Monobody adapter for functional antibody display on nanoparticles for adaptable targeted delivery applications.

Vascular endothelial cells (ECs) play a central role in the pathophysiology of many diseases. The use of targeted nanoparticles (NPs) to deliver therapeutics to ECs could dramatically improve efficacy by providing elevated and sustained intracellular drug levels. However, achieving sufficient levels of NP targeting in human settings remains elusive. Here, we overcome this barrier by engineering a monobody adapter that presents antibodies on the NP surface in a manner that fully preserves their antigen-binding function. This system improves targeting efficacy in cultured ECs under flow by >1000-fold over conventional antibody immobilization using amine coupling and enables robust delivery of NPs to the ECs of human kidneys undergoing ex vivo perfusion, a clinical setting used for organ transplant. Our monobody adapter also enables a simple plug-and-play capacity that facilitates the evaluation of a diverse array of targeted NPs. This technology has the potential to simplify and possibly accelerate both the development and clinical translation of EC-targeted nanomedicines.

Investigating changes in anger and aggression in offenders with high levels of psychopathic traits attending the Chromis violence reduction programme.

BACKGROUND: Chromis was accredited by the Correction Services Accreditation Panel in 2005 as an intervention designed to reduce violence in offenders whose level or combination of psychopathic traits disrupts their ability to engage in treatment and change. It runs as part of the regime in the dangerous and severe personality disorder unit in HM Prison Frankland (Westgate). A multiple case study investigation into changes over time in participants is currently underway, part of which is reported here. AIMS: This paper reports on information relating to changes in anger and aggression in Chromis completers. METHODS: Change in psychometrics and observed incidents of verbal and physical aggression are considered for five case study participants who have completed Chromis and progressed from Westgate to a different location. RESULTS: Findings suggest that cases experienced a reduction in self-reported anger, and expected incidents of physical aggression but had higher than expected levels of verbal aggression after leaving Westgate. CONCLUSIONS: These findings offer cautious optimism for the effectiveness of Chromis, although methodological limitations must also be considered. Findings may be seen as positive indicators of Chromis, or at least the approach to working with these offenders across Westgate, in reducing violence. IMPLICATIONS FOR PRACTICE: Findings support the continued delivery and evaluation of Chromis. There may be benefit in exploring ways to further understand and address verbal aggression in participants.

Immunohistochemistry in the evaluation of neovascularization in tumor xenografts.

Angiogenesis, or neovascularization, is known to play an important role in the neoplastic progression leading to metastasis. CD31 or Factor VIII-related antigen (F VIII RAg) immunohistochemistry is widely used in experimental studies for quantifying tumor neovascularization in immunocompromised animal models implanted with transformed human cell lines. Quantification, however, can be affected by variations in the methodology used to measure vascularization including antibody selection, antigen retrieval (AR) pretreatment, and evaluation techniques. To examine this further, we investigated the microvessel density (MVD) and the intensity of microvascular staining among five different human tumor xenografts and a mouse syngeneic tumor using anti-CD31 and F VIII RAg immunohistochemical staining. Different AR methods also were evaluated. Maximal retrieval of CD31 was achieved using 0.5 M Tris (pH 10) buffer, while maximum retrieval of F VIII RAg was achieved using 0.05% pepsin treatment of tissue sections. For each optimized retrieval condition, anti-CD31 highlighted small vessels better than F VIII RAg. Furthermore, the MVD of CD31 was significantly greater than that of F VIII RAg decorated vessels (p<0.001). The choice of antibody and AR method has a significant affect on immunohistochemical findings when studying angiogenesis. One also must use caution when comparing studies in the literature that use different techniques and reagents.

Localization of electrogenic Na/bicarbonate cotransporter NBCe1 variants in rat brain.

The activity of HCO(3)(-) transporters contributes to the acid-base environment of the nervous system. In the present study, we used in situ hybridization, immunoblotting, immunohistochemistry, and immunogold electron microscopy to localize electrogenic Na/bicarbonate cotransporter NBCe1 splice variants (-A, -B, and -C) in rat brain. The in situ hybridization data are consistent with NBCe1-B and -C, but not -A, being the predominant NBCe1 variants in brain, particularly in the cerebellum, hippocampus, piriform cortex, and olfactory bulb. An antisense probe to the B and C variants strongly labeled granule neurons in the dentate gyrus of the hippocampus, and cells in the granule layer and Purkinje layer (e.g. Bergmann glia) of the cerebellum. Weaker labeling was observed in the pyramidal layer of the hippocampus and in astrocytes throughout the brain. Similar, but weaker labeling was obtained with an antisense probe to the A and B variants. In immunoblot studies, antibodies to the A and B variants (alphaA/B) and C variant (alphaC) labeled approximately 130-kDa proteins in various brain regions. From immunohistochemistry data, both alphaA/B and alphaC exhibited diffuse labeling throughout brain, but alphaA/B labeling was more intracellular and punctate. Based on co-localization studies with antibodies to neuronal or astrocytic markers, alphaA/B labeled neurons in the pyramidal layer and dentate gyrus of the hippocampus, as well as cortex. alphaC labeled glia surrounding neurons (and possibly neurons) in the neuropil of the Purkinje cell layer of the cerebellum, the pyramidal cell layer and dentate gyrus of the hippocampus, and the cortex. According to electron microscopy data from the cerebellum, alphaA/B primarily labeled neurons intracellularly and alphaC labeled astrocytes at the plasma membrane. In summary, the B and C variants are the predominant NBCe1 variants in rat brain and exhibit different localization profiles.

Targeting cellular memory to reprogram the epigenome, restore potential, and improve somatic cell nuclear transfer.

Successful cloning by somatic cell nuclear transfer (SCNT) is thought to require reprogramming of a somatic nucleus to a state of restored totipotentiality [Dean, W., Santos, F., Reik, W., 2003. Epigenetic programming in early mammalian development and following somatic cell nuclear transfer. Semin. Cell. Dev. Biol. 14, 93-100; Jouneau, A., Renard, J.P., 2003. Reprogramming in nuclear transfer. Curr. Opin. Genet. Dev. 13, 486-491; ]. Though SCNT-induced reprogramming is reminiscent of the reprogramming that occurs after fertilization, reprogramming a differentiated nucleus to an embryonic state is delayed and incomplete in comparison (for review, see ). This is likely due to the existence of an epigenetic-based cellular memory, or program, that serves to regulate global patterns of gene expression, and is the basis of a genome defense mechanism that silences viruses and transposons. The mechanisms of this memory include CpG methylation and modification of histones. Recent evidence by Feng et al. [Feng, Y.-Q., Desprat, R., Fu, H., Olivier, E., Lin, C.M., Lobell, A., Gowda, S.N., Aladjem, M.I., Bouhasira, E.E., 2006. DNA methylation supports intrinsic epigenetic memory in mammalian cells. PLOS Genet. 2, 0461-0470], using a transgenic experimental system, indicates that these marks may be acquired in more than one order and thus, silent heterochromatic structure can be initiated by either methylation of CpG dinucleotides or by histone modifications. In this system, however, CpG methylation appears to differ from histone modifications because it bestows a persistent epigenetic, or cellular, memory. In other words, CpG methylation can independently confer cellular memory, whereas histone modifications appear to be limited in this capacity. Therefore, in the context of genomic reprogramming induced by SCNT, efficient demethylation is likely a key (if not the only) rate-limiting step to improving the efficiency and outcomes of SCNT cloning. This review discusses the possibility of targeting cellular memory, and in particular inducing demethylation of a somatic nucleus prior to nuclear transfer, to enable reprogramming events typically carried out by oocyte factors and thereby improve developmental competence of SCNT-reconstructed embryos. Several recent published reviews of SCNT, cellular reprogramming and genomic demethylation served as valuable sources for the authors and are recommended as supplemental reading. These include the following: Bird, A., 2002. DNA methylation patterns and epigenetic memory. Gen. Dev. 16, 6-21; Grafi, G., 2004. How cells dedifferentiate: a lesson from plants. Dev. Biol. 268, 1-6; Latham, K.E., 2005. Early and delayed aspects of nuclear reprogramming during cloning. Biol. Cell 97, 119-132; Lyko, F., Brown, R., 2005. DNA methyltransferase inhibitors and the development of epigenetic cancer therapies. J.Natl. Cancer Inst. 97, 1498-1506; Morgan, H.D., Santos, F., Green, K., Dean, W., Reik, W., 2005. Epigenetic reprogramming in mammals. Hum. Mol. Gen. 14, R47-R58; Szyf, M., 2005. DNA methylation and demethylation as targets for anticancer therapy. Biochemistry 70, 533-549; Buszczak, M., Spradling, A.C., 2006. Searching chromatin for stem cell identity. Cell 125, 233-236; Gurdon, J.B., 2006. From nuclear transfer to nuclear reprogramming: the reversal of cell differentiation. Annu. Rev. Cell. Dev. Biol. 22, 1-22; Yoo, C.B., Jones, P.A., 2006. Epigenetic therapy of cancer: past, present and future. Nat. Rev. 5, 37-50.

A comparison of survival models for assessing risk of racehorse fatality.

Survival analysis was used to assess risk factors for fatal injuries on UK race courses. This allowed assessment of variation due to temporal horse-level effects, including previous racing intensity and historical distribution of race types, as well as race-level factors. Comparisons were made between measuring survival time as number of days and as number of races to injury from the first race. Two related models were presented for time as number of races to injury: a Cox regression model fitted using partial likelihood, with the Efron approximation to handling ties, and a discrete-time logit model fitted using maximum likelihood. The latter approach had the advantages of being computationally more efficient and enabling the testing of different functional forms for the dependence of hazard on time. Retrospective data were available from all race starts on the 59 courses in Britain from 1990 to the end of 1999, as analysed by . The analysis was conducted on the data for the 47,424 horses that had started racing in the UK: 538,895 starts with 1,228 fatal injuries. Horses starting racing abroad were excluded, but some included horses would have raced abroad at some stage during their racing career. The results for the selected models were broadly consistent with each other and with previously published studies. Steeplechase and hurdle races had a higher risk of fatal injury than flat races (relative hazards 1.5 and 1.7, respectively). Risk increased with the firmness of surface, age and race distance (reaching a plateau at 20 furlongs) and decreased with previous racing intensity (reaching a plateau after seven races run in the last 12 months). Horses running their first race of a new type were also found to be at higher risk (relative hazard 1.5). The main difference between the models for time as number of days and number of races concerned the role of age: age at race was identified as the more important factor in the latter model, whereas, age at first race was more significant in the former model.

Evidence for peroxynitrite-mediated modifications to p53 in human gliomas: possible functional consequences.

Based on previous findings of increased nitric oxide synthase (NOS) expression in human gliomas (4), we hypothesized that peroxynitrite, a highly reactive metabolite of nitric oxide (NO) and superoxide (O(*-)(2)), might be increased in these tumors in vivo. Here we demonstrate that nitrotyrosine (a footprint of peroxynitrite protein modification) is present in human malignant gliomas. Furthermore, we show that p53, a key tumor suppressor protein, has evidence of peroxynitrite-mediated modifications in gliomas in vivo. Experiments in vitro demonstrate that peroxynitrite treatment of recombinant wild-type p53 at physiological concentrations results in formation of higher molecular weight aggregates, tyrosine nitration, and loss of specific DNA binding. Peroxynitrite treatment of human glioma cell lysates similarly resulted in selective tyrosine nitration of p53 and was also associated with loss of p53 DNA binding ability. These data indicate that tyrosine nitration of proteins occurs in human gliomas in vivo, that p53 may be a target of peroxynitrite in these tumors, and that physiological concentrations of peroxynitrite can result in a loss of p53 DNA binding ability in vitro. These findings raise the possibility that peroxynitrite may contribute to loss of wild-type p53 functional activity in gliomas by posttranslational protein modifications.

Racehorse injuries, clinical problems and fatalities recorded on British racecourses from flat racing and National Hunt racing during 1996, 1997 and 1998.

For improvements to the safety and welfare of racehorses to be possible, it is essential to have access to basic descriptive information about the veterinary incidents encountered during horseracing. A 3 year surveillance study (1996-1998) was conducted by The Jockey Club into racing injuries, other postrace clinical problems and fatalities from all 59 British racecourses (mainland Britain only) to identify risk factors. During the survey there were 222,993 racing starts: 106,897 starts in flat races on turf (47.9%), 26,519 starts in flat races on all-weather surfaces (11.9%), 30,932 starts in chases on turf (13.9%), 51,786 starts in hurdle races on turf (23.2%) and 6,859 starts in National Hunt flat races (3.1%). Information was recorded about age of horses, racing surfaces and clinical events observed or attended by a veterinary team of 2 clinicians and one veterinary surgeon employed by the racing authority. Of the 2358 clinical events reported (1.05% of all starts), 1937 involved the musculoskeletal system and 421 involved other body systems. Six hundred and fifty-seven incidents (0.29% of starts) resulted in death or euthanasia. Eighty-one percent of limb injury reports involved forelimbs and 46% involved flexor tendons/suspensory ligaments. Nonlimb problems included epistaxis (0.83/1000 starts), 'exhausted horse syndrome' (0.47/1000 starts) and paroxysmal atrial fibrillation (0.20/1000 starts). Incidents including fatalities per 1000 starts were 24.7 from chases, 19.45 from hurdle races, 8.46 from National Hunt flat races and 3.97 from flat races. The overall tendon injury was higher in chases than in hurdle races, even though age-specific rates of tendon injury were higher in hurdle races than in chases. The risk of injuries per start increased significantly with age, while softer racing surfaces were associated with fewer fatalities and injuries than firmer surfaces. The survey described in this paper has provided an up-to-date description of the fatal and non-fatal horseracing incidents under conditions on mainland Britain, enabling progress to be made towards improving the safety and welfare of racehorses.

Oxidative stress and heat shock stimulate RGS2 expression in 1321N1 astrocytoma cells.

RGS2, a regulators of G-protein signaling family member, regulates G-protein signaling and is itself controlled in part by regulated expression. We tested if cell stress regulates RGS2 expression in human astrocytoma 1321N1 cells. Treatment with H2O2 increased RGS2 mRNA levels time- and concentration-dependently, with 200 microM H2O2 causing an approximately eightfold increase after 2 h. Peroxynitrite and heat shock also increased RGS2 mRNA levels. H2O2-induced RGS2 expression was negatively regulated by phosphoinositide-3-kinase and extracellular signal-regulated kinases. H2O2 also concentration-dependently increased RGS2 protein levels, and the RGS2 appeared to be predominantly in the nucleus. These results demonstrate that RGS2 expression is up-regulated by cell stress.

HuR, a RNA stability factor, is expressed in malignant brain tumors and binds to adenine- and uridine-rich elements within the 3' untranslated regions of cytokine and angiogenic factor mRNAs.

Tumors of the central nervous system (CNS) often have sustained expression of labile genes, including angiogenic growth factors and immunosuppressive cytokines, which promote tumor progression. Stabilization of the RNA transcripts for these genes, such as vascular endothelial growth factor (VEGF), is an important molecular pathway for this up-regulation. HuR, a member of the Elav family of RNA-binding proteins, has been implicated in this pathway through its binding to adenine and uridine (AU)-rich stability elements (ARE) located in the 3' untranslated regions (3'-UTRs) of the mRNA. Whereas three of the Elav family members (Hel-N1, HuC, and HuD) are restricted to young and mature neurons, HuR is more broadly expressed, including proliferating cells of the developing CNS. Because RNA stabilization of labile genes may promote tumor growth, we analyzed and compared the expression pattern of HuR in 35 freshly resected and cultured CNS tumors to determine whether there was any correlation with tumor grade or histological type. We found that HuR mRNA was consistently expressed in all of the tumors, regardless of cell origin or degree of malignancy. Using a novel HuR-specific polyclonal antibody, we found that strong HuR protein expression was limited to high-grade malignancies (glioblastoma multiforme and medulloblastoma). Within the glioblastoma multiforme, prominent HuR expression was also detected in perinecrotic areas in which angiogenic growth factors are up-regulated. To further define its role as a potential RNA stabilizer, we analyzed whether HuR could bind to the stability motifs within the 3'-UTRs of cytokines and growth factors linked to brain tumor progression. We used a novel ELISA-based RNA binding assay and focused on the 3'-UTRs of angiogenic factors VEGF, COX-2, and (interleukin) IL-8 as well as the immunomodulating factors IL-6, transforming growth factor (TGF)-beta and tumor necrosis factor (TNF)-alpha as potential RNA ligands. Our results indicated overall a very high binding affinity to these RNA targets. A comparison of these ligands revealed a hierarchy of binding affinities with the angiogenic factors, and TGF-beta showing the highest (Kd of 1.8-3.4 nM), and TNF-alpha the lowest (Kd of 18.3 nM). The expression pattern of HuR, coupled with the RNA binding data, strongly suggests a role for this protein in the posttranscriptional regulation of these genes in CNS tumors.

Second messengers regulate RGS2 expression which is targeted to the nucleus.

Regulators of G-protein Signaling (RGS) proteins attenuate signaling activities of G proteins, and modulation of expression appears to be a primary mechanism for regulating RGS proteins. In human astrocytoma 1321N1 cells RGS2 expression was increased by activation of muscarinic receptors coupled to phosphoinositide signaling with carbachol, or by increased cyclic AMP production, demonstrating that both signaling systems can increase the expression of a RGS family member in a single cell type. Carbachol-stimulated increases in endogenous RGS2 protein levels appeared by immunocytochemical analysis to be largely confined to the nucleus, and this localization was confirmed by Western blot analysis which showed increased nuclear, but not cytosolic, RGS2 after carbachol treatment. Additionally, transiently expressed green fluorescent protein (GFP)-tagged, 6xHis-tagged, or unmodified RGS2 resulted in a predominant nuclear localization, as well as a distinct accumulation of RGS2 along the plasma membrane. The intranuclear localization of GFP-RGS2 was confirmed with confocal microscopy. Thus, RGS2 expression is rapidly and transiently increased by phosphoinositide signaling and by cyclic AMP, and endogenous and transfected RGS2 is largely, although not entirely, localized in the nucleus.

HIV+ patients have increased lymphocyte infiltrates in CIN lesions.

OBJECTIVE: The objective of our study was to analyze immunocyte infiltrates in CIN lesions from HIV+ patients to assess whether local immunosuppression, defined as a decrease in T cell infiltrates, could explain the aggressive nature of CIN in HIV-infected patients. MATERIALS AND METHODS: Cervical tissue was obtained from 6 HIV+ CIN patients, 6 HIV- CIN patients, who underwent LLETZ (large loop excision of the transformation zone) for CIN, and 17 normal patients who underwent hysterectomy for benign indications. The following cell surface markers were analyzed: CD20 (B cells), CD4 (T helper cells), and CD8 (T suppressor/cytotoxic cells). Each tissue section was visualized with a Leica microscope at 400x and the image was captured for analysis by Harmony Group image analysis software. RESULTS: A significantly higher number of lymphocytes (both B and T cells) was detected in the stroma of HIV+/CIN tissue compared to either HIV-/CIN or normal tissue. There was also a significant increase in CD8+ cells in the HIV+/CIN group compared to HIV-/CIN or normal tissue. There was a trend toward a decreased CD4+/CD8+ ratio in the HIV+/CIN compared to the other two groups; however, this did not reach statistical significance. CONCLUSIONS: This study indicates that HIV+/CIN cervical tissue has a greater number of tissue lymphocytes recruited to the neoplastic site compared to HIV- individuals. In addition, HIV+ patients may have a decreased CD4/CD8 ratio in locally infiltrating immunocytes in CIN lesions. The local immunomodulatory effects of HIV may be detectable early in infection and therefore may explain the aggressive nature of CIN in the HIV+ patient.

Hay fever, eczema, and wheeze: a nationwide UK study (ISAAC, international study of asthma and allergies in childhood).

OBJECTIVES: To describe the prevalence of atopic symptoms in children throughout the UK. METHOD: A questionnaire survey of 12-14 year olds throughout England, Wales, Scotland, and the Scottish Islands using the international study of asthma and allergies in childhood (ISAAC) protocol. RESULTS: A total of 27 507 (86%) children took part. Recent rhinoconjunctivitis was reported by 18.2%, with 6.2% reporting symptoms between March and September; 16.4% reported itchy flexural rash in the past 12 months. The prevalence of atopic symptoms was higher in girls and subjects born within the UK. The prevalence of severe wheeze was highest in subjects reporting perennial rhinoconjunctivitis, as opposed to summertime only symptoms. Winter rhinoconjunctivitis was associated with severe wheeze and severe flexural rash. One or more current symptoms were reported by 47.6% of all children and 4% reported all three symptoms. CONCLUSION: In general, geographical variations were small but the prevalence of symptoms was significantly higher in Scotland and northern England. The study demonstrates the importance of atopic diseases both in their own right and in association with asthma.

Are women more ready to consult than men? Gender differences in family practitioner consultation for common chronic conditions.

BACKGROUND: When consultations for all reasons are combined, women are seen to consult their general practitioners more than men through most of adult life. It is, therefore, often assumed that women are more likely to consult for every condition. OBJECTIVES: To examine whether women report being more likely to consult a general practitioner than men when taking account of the underlying condition and various aspects of the experience of the condition consulted for. METHODS: Home-based nurse-interviews with 852 people in early middle age (39 years) and 858 in late middle age (58 years) sampled from the general population in the West of Scotland. Detailed information about current chronic conditions included general practitioner consultation and reported experience of pain frequency, pain severity, limitation to normal activities and restricted activity in the previous four weeks. RESULTS: Women were no more likely than men to consult a general practitioner in the previous year when experiencing the five most common groups of conditions; in addition, women were no more likely than men to consult at a given level of severity for a given condition type, except in the case of one aspect of reported experience of mental health problems. CONCLUSIONS: The results argue against the most widely accepted explanation for gender differences in consulting, namely, that women are simply more likely to consult a general practitioner than men irrespective of underlying morbidity. Reasons for the higher rates of women consulting observed in general practice-based studies are discussed in relation to these data.

Respiratory symptoms and home environment in children: a national survey.

BACKGROUND: Respiratory diseases are common in childhood and may lead to chronic disease in adult life; environmental factors probably play an important part in their causation. METHODS: A survey of respiratory symptoms in children aged 12-14 years was conducted throughout Great Britain as part of the International Study of Asthma and Allergies in Childhood (ISAAC). Information was obtained on certain aspects of the home environment in order to assess their importance as risk factors. RESULTS: The response rate was 79.3%, and 25,393 children in 93 schools participated. In a multiple regression analysis, wheeze was reported more often in nonmetropolitan areas and in association with active smoking, passive smoking, the presence of a furry pet, bottled gas, paraffin, and other unusual heating fuels; small regional differences persisted. Current smoking, previous smoking, and passive smoking accounted for 10.4%, 6.8%, and 6.5%, respectively, of wheezing in the past 12 months, and furry pets accounted for 5.0%. Cough and phlegm were associated with active and passive smoking and with the miscellaneous fuels; similar associations were found for rhinitis, but were less consistent for rhinitis occurring in spring and summer. Gas cooking showed little association with respiratory symptoms. CONCLUSIONS: Passive as well as active smoking is an important cause of respiratory symptoms in adolescence. Pets seem to increase the risk of wheeze and rhinitis, and fumes from certain unusual heating fuels may have adverse effects. Home environment and geographical location have independent effects on the prevalence of respiratory symptoms.

Loss of bcl-2 expression in ductal carcinoma in situ of the breast relates to poor histological differentiation and to expression of p53 and c-erbB-2 proteins.

AIM: This study (1) investigates the incidence of bcl-2 protein expression in a series of 108 cases of ductal carcinoma in situ (DCIS), including 25 with early invasive carcinoma, and (2) evaluates the relationship of bcl-2 expression to the histological grade of DCIS and to the expression of oestrogen receptor (ER), c-erbB-2 and p53 proteins. METHODS AND RESULTS: The expression of bcl-2, oestrogen receptor (ER), c-erbB-2 and p53 proteins was determined immunohistochemically. Cases were regarded as positive for individual antibodies when at least 10% of the DCIS cells showed positive staining. DCIS was graded histologically as well (n = 9), intermediately (n = 24), or poorly differentiated (n = 75). bcl-2 expression was documented in 57 cases (53%) and was strongly associated with the histological grade of DCIS (P < 0.0001). All cases of well-differentiated DCIS were bcl-2 positive and loss of bcl-2 expression was almost exclusively confined to poorly differentiated DCIS lesions. bcl-2 expression was also closely associated with positive ER status (P < 0.0001). Forty-seven of 57 (82%) bcl-2 positive cases were ER positive while 49/51 (96%) bcl-2 negative cases were ER negative. There was a significant inverse correlation between bcl-2 expression and both p53 protein expression (P = 0.0004) and c-erbB-2 expression (P < 0.0001). Nineteen of 24 (79%) p53 positive cases and 38/45 (84%) c-erbB-2 positive cases showed loss of bcl-2. CONCLUSIONS: Loss of bcl-2 expression occurs in poorly differentiated DCIS and is related to negative ER status and to positive p53 and c-erbB-2 status. This pattern of bcl-2 expression and its association with other biological markers in DCIS is similar to that reported in invasive breast carcinoma.

Prevalence of asthma symptoms, diagnosis, and treatment in 12-14 year old children across Great Britain (international study of asthma and allergies in childhood, ISAAC UK)

OBJECTIVE: To investigate variations in the prevalence of self reported symptoms, diagnosis, and treatment of asthma in 12-14 year old children. DESIGN: Self completion questionnaire. SETTING: Great Britain. SUBJECTS: All pupils aged 12-14 years in a stratified cluster sample of 93 large mixed secondary schools in 1995. MAIN OUTCOME MEASURES: Self reported prevalence of symptoms, diagnosis, and treatment of asthma at four geographical levels. RESULTS: 27,507 questionnaires were completed (85.9% response rate). The national 12 month prevalence of any wheezing, speech limiting wheeze, four or more attacks of wheeze, and frequent night waking with wheeze was 33.3% (n = 9155), 8.8% (2427), 9.6% (2634), and 3.7% (1023) respectively. The prevalence of ever having had a diagnosis of asthma was 20.9% (5736). In total, 19.8% (5438/27,507) of pupils reported treatment with anti-asthma drugs in the past year, but, of pupils reporting frequent nocturnal wheeze in the past year, 33.8% (342/1012) had no diagnosis of asthma and 38.6% (395/1023) denied receiving inhaler therapy. The 12 month prevalence of wheeze was highest in Scotland (36.7%, 1633/4444), but in England and Wales there was no discernible north-south or east-west gradient. Wheeze prevalence was slightly higher in non-metropolitan areas (35.0%, 6155/17,605) than in metropolitan areas (30.3%, 3000/9902). The prevalence of self reported asthma diagnosis and inhaler use showed no discernible national, regional, north-south, or east-west geographical pattern but was higher in non-metropolitan areas. CONCLUSION: Prevalence of self reported symptoms, diagnosis, and treatment of asthma was high among 12-14 year olds throughout Great Britain with little geographical or urban-rural variation. Underdiagnosis and undertreatment were substantial.

Sibship size and self-reported inhalant allergy among adult women. ALSPAC Study Team.

BACKGROUND: An association of allergic sensitization with small families and low birth order has been described and attributed to a protective effect of early infection. The influence of like-sex and unlike-sex siblings has not been investigated, although the severity of viral infections may be greater if acquired from unlike-sex siblings. OBJECTIVE: To investigate the association of self-reported inhalant allergy with family composition. METHODS: Reports of allergy to grass, dust or cats by 11042 pregnant women recruited to a longitudinal study of pregnancy and childhood in Avon, UK, were analysed in relation to respondent's age, maternal age and sibship composition (older and younger brothers and sisters) by multiple logistic regression. RESULTS: The prevalence of self-reported inhalant allergy decreased with increasing numbers of brothers (test for trend: P < 0.0001), but was unrelated to the number of sisters. The unadjusted prevalences for subjects with none, one, two and three or more brothers were 26%, 23%, 20% and 17%, respectively. The corresponding prevalences for numbers of sisters were 23%, 24%, 22% and 23%. After adjustment for total sibship size, offspring of older mothers were more likely to report allergy (test for trend: P < 0.001), but there was no association with position in the sibship. CONCLUSION: Although it is not possible to determine whether brothers specifically, or unlike-sex siblings in general, are inversely associated with inhalant allergy, these findings are consistent with the hypothesis that patterns of sibling interaction within young families influence the risk of future aeroallergen sensitization.

Childhood antecedents of allergic sensitization in young British adults.

OBJECTIVE: This study was designed to investigate whether associations of self-reported hay fever with sibship size, birth order, infant feeding, and childhood socioeconomic status reflect variations in sensitization to common aeroallergens. METHODS: One thousand three hundred sixty-nine persons born throughout Britain in 1958 were followed up to age 34 to 35 years. The cohort included 1050 subjects with a history of asthma, wheezy bronchitis, wheezing, or pneumonia and 319 with no history of wheezing illness at ages 7, 11, 16, 23, or 33 years. Skin prick tests with extracts of mixed grass pollen, house dust mite (Der p 1), and cat fur were performed; and wheal diameters were measured. RESULTS: The prevalence of positive skin test results (> or = 3 mm wheal) was independently related (p < 0.01) to male sex, reduced numbers of older siblings (but not younger siblings), and higher socioeconomic status in childhood. Current cigarette smoking and maternal smoking during pregnancy were independently associated (p < 0.01) with a reduced prevalence of skin prick test positivity. No significant independent effects (p > 0.10) were found for adult social class, maternal age, birth weight, gestation, breast feeding, preschool nursery attendance, urban birthplace, or gas stove exposure. CONCLUSION: Factors related to small families and relative affluence in childhood promote atopic sensitization to a variety of aeroallergens in later life. These observations are consistent with the suggestion that early infection may protect against subsequent allergic disease.