measured by parental reports obtained at age 10-14 years and by serological tests obtained in adulthood did not influence the development of wheezing symptoms or atopic status in adulthood. However, early exposure to measles and family size may be associated with a lower risk of adult onset doctor diagnosed asthma.

Activation of host antitumoral responses by cationic lipid/DNA complexes.

A model of lipoplex-induced peritonitis was used to characterize the inflammatory response to cationic lipid:DNA lipoplexes with respect to activation of host antitumoral effector mechanisms. Three different cationic lipids were used in these studies: N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N-(1-[2,3-dioleoyloxylpropyl)-N,N,N-trimethylammonium chloride (DOTAP), and N-(1-[2,3-dioleyloxy]propyl)-N,N,N-trimethylammonium chloride (DOTMA). The DODAC and DOTMA lipoplexes exhibited similar transfection properties in vitro, whereas the DOTAP lipoplexes transfected quite poorly in all cell lines tested. Intraperitoneal injection of cationic lipoplexes into immunocompetent mice resulted in a profound infiltration of inflammatory cells, secretion of interferon-gamma, and increased natural killer activity within the peritoneal cavity. Both DODAC and DOTMA lipoplexes produced similar inflammatory responses, lasting at least 5 days. The inflammation induced by DOTAP lipoplexes peaked by day 3 and resolved to near-control levels by day 5. These data indicate that although cationic lipid DNA complexes may differ in their inflammatory properties, the natural killer activation and interferon-gamma secretion that follow lipoplex administration should provide a functional adjuvant for cancer gene therapies that benefit from immunostimulation.

Bronchial hyperresponsiveness and adult onset wheeze: the influence of atopy.

The commonly held belief that adult onset wheezing illness is primarily nonatopic in nature suggests that the role of atopy in the pathophysiology of bronchial hyperresponsiveness (BHR) in adult onset wheeze may be minimal. This study examined risk factors for BHR (BHR: provocative dose causing a 20% fall in forced expiratory volume in one second PD20 < or =16.38 micromol methacholine) among 82 subjects with adult onset wheeze and among 191 subjects who had never wheezed. Subjects were identified from a cohort of subjects aged 39-45 yrs who were known to have had no childhood wheeze and who were involved in a 30 yr follow-up survey. Risk factors for BHR were examined among all subjects with BHR and among subjects with BHR stratified according to whether or not they had ever wheezed. The prevalence of BHR was 40% (33/82) among the subjects with adult onset wheeze and 11% (21/191) among the subjects who had never wheezed. Lower baseline lung function (odds ratio (OR) = 0.94; 95% confidence interval (CI) = 0.92-0.97 per unit forced expiratory volume (FEV1)% predicted) and atopy (OR = 7.23; CI = 2.53-20.64 for all three measures of atopic compared to nonatopic) were associated with BHR, while smoking and family history showed no statistically significant relation to BHR. This pattern was also apparent in analyses stratified by symptom status. A family history of atopy increased the risk that BHR was accompanied by wheezing symptoms (OR = 4.75; CI = 1.53-14.72 for more than one affected relative compared to no affected relatives). These findings suggest that atopy is associated with bronchial hyperresponsiveness in adults known to have had no childhood wheeze. A familial factor reflecting genetic influences and/or shared environmental factors may influence whether bronchial hyperresponsiveness is associated with symptoms.

Mutational analysis of the DNA mismatch repair gene hMLH1 in myeloid leukaemias.

Mutations of the DNA mismatch repair (MMR) gene hMLH1 have recently been linked to the development of some hereditary and sporadic cancers which frequently display widespread microsatellite instability (MSI). Conflicting results regarding the extent of MSI in myeloid leukaemias prompted us to perform mutational analysis of all 19 exons of the hMLH1 gene by polymerase chain reaction-single-stranded conformation polymorphism (PCR-SSCP) and sequence analysis in a total of 133 patients with acute and chronic myeloid leukaemia. Apart from one exonic and one intronic polymorphism, no mutations were detected in any of the samples indicating that the major MMR gene hMLH1 is not involved in the pathogenesis or progression of myeloid malignancies.

Antioxidant intake and adult-onset wheeze: a case-control study. Aberdeen WHEASE Study Group.

An increase in prevalence of wheezing illness in the UK has coincided with a reduction in the consumption of natural antioxidants, which may modulate the lung's response to oxidant stress, limiting the expression of airway inflammation and respiratory symptoms. The hypothesis that intakes and plasma levels of natural antioxidants would be determinants of adult-onset wheezing illness was tested. A nested case-control study was conducted in 94 cases with adult-onset wheeze and 203 controls aged 39-45 yrs identified in a 30-yr follow-up survey. Antioxidant intake was measured by a food frequency questionnaire, and plasma and red cell measurements of antioxidant status were obtained. Outcome measures were onset of wheeze since age 15 yrs (ever wheeze) and wheeze occurring in the past 12 months (current wheeze). After adjusting for the effects of smoking, socioeconomic status, atopy, family history of atopic disease and total energy intake, intakes of vitamin E (odds ratio (OR) = 4.02 for low compared to high tertile of intake) and plasma levels of ascorbate (OR = 0.98 per unit) and alpha-tocopherol:triglyceride ratio (OR = 0.34 per log(e) unit) were inversely related to adult-onset wheeze. In analyses stratified by social class and smoking, intakes of vitamin C and E and plasma levels of ascorbate and alpha-tocopherol:triglyceride ratio were inversely related to current wheeze in the manual social class and among current smokers. No independent associations of vitamin A, beta-carotene or total plasma antioxidant capacity were found. The results support the hypothesis that deficiencies of vitamins C and E are associated with wheezing symptoms. Smokers in the manual social class are particularly susceptible to these effects.

Family size, childhood infections and atopic diseases. The Aberdeen WHEASE Group.

BACKGROUND: This study addresses the causes of the increases in childhood asthma and allergic disease. On the basis of an observed inverse relationship between family size and allergic disease or atopy, it has been proposed that a fall in common childhood infections may have been responsible for the rise in asthma. This study was undertaken to investigate the relationships between family size and reported allergic disease and to test the hypothesis that an inverse relationship between the two is a consequence of childhood infections. METHODS: Data had been obtained in a 1964 cross sectional survey of a random sample of Aberdeen schoolchildren aged between 10 and 14 in that year. Records of the presence or absence of asthma, eczema, or hay fever at the time of the survey and a history of measles, pertussis, varicella, rubella, and mumps before and after the age of three years were available for 2111 subjects. RESULTS: The risks of hay fever (odds ratio 0.2, 95% CI 0.1 to 0.8) and eczema (OR 0.3, CI 0.1 to 0.7) were inversely related to having had three or more older siblings, whilst the risk of asthma (OR 0.4, CI 0.1 to 0.9) was inversely related to having had three or more younger siblings. Increasing total numbers of siblings showed a significant trend in protection against both eczema and hay fever. A weak protective effect against asthma was found for measles after the age of three (OR 0.5, CI 0.3 to 0.9) and slight increases in the risk of eczema were associated with having had rubella or pertussis and of asthma with having had varicella. The number of infections before the age of three was associated with a significant trend in the odds ratios towards increased risk of asthma (p = 0.025). There were significant trends in the odds ratios towards greater risk of eczema and hay fever with increasing exposure to rubella, mumps, and varicella. These relations between infection and atopic diseases were independent of the potential confounding factors age, sex, father's social class, and total number of siblings. CONCLUSIONS: These data add to the accumulating evidence that membership of a large sibship confers some protection against atopic disease. This does not appear to be explained by the common childhood infections which show conflicting relationships with atopic disease, in that measles may have some protective effect against asthma but the more infections a child has had, the more likely he or she is to have atopic disease. The explanation of the sibship effect is likely to lie elsewhere and the fall in common childhood infections is unlikely to explain the rise in atopic disease.

[Snake bites in dogs]. / Schlangenbisse bei Hunden.

Altogether 11 cases of stings in dogs are reported. In at least nine cases, common vipers (Vipera berus) were identified to be the causes. The most frequently observed symptoms were haemorrhagic and oedematous inflammations of the area of the wounds, furthermore haemolysis and intense leukocytosis with relative and absolute neutrophilia and in some cases shock symptoms. One dog died 17 days after the accident from multiple organ insufficiencies, all the other dogs improved. As a therapy, bandage of the injured limb, quieting of the dog, snake antitoxin, corticosteroids, antihistaminics, antibiotics, and symptomatic therapy depending on the clinical signs are recommended.

Risk factors for adult onset wheeze: a case control study.

Risk factors associated with adult onset wheeze were examined in a case control study of subjects aged 39-45 yr derived from a community cohort of 2,056 asymptomatic children originally studied in 1964. Participants included 102 cases with adult onset wheeze (since age 15) and 217 controls with no wheeze. Logistic regression analysis was used to determine independent risk factors for wheeze among all cases and three subgroups: doctor diagnosed asthma (n = 24), wheeze with chronic cough and phlegm (n = 31), and other wheeze (n = 47). The risk of adult onset wheeze among all cases increased with low socioeconomic status (relative risk [RR] 2.36), current smoking (RR 2.01), positive atopic status (RR 3.28), and positive family history of atopic disease (RR 5.49). Gender was not related to the risk of wheezing. The pattern of significant independent risk factors differed between the subgroups of cases. Socioeconomic status was associated with cough and phlegm and other wheeze. Smoking habit was only related to cough and phlegm. Atopy was associated with doctor diagnosed asthma and cough and phlegm. Family history of atopic disease was related to all subgroups, suggesting that despite apparent heterogeneity in diagnostic labeling, concurrent symptoms, and other risk factors, the different forms of adult onset wheeze may share a common allergic basis.

Measuring health-related quality of life outcomes in women with endometriosis--results of the Gynaecology Audit Project in Scotland.

The clinical management of endometriosis was addressed within the recent Gynaecology Audit Project in Scotland. The impact of endometriosis and its treatment on women's health-related quality of life was examined using a condition-specific measure and a general measure, the Short Form 36 health survey (SF-36). Postal questionnaires containing the health-related quality of life measures were sent to 273 women at diagnosis and six months later. The measurement properties, including the reliability, validity and responsiveness, of the measures were examined. The condition-specific questions and the SF-36 had a high level of reliability. The validity of the condition-specific scores was demonstrated by their high correlation with the SF-36 which is a well-validated measure. Furthermore, the condition-specific scores were related to clinicians' assessment of disease severity and the need for further treatment. At the six month follow-up, changes in scores conformed to expected hypotheses, demonstrating the responsiveness of both measures. As a general measure, the SF-36 appeared to reflect the effects of both the condition of interest (i.e. endometriosis) and other conditions affecting health at the time of measurement (i.e. treatment side effects). The condition-specific measure was more responsive than the SF-36 to the changes in pain symptoms which resulted from active treatment. A condition-specific questionnaire, together with a general measure such as the SF-36 health survey, can provide a reliable, valid and responsive package of measures for assessing health-related quality of life in women with endometriosis. Such measures should be used alongside clinical measures of outcome to assess the effectiveness of different treatment strategies for endometriosis. A similar approach combining general and specific instruments would be useful in medical audits of other conditions.

Using economics alongside medical audit. A case study of the management of endometriosis.

During 1993, the diagnosis and initial management of endometriosis in women was identified as an important topic to be addressed within the Gynaecology Audit Project in Scotland (GAPS). This paper documents the experience of using the medical audit data collected by this exercise to estimate the costs and outcomes associated with alternative treatments for endometriosis associated infertility. Its aim is not to draw firm policy conclusions but rather to demonstrate the economic methods and to highlight difficulties in the application of economics to audit. The average cost of medical management was found to be significantly higher than the average cost of expectant management because of the high costs of the drugs involved. There was little difference in clinical and health outcomes observed between those women managed expectantly and those women managed medically. The results of the economic assessment add cautious support to the growing volume of clinical evidence indicating expectant management as the first line treatment of choice although further research is required to substantiate these results in a larger sample of women presenting with this condition.