bluuebunny/crossref_metadata_2025 huggingface dataset:
The morphological and spectral phenotype of apoptosis in HeLa cells varies following exposure to UV‐C and the addition of inhibitors of ICE and CPP32
Authors: Miller M. L. , Andringa A. , Elliott J. , Conwell Ii K. , Dixon K. , Carty M. P.
Published: December 2004

Numerous extra‐ and intracellular factors, including UV radiation, can initiate a programme of cell death by apoptosis. While apoptosis is commonly defined morphologically, the relationships between morphology and molecular events are not well established. To investigate these relationships in HeLa cells, eight morphometric criteria for cell proliferation and damage and 10 criteria for apoptotic phenotype were examined using light microscopy, and corroborated by ultrastructure and spectral imaging. They were identified (1) during a time course after irradiation with 0, 10 or 30 J/m2 UV‐C; (2) after separation of apoptotic from normal cells on a Percoll gradient; and (3) after irradiation with UV‐C plus perturbation of the apoptotic pathway by treatment with inhibitors of two caspases, ICE and CPP32.

The number of cells in apoptosis increased in a dose‐dependent manner after UV‐C treatment. Centrifugation of irradiated cells on a Percoll gradient increased the collection of apoptotic cells tenfold. The stereotypical apoptotic phenotype, in which cells have deep cytoplasmic blebbing and highly condensed DNA, comprised only a few percent of all apoptoses, and was rarely seen in groups receiving caspase inhibitors. The most common apoptotic phenotype was a rounded cell with large spherical nucleolus and associated DNA. After treatment with UV‐C plus inhibitors the apoptotic index was decreased by about 30% compared to UV‐C radiation alone. These apoptotic cells had dark spherical cytoplasm with small blebs, greatly increased numbers of cytoplasmic ribosomes, abundant nucleolar material with a large separate granular component, and chromatin condensed at the nuclear membrane. Using the technique of spectral imaging, it was found that the spectrum obtained from the granular component of the nucleolus, which was elevated in apoptotic cells treated with UV‐C plus inhibitors, was similar to the dense accumulation of ribosomes in the apoptotic cytoplasm. The data indicate that spectral imaging may be a useful tool for identifying and characterizing variations in the apoptotic process, and that the caspase inhibitors used here do not completely abolish UV‐C induced apoptosis, but rather alter its incidence and progression.
Genistein and erbstatin inhibition of normal mammary epithelial cell proliferation is associated with EGF‐receptor down‐regulation
Authors: McIntyre B. S. , Sylvester P. W.
Published: December 2004

Epidermal growth factor (EGF) is a potent mitogen for normal mouse mammary epithelial cells grown in primary culture. EGF activation of the EGF‐receptor (EGF‐R) induces intrinsic tyrosine kinase activity which results in EGF‐R autophosphorylation and tyrosine phosphorylation of other intracellular substrates involved in EGF‐R signal transduction. Genistein and erbstatin are anticancer agents which have been shown to be potent tyrosine kinase inhibitors. However, the effects of these compounds in modulating EGF‐dependent normal mammary epithelial cell proliferation is presently unknown. Therefore, studies were conducted to determine the effects of genistein and erbstatin on EGF‐dependent proliferation, and EGF‐R levels and autophosphorylation in normal mouse mammary epithelial cells grown in primary culture and maintained in serum‐free media. Chronic treatment with 6.25–100 μM genistein or 1–16 μM erbstatin significantly decreased EGF‐dependent mammary epithelial cell proliferation in a dose‐responsive manner. However, the highest doses of genistein (100 μM) and erbstatin (16 μM) were found to be cytotoxic. Additional studies showed that acute treatment with 6.25–400 μM genistein did not affect EGF‐R levels or EGF‐induced EGF‐R autophosphorylation, while acute treatment with 1–64 μM erbstatin caused a slight reduction in EGF‐R levels, but had no effect on EGF‐dependent EGF‐R autophosphorylation in these cells. In contrast, chronic treatment with similar doses of genistein or erbstatin resulted in a large dose‐responsive decrease in EGF‐R levels, and a corresponding decrease in total cellular EGF‐R autophosphorylation intensity. These results demonstrate that the inhibitory effects of chronic genistein and erbstatin treatment on EGF‐dependent mammary epithelial cell proliferation is not due to a direct inhibition of EGF‐R tyrosine kinase activity, but results primarily from a down‐regulation in EGF‐R levels and subsequent decrease in mammary epithelial cell mitogenic‐responsiveness to EGF stimulation.
Mammalian cells are not synchronized in G1‐phase by starvation or inhibition: considerations of the fundamental concept of G1‐phase synchronization
Authors: Cooper S.
Published: December 2004

Synchronization of mammalian cells by starvation‐refeeding or by inhibition‐release are among the most commonly used techniques for division cycle analysis. An alternative analysis—in the form of a Gedanken or thought experiment—is presented, casting doubt on the utility of this synchronization method. Arresting cell growth produces a culture where all cells contain a G1 amount of DNA. However, these cells are not arrested at a particular point in the G1‐phase. Analysis of ‘G1 arrested cells’ suggests that, upon resumption of growth, the cells are not synchronized.
Organizational support for evidence‐based practice within child and family social work: a collaborative study
Authors: Barratt Moira
Published: December 2004

Research in Practice works in collaboration with over 50 English local authorities and voluntary childcare organizations, to explore new and dynamic ways to increase the use of quality evidence to improve services to children and families. One Research in Practice initiative was a two‐year collaborative project involving the social services departments of six local authorities. This paper reports on how the views of more than 100 professional staff involved with the provision of services to children and families have been gathered to offer insight into how evidence‐based practice can be supported or frustrated in social care organizations. The findings suggest considerable uncertainty about the nature of evidence in social care and its validity in relation to decision‐making, policy and planning. Mechanisms essential for the dissemination, implementation and adoption of research messages are underdeveloped and tensions exist around the explicit use of research evidence within reports and reviews. Many practitioners and teams may be excluded from making decisions based on the best available research evidence through lack of access to internet resources and adequate information dissemination mechanisms. The paper concludes that there remain considerable areas for further debate if evidence‐based practice is to become a reality in work with children and families.
Who manages the care of students with Attention Deficit Hyperactivity Disorder (ADHD) in higher education?
Authors: Baverstock A. C. , Finlay F.
Published: December 2004

Objective  To identify who provides medical support to students with Attention Deficit Hyperactivity Disorder (ADHD) while in higher education.

Methods  A questionnaire pack was sent to 50 undergraduate student health centres attached to higher education colleges/universities.

Results  Eighty‐two per cent of practices returned questionnaires. Forty‐nine per cent had undergraduate students with ADHD, of those 76% were on methylphenidate. Fifty‐two per cent saw only their general practitioner (GP) for follow up and the rest were jointly managed by GP with: psychiatrists, paediatricians, psychologists or physicians. Eighty‐seven per cent of GPs had not attended recent courses or training on ADHD.

Conclusion  Those caring for undergraduate students with ADHD are still largely unfamiliar with the condition. Guidelines need to be drawn up to establish handover from paediatric to adult care.
The effects of work on the growth of Jordanian boys
Authors: Hawamdeh H. , Spencer N.
Published: December 2004

Objective  To examine the effects of work on growth among boys aged 10–16 years in Jordan.

Study design  Cross‐sectional health survey comparing working and non‐working boys.

Main outcomes  Height for age z‐score; weight for age z‐score.

Methods  One hundred and thirty‐five working and 405 non‐working boys aged 10–16 years were studied in the Jordanian areas of Irbid, Jarash and North Jordan Valley. The boys and their mothers were interviewed and data collected on work status, child's smoking status, and family socio‐economic characteristics including per capita income, family size, maternal and paternal education, area of the house in square metres and expenditure on household durables. Height and weight were obtained at the time of interview. Linear regression models were fitted on height for age z‐score and weight for age z‐score.

Results  In bivariate analysis, child's work status (r = −0.221), household per capita income (r = 0.104), family size (r = −0.102), house m2 per capita (r = 0.090) and monthly expenditure on durables (r = 0.086) were significantly correlated with height z‐score. Work status (P < 0.0001) and household m2 per capita (P = 0.002) were retained in the regression model fitted on height z‐score which explained 9.8% of the variance. The model fitted on weight z‐score explained 6.5% of the variance and work status (P < 0.0001), household per capita income (P = 0.041) were retained.

Conclusions  The results of this study suggest that, independent of a range of socio‐economic factors, work has a negative effect on child growth. Given the extent of child labour internationally these findings have profound implications for global child well‐being and for global social and economic policy.
Child health profiles: the viewpoints of adults
Authors: Hill M. , Morton P.
Published: December 2004

The child health profile  The teenage years have been identified as an important time to influence attitudes and behaviour likely to affect current and future health. The child health profile was developed partly to provide appropriate health information to young people aged 10 and older, but was also intended to recognize their right to assume greater responsibility for their health records. It was seen as an extension of the popular parent held record for younger children.

Methods  A Scottish child health profile was introduced through schools in three health board areas. Data about usage and views were obtained from children and relevant adults by means of questionnaires. In this paper, the perspectives of parents, health professionals and teachers are reported.

Findings  The majority of the adults were keen on the idea of the profile in principle. However, most parents knew little about it, even though all their children had been issued with one. Teachers had little involvement with the profile and the majority of health professionals had some doubts about its application. The adults indicated that a small number of children made effective use of the profile, but information from both adults and children indicated that it was seldom discussed during consultations with health professionals, few of whom routinely asked to see it.

Conclusions  The paper outlines implications for the provision of health information and records for this age group.
Behavioural problems, social competence and self‐concept in siblings of children with autism
Authors: Verté S. , Roeyers H. , Buysse A.
Published: December 2004

Introduction  This study investigated the psychological adjustment of siblings of children with high‐functioning autism (HFA) in comparison with siblings of normally developing children in the domain of behavioural problems, social competence and self‐concept.

Method   Twenty‐nine siblings of children with HFA and 29 siblings of children without a disorder participated in the study. Standardized, written questionnaires were used.

Results   Siblings of children with HFA, especially brothers and sisters between 6 and 11 years old, had more behavioural problems than siblings of the control group. Sisters of children with HFA ascribed higher social competence to themselves. Sisters of children with HFA between 12 and 16 years old had a more positive self‐concept. In both groups siblings with a more negative self‐concept had less social skills and siblings with a more positive self‐concept scored better in the social domain. Finally, in accordance with the control group, the perception of the siblings’ social competence of parents of children with HFA broadly matched the perception siblings had of themselves.

Discussion  Overall, siblings of children with HFA are not more susceptible to adaptation problems than siblings of children without a disorder. Implications for practice and further research are discussed.
Current practice in the management of Attention Deficit Disorder with Hyperactivity (ADHD)
Authors: Parr J. R. , Ward A. , Inman S.
Published: December 2004

Background  The management of Attention Deficit Disorder with Hyperactivity (ADHD) is controversial and guidance published by the National Institute for Clinical Excellence (NICE) suggests a multidisciplinary approach.

Methods  We evaluated the patterns of referral and techniques used in the diagnosis and management of ADHD in Wirral Health Authority by studying the case notes of 391 children diagnosed between 1994 and August 2000.

Results  The majority of cases were referred from Primary Care and the detected prevalence was 8/1000. Boys were diagnosed more frequently than girls (12 : 1) and the mean age at diagnosis was 8.7 years; girls were more likely to have been diagnosed prior to age 8. The use of behaviour rating questionnaires and school visits as an aid to diagnosis increased during the study period. Management by Child Psychiatry and Community Paediatric services was similar with stimulant medication prescribed in 91% of cases; Child Psychiatry services were more likely to give advice on behaviour management than Paediatric services. Twelve per cent of cases were managed by both services, or in joint Outpatient clinics. Outpatient follow‐up was most likely to occur at six monthly intervals and required 1312 appointments per year.

Conclusions  This study shows that the management of children with ADHD in Wirral Health Authority is compatible with NICE guidance. Attention Deficit Disorder with Hyperactivity is a significant burden on clinical services and long‐term planning is required.
Current Literature: 3
Authors: Reading Richard
Published: December 2004

Objective  To identify which clinical situations are the most difficult to manage for general paediatricians in Victoria, Australia.

Methods   Self‐administered questionnaires were sent to general paediatricians in Victoria. They were asked their opinions regarding what were the most difficult and the most dangerous clinical situations with which they deal.

Results   The response rate was 64% (63 out of 98 questionnaires sent). The general paediatricians surveyed believed that behavioural, developmental and psychosocial conditions were the most difficult to deal with; conduct disorder was the most nominated clinical category (26% of respondents). The ‘dangerous’ cases nominated were predominately traditional medical cases. The most commonly nominated category was sepsis and shock (21% of respondents). The most difficult and dangerous clinical situations overall for general paediatricians in Victoria appear to be in the areas of sepsis, child protection, paediatric and neonatal resuscitation, depression and suicide, raised intracranial pressure, intravenous fluid management, and communication with parents and adolescents.

Conclusions   The present survey provides useful information to help with training‐programme design and it gives trainee paediatricians an idea of what experienced paediatricians find difficult. Severe behavioural, family and social difficulties, as well as neonatal and childhood resuscitation, severe sepsis, raised intracranial pressure, and intravenous fluid management were the clinical situations most frequently described as difficult.
Parental history of atopic disease and concentration of cord blood IgE
Authors: JOHNSON C. C. , OWNBY D. R. , PETERSON E. L.
Published: December 2004

Background
A family history of atopy, and cord blood immunoglobulin E concentration, have been shown to be predictors of atopic disease in children. Several studies have suggested that parental atopy may be related to newborn immunoglobulin E. Objective The purpose of our analysis was to evaluate whether parental history of allergic disease was associated with cord blood immunoglobulin E concentration. Methods The study subjects were from a defined population of 777 newborns delivered between 1987 and 1989. The mothers of these children completed a questionnaire during pregnancy concerning themselves and the child's father, including parental history of physician diagnosis of allergic diseases (allergies, hay fever and asthma). Total immunoglobulin E levels were quantitated in cord blood samples with an enzyme‐hnked immunoassay.

Results
Median cord blood immunoglobulin E concentration was higher among infants whose mothers had a history of atopic disease, particularly for those with a history of asthma (P<0.022) and allergen immunotherapy (P<0.016) vs infants whose mothers had no history of any atopic disease. Comparing all babies with a maternal history of asthma, to babies where neither parent had a history of any atopic disease, the median cord blood immunoglobulin E was significantly higher (0.36IU/mL vs 0.21 IU/mL; P<0.009). This association was found only among female infants (0.49IU/mL vs 0.20 IU/mL; P<0.001).

Conclusion
Maternal, but not paternal, history of atopic disease was associated with an elevated immunoglobulin E among newborns. For maternal asthma, this association was only evident in infant girls.
Maternal fish oil supplementation in pregnancy reduces interleukin‐13 levels in cord blood of infants at high risk of atopy
Authors: Dunstan J. A. , Mori T. A. , Barden A. , Beilin L. J. , Taylor A. L. , Holt P. G. , Prescott S. L.
Published: December 2004

Background and objectives The epidemiological association between higher dietary n‐3 polyunsaturated fatty acids (PUFA) and lower prevalence of asthma, has led to interest in the role of early dietary modification in allergic disease prevention. In this study we examined the effects of maternal n‐3 (PUFA)‐rich fish oil supplementation on cord blood (CB) IgE and cytokine levels in neonates at risk of developing allergic disease.

Methods In a randomized double‐blind, placebo‐controlled trial, 83 atopic pregnant women received either fish oil capsules (n = 40) containing 3.7 g n‐3 PUFA/day or placebo capsules (n = 43) from 20 weeks gestation until delivery. CB cytokine levels (IL‐4, IL‐5, IL‐6, IL‐10, IL‐12, IL‐13, TNF‐α and IFN‐γ) and total IgE levels were measured and compared between the two groups. Fatty acid composition of red cell membranes was analysed by gas chromatography and the relationships among PUFA, cytokine and IgE levels were examined.

Results Maternal fish oil supplementation resulted in a significant increase in n‐3 PUFA levels (P < 0.001) in neonatal erythrocyte membranes. Neonates whose mothers had fish oil supplementation had significantly lower plasma IL‐13 (P < 0.05) compared to the control group. There was also a significant inverse relationship between levels of n‐3 PUFA in neonatal cell membranes and plasma IL‐13. There was no difference in levels of IgE and the other cytokines measured.

Conclusions This study provides preliminary evidence that increasing neonatal n‐3 PUFA levels with maternal dietary supplementation can achieve subtle modification of neonatal cytokine levels. Further assessment of immune function and clinical follow‐up of these infants will help determine if there are any significant effects on postnatal immune development and expression of allergic disease.
Mattress encasings and mite allergen levels in the Prevention and Incidence of Asthma and Mite Allergy study
Authors: Van Strien R. T. , Koopman L. P. , Kerkhof M. , Oldenwening M. , De Jongste J. C. , Gerritsen J. , Neijens H. J. , Aalberse R. C. , Smit H. A. , Brunekreef B.
Published: December 2004

Background Reduction of allergen exposure from birth may reduce sensitization and subsequent allergic disease.

Objective To measure the influence of mite allergen‐impermeable mattress encasings and cotton placebo encasings on the amount of dust and mite allergen in beds.

Methods A total of 810 children with allergic mothers took part in the Prevention and Incidence of Asthma and Mite Allergy (PIAMA) study. Allergen‐impermeable and placebo mattress encasings were applied to the childrens' and the parents' beds before birth. Dust samples were taken from the beds of children and their parents before birth and 3 and 12 months after birth. Extracts of dust samples were analysed for mite allergens (Der p 1 and Der f 1).

Results Active mattress encasings were significantly more effective in reducing dust and mite allergen levels than placebo encasings. Mite allergen levels were low in general and the treatment effect was modest. Twelve months after birth, mattresses with active mattress encasings had about half the amount of Der 1 (Der p 1 + Der f 1)/m2, compared to mattresses with placebo encasings, for the child's and the parental mattress.

Conclusion This study shows that mite‐impermeable mattress encasings have a significant but modest effect on dust and mite allergen levels of mattresses with low initial mite allergen levels, compared to placebo.
The economic burden of antibiotic treatment of penicillin‐allergic patients in internal medicine wards of a general tertiary care hospital
Authors: Sade K. , Holtzer I. , Levo Y. , Kivity S.
Published: December 2004

Background Penicillin allergy poses a major problem in the management of infectious diseases.

Objective We estimated the costs and usage of antibiotic treatment of ‘penicillin‐allergic’ patients in comparison to non‐allergic patients in a tertiary care hospital.

Materials and methods The study was based on the records of 118 randomly chosen in‐hospital patients labelled as being ‘allergic to penicillin’ and who were treated with antibiotics. The antibiotic selection and cost of the patients with alleged penicillin allergy were compared to 118 matched patients without an antibiotic allergy (controls).

Results During in‐hospital treatment, the mean antibiotic cost for penicillin‐allergic patients was 63% higher than the cost for the controls. In addition, there was a 38% higher cost of the recommended anti‐microbial treatment regimen to be followed upon discharge by the former compared to the latter.

Conclusions Penicillin‐allergic patients were more likely to receive broader spectrum antibiotics compared to the non‐allergic ones. Since many of the patients who are labelled as being ‘allergic to penicillin’ are, in fact, not allergic to it, inaccurate reporting of penicillin allergies may have costly economic and epidemiologic repercussions in addition to more toxic effects which can occur when choosing alternative drugs in case of penicillin allergy.
Systemic reactions to immunotherapy: influence of composition and manufacturer
Authors: Gastaminza G. , Algorta J. , Audicana M. , Etxenagusia M. , Fernández E. , Muñoz D.
Published: December 2004

Background Although immunotherapy clearly demonstrated the benefit of reducing allergic symptoms, it has the drawback of adverse events, mainly systemic reactions that could be very inconvenient for patients and even life‐threatening.

Objective The aim of the present study was to assess the incidence of systemic reactions to immunotherapy in a large number of patients, and its potential relationship with the characteristics of therapy, such as allergen composition or manufacturing laboratory.

Methods This study analysed the administration of specific immunotherapy during a period of 5 years, involving 1212 patients affected by respiratory hypersensitivity or hymenoptera venom anaphylaxis. Commercial extracts were supplied by five different laboratories. All the patients were attended at an out‐clinic immunotherapy unit by the same experienced staff. Immunotherapy was given following a conventional schedule, modified according the usual recommendations.

Results A total of 250 adverse reactions have been recorded, resulting in a frequency of 0.84% over the total number of injections. Seventy‐nine of them (32%) were systemic reactions (0.27% SR/injection). The 79 systemic reactions were observed in 60 patients (5% of the patients). The frequency of systemic reactions was significantly lower (P < 0.01) on the group of mites than on the other groups. The frequency of systemic reactions varies according to the manufacturing laboratory. In the case of mite extracts, although one of the laboratories had a lower frequency of adverse systemic reactions, it did not reach the level of statistical significance. However, in relation to pollen extracts, preparations of one of the manufacturers had a significantly lower frequency of systemic reactions. Concerning the time of occurrence, 27% of systemic reactions were delayed, thus they appeared at least 30 min after the vaccine injection, most of them due to pollen extracts.

Conclusion This is a preliminary study to evaluate the factors that could facilitate the appearance of systemic reactions demonstrating that not only the composition but also the manufacturer is connected to systemic reactions. Although further studies are needed to clearly establish the influence of manufacturer on frequency and time of appearance of systemic reactions, it seems necessary to reach a wide consensus on allergen extract standardization methods.
Cellular responses to cytomegalovirus in immunosuppressed patients: circulating CD8+ T cells recognizing CMVpp65 are present but display functional impairment
Authors: ENGSTRAND M , LIDEHÄLL A K , TÖTTERMAN T H , HERRMAN B , ERIKSSON B-M , KORSGREN O
Published: December 2004

The availability of tetrameric complexes of HLA class I molecules folded with immunodominant peptides makes it possible to utilize flow cytometry for rapid and highly specific visualization of virus specific CD8+ T cells. An alternate technique is to incubate whole blood with specific antigens and to subsequently detect and characterize responding T cells (e.g. by performing intracellular staining of interferon-gamma). By using an HLA-A2 tetramer construct folded with the same immunodominant CMV-peptide as that used for peptide pulsing, we monitored both the presence and functional capacity of CMV-specific CD8+ T cells. In addition T cell activation was assayed by determination of CD38 and CD69 expression. Twelve organ transplant patients and 31 healthy blood donors with latent CMV infection were investigated using CMV pp65 tetramer staining and intracellular staining of interferon-gamma after CMV pp65 peptide pulsing or CMV lysate pulsing. CMV-specific T cells were detected in similar absolute numbers as well as frequencies of T cells in the two groups investigated. However, the CMV-specific CD8+ T cells in immunosuppressed individuals showed a decreased functional response to the CMV-peptide, as evidenced by reduced interferon-gamma production when compared to healthy blood donors (19%; 42%, P < 0·005). In addition, CD38 expression was markedly higher in immunosuppressed patients compared to healthy blood donors (24%; 6%, P < 0·005). In a case report we demonstrate that reactivation of CMV can occur in an immunosuppressed patient with high number of CMV-specific T cells, but without functional capacity. Hence, these findings reflect impaired activation of cytotoxic T cells controlling latent CMV infection in immunosuppressed patients.
High serum vascular endothelial growth factor correlates with disease activity of spondylarthropathies
Authors: DROUART M , SAAS P , BILLOT M , CEDOZ J-P , TIBERGHIEN P , WENDLING D , TOUSSIROT É
Published: December 2004

Angiogenesis is involved in chronic inflammatory joint diseases such as rheumatoid arthritis (RA). Vascular endothelial growth factor (VEGF) plays a crucial role in angiogenesis. The spondylarthropathies (SpA) are characterized by enthesitis and synovitis, in which blood vessels participate. The objective of this study was to investigate serum VEGF levels and their potential associations with disease activity markers for SpA. Sera were collected from 105 patients with SpA (72 with ankylosing spondylitis (AS), four with psoriatic arthritis (PsA), six with reactive arthritis (ReA), eight with enteropathic arthropathy and 15 with undifferentiated SpA), 50 patients with rheumatoid arthritis (RA) and 64 healthy controls. Disease activity in SpA patients was assessed using the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and laboratory parameters of inflammation [erythrocyte sedimentation rate (ESR) and C-reactive protein level (CRP)]. Serum VEGF levels were significantly higher in SpA patients (316·4 ± 215·6 pg/ml) and RA patients (405·2 ± 366·5) than in controls (217·3 ± 145·2) (P = 0·003). In SpA patients, serum VEGF levels correlated with disease activity indices (BASDAI: r = 0·22, P = 0·04; ESR: r = 0·3, P = 0·003; and CRP: r = 0·23, P = 0·02). Serum VEGF levels were not associated with presence of extra-articular manifestations or syndesmophytes or with the grade of sacroiliitis. These results suggest that VEGF and therefore angiogenesis may play a role in SpA pathogenesis and may serve as a disease activity marker in SpAs.
Keratinocyte growth factor (KGF) decreases ICAM-1 and VCAM-1 cell expression on bronchial epithelial cells
Authors: JUST N , TILLIE-LEBLOND I , GUERY B P H , FOURNEAU C , TONNEL A-B , GOSSET P
Published: December 2004

Activation of leucocytes during airway inflammatory reaction involves adhesion to bronchial epithelial cells (BEC), a process implicating specific interactions between glycoproteins with epithelial cell surface proteins, mainly intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). In this study, the effect of keratinocyte growth factor (KGF), a growth factor involved in pulmonary epithelium repair, was evaluated on adhesion molecule expression with BEAS-2B cells and BEC and granulocyte adherence to BEAS-2B. The modulation by KGF of membrane and mRNA expression of ICAM-1 and VCAM-1 was studied on confluent cells stimulated or not with tumour necrosis factor-α (TNF) (200 UI/ml) or TNF and interleukin (IL)-4 (50 UI/ml and 10 ng/ml). Levels of soluble-(s)ICAM-1 and sVCAM-1 were measured by ELISA. Although moderately, KGF significantly decreased membrane ICAM-1 expression in unstimulated BEAS-2B cells (24% inhibition at 100 ng/ml) or in TNF- or TNF + IL-4-stimulated cells (22·5 and 18·7% inhibition, respectively). Treatment with KGF tended to decrease VCAM-1 expression in TNF- and TNF + IL-4-stimulated BEAS-2B (P = n.s. and P < 0·05, 14 and 15% inhibition, respectively). In primary culture of BEC, adhesion molecule expression was also reduced. ICAM-1 and VCAM-1 mRNA expression were also inhibited by KGF. Levels of sICAM-1 and sVCAM-1 were not significantly increased in supernatants from KGF-treated cells (30% and 24% increase at 100 ng/ml, respectively) compared to controls. Moreover, KGF decreased by 31% the adherence of neutrophils to TNF-activated BEAS-2B. In conclusion, KGF decreases ICAM-1 and VCAM-1 expression and neutrophil adherence in BEC. These suggest its involvement in the resolution of the inflammatory reaction.
Augmented production of proinflammatory cytokines and accelerated allotransplantation reactions in heroin-treated mice
Authors: HOLÁŇ V , ZAJÍCOVÁ A , KRULOVÁ M , BLAHOUTOVÁ V , WILCZEK H
Published: December 2004

Heroin treatment or abusive drug addiction influences many physiological functions, including the reactions of the immune system. Although suppression of various manifestations of the immune system after heroin (or morphine) administration has been reported, we show here that production of proinflammatory cytokines and nitric oxide (NO) was enhanced and allotransplantation reactions were accelerated significantly in heroin-treated recipients. Mice were treated by a subcutaneous administration of heroin (diacetylmorphine) given in one or repeated daily doses. The ability of spleen cells from treated mice to respond in vitro to alloantigens and to produce IL-2, IL-4, IL-10 and IFN-γ, and the production of IL-1β, IL-12 and NO by peritoneal macrophages, were tested. Within 2 h after heroin administration, proliferative responses to alloantigens and the production of IL-1β, IFN-γ, IL-12 and NO were enhanced significantly. In contrast, the production of anti-inflammatory cytokines IL-4 and IL-10 was at the same time rather decreased. As a consequence, skin allografts in heroin-treated mice were rejected more promptly than in untreated or vehicle-treated recipients. Similarly, the growth of allogeneic tumours induced by high doses of tumour cells was suppressed significantly in heroin-treated mice. The enhancing effects of heroin on the production of proinflammatory cytokines were antagonized by naltrexone, a specific inhibitor of classic opioid receptors. These results show that heroin treatment augments production of proinflammatory cytokines and accelerates allotransplantation reactions. The observations thus illustrate the complexity of the effects of heroin on the immune system and should be taken into account during medical treatment of opiate addicts and in the use of morphine to decrease pain in various clinical situations.
Increased circulating interleukin-12 (IL-12) p40 in pulmonary sarcoidosis
Authors: SHIGEHARA K , SHIJUBO N , OHMICHI M , KAMIGUCHI K , TAKAHASHI R , MORITA-ICHIMURA S , OHCHI T , TATSUNO T , HIRAGA Y , ABE S , SATO N
Published: December 2004

In sarcoidosis, a T helper 1 (Th1) response is an essential event and the up-regulation of interleukin-12 (IL-12) has been detected in affected disease sites. In order to investigate the clinical usefulness of circulating IL-12, we measured the serum concentrations of IL-12 by ELISA and performed immunohistochemistry using specific MoAbs for IL-12 in the lungs and scalene lymph nodes of patients with sarcoidosis. The serum concentration of IL-12 p40 was detectable in all 45 patients with pulmonary sarcoidosis and 18 normal controls, whereas that of IL-12 p70 was undetectable. The serum concentrations of IL-12 p40 in pulmonary sarcoidosis were significantly higher than those of the normal controls, especially in cases with abnormal intrathoracic findings detected by chest roentogenogram. The serum concentrations of interferon-γ (IFN-γ) also increased compared with those of normal controls and there was a significant positive correlation between the serum concentrations of IL-12 p40 and IFN-γ. Furthermore, serum angiotensin-converting enzyme (ACE) and lysozyme, which are known to be useful markers for disease activity in sarcoidosis, correlated well with the serum concentrations of IL-12 p40. The positive 67Ga scan group (for lung field) had significantly elevated serum IL-12 p40 levels compared with those of the negative group. No bioactivity of IL-12 p70 was detected in three sarcoid cases sera by using the IL-12 responsive cell line. Finally, the immunohistochemical approach revealed that IL-12 p40 was expressed in the epithelioid cells and macrophages of sarcoid lungs and lymph nodes. We concluded that the production of IL-12 p40 was far greater in the sera and we have demonstrated this to be a useful clinical marker for disease activity and the Th1 response in pulmonary sarcoidosis.
Role of incidental and/or cured intestinal parasitic infections on profile of CD4+ and CD8+ T cell subsets and activation status in HIV-1 infected and uninfected adult Ethiopians
Authors: KASSU A , TSEGAYE A , WOLDAY D , PETROS B , AKLILU M , SANDERS E J , FONTANET A L , VAN BAARLE D , HAMANN D , DE WIT T F RINKE
Published: December 2004

Intestinal parasitic infections have been suggested to cause persistent immune activation leading to an unbalanced immune state. Such a state has been proposed to be a major factor in the pathogenesis of AIDS in an African context. The present study investigated the effect of incidental parasitic infection and treatment on the profile of T cell differentiation and activation markers on CD4+ and CD8+ T cells from HIV-1 infected and uninfected adult Ethiopians. Cryopreserved PBMCs from 64 subjects (41 HIV-negative and 23 HIV-positive) with follow-up visits at 6-monthly intervals were used to compare the effect of incidental intestinal parasites and their treatment upon T cell subset profiles and activation status. The samples were stained with antibodies to various T cell differentiation and activation markers allowing naive, memory, effector, memory/effector, activated and resting CD4+ and CD8+ T cell subsets to be quantified by triple-colour FACScan. Incidental intestinal parasitic infections resulted in a significant increase in memory CD4+ T cell numbers both in HIV-negative and HIV-positive subjects (P < 0·05). There was also a significant increase in the percentage of CD8+ HLA-DR+ T cells (P < 0·05) in HIV-positive subjects co-infected with parasites. In HIV-negative subjects, a significant decline in activated cells and a significant increase in resting CD8+ T cells (P < 0·05) was observed after treatment for parasites. These data suggest that intestinal parasitic infections could result in the alteration of T cell subset counts and also in the up-regulation of T cell activation markers in

peripheral blood. Treatment of parasitic infections showed a tendency to reduce the activation suggesting that, together with other community based intervention strategies, such treatment could be used to down-regulate immune activation and hence protect the host from being easily attacked by HIV.
CpG ODN enhances uptake of bacteria by mouse macrophages
Authors: UTAISINCHAROEN P , KESPICHAYAWATTANA W , ANUNTAGOOL N , CHAISURIYA P , PICHYANGKUL S , KRIEG A M , SIRISINHA S
Published: December 2004

Unmethylated CpG motif in synthetic oligodeoxynucleotide (CpG ODN) or bacterial DNA is well recognized for its role in innate immunity, including enhancing production of NO and cytokines by macrophages. In the present study, we demonstrated the effect of CpG ODN on the phagocytic uptake of bacteria by macrophages. Flow cytometric analysis of mouse macrophages (RAW 264·7) incubated with fluorescein isothiocyanate (FITC)-labelled Burkholderia pseudomallei, Salmonella enterica serovar Typhi or Escherichia coli showed that CpG ODN increased the uptake of these bacteria by mouse macrophages. The enhancement of bacterial uptake by CpG ODN was concentration-dependent. The increase of bacterial uptake by CpG ODN-activated macrophages shown above is consistent with the result of bacteria internalization study using a standard antibiotic protection assay. There was also an increase in the rate and degree of multi-nucleated giant cell formation, phenomena which have been shown previously to be unique when the cells were infected with B. pseudomallei. These observations may provide significant insights for future investigation into host cell–pathogen interaction.
Glomerular deposition of C1q and anti-C1q antibodies in mice following injection of antimouse C1q antibodies
Authors: TROUW L A , SEELEN M A , DUIJS J M G J , BENEDIKTSSON H , VAN KOOTEN C , DAHA M R
Published: December 2004

Anti-C1q autoantibodies are present in the serum of patients with different autoimmune diseases such as systemic lupus erythematosus (SLE). The occurrence of these autoantibodies correlates with renal involvement. In the present study we examined whether injection of rabbit antimouse C1q antibodies in mice leads to deposition in kidneys. Injection of healthy mice with a single dose of rabbit IgG antimouse C1q antibodies resulted in deposition of both C1q and IgG anti-C1q in glomeruli. The pattern of deposition observed in the glomeruli of mice injected with antimouse C1q antibodies both at 24 h and 2 weeks was both glomerular basement membrane (GBM)-associated and mesangial. Injection of control IgG did not have a detectable effect on circulating C1q levels, and no deposition of either C1q or rabbit IgG was seen at 24 h. The deposition of rabbit antimouse C1q and C1q in glomeruli resulted in complement activation, as assessed by C3 deposition, and influx of leucocytes associated with albuminuria in some, but not all mice. In none of the control mice was albuminuria observed. This report is the first to show that anti-C1q antibodies deposit in the healthy glomerulus together with autologous C1q. This deposition is stable for at least 2 weeks, causes complement activation, leucocyte influx and can lead to mild albuminuria.
Anti-cytokine autoantibodies in autoimmunity: preponderance of neutralizing autoantibodies against interferon-alpha, interferon-omega and interleukin-12 in patients with thymoma and/or myasthenia gravis
Authors: MEAGER A , WADHWA M , DILGER P , BIRD C , THORPE R , NEWSOM-DAVIS J , WILLCOX N
Published: December 2004

We have screened for spontaneous anticytokine autoantibodies in patients with infections, neoplasms and autoimmune diseases, because of their increasingly reported co-occurrence. We tested for both binding and neutralizing autoantibodies to a range of human cytokines, including interleukin-1alpha (IL-1α), IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-18, interferon-alpha2 (IFN-α2), IFN-ω, IFN-β, IFN-γ, tumour necrosis factor alpha (TNF-α), transforming growth factor beta-1 (TGF-β1) and granulocyte-macrophage colony stimulating factor (GM-CSF), in plasmas or sera. With two notable exceptions described below, we found only occasional, mostly low-titre, non-neutralizing antibodies, mainly to GM-CSF; also to IL-10 in pemphigoid. Strikingly, however, high-titre, mainly IgG, autoantibodies to IFN-α2, IFN-ω and IL-12 were common at diagnosis in patients with late-onset myasthenia gravis (LOMG+), thymoma (T) but no MG (TMG–) and especially with both thymoma and MG together (TMG+). The antibodies recognized other closely related type I IFN-α subtypes, but rarely the distantly related type I IFN-β, and never (detectably) the unrelated type II IFN-γ. Antibodies to IL-12 showed a similar distribution to those against IFN-α2, although prevalences were slightly lower; correlations between individual titres against each were so modest that they appear to be entirely different specificities. Neither showed any obvious correlations with clinical parameters including thymoma histology and HLA type, but they did increase sharply if the tumours recurred. These antibodies neutralized their respective cytokine in bioassays in vitro; although they persisted for years severe infections were surprisingly uncommon, despite the immunosuppressive therapy also used in most cases. These findings must hold valuable clues to autoimmunizing mechanisms in paraneoplastic autoimmunity.
Altered CD8+ T cell responses to selected Epstein–Barr virus immunodominant epitopes in patients with multiple sclerosis
Authors: HÖLLSBERG P , HANSEN H J , HAAHR S
Published: December 2004

An increased frequency of antiviral CD8+ T cells is seen in chronic viral infections. During herpes virus infections the expanded CD8+ T cells are thought to control the reactivation of the latent infection. Because multiple sclerosis (MS), a presumed autoimmune disease of the central nervous system, has been associated with a late Epstein–Barr virus (EBV) infection, we wished to examine whether the CD8+ T cell response to EBV epitopes differed between MS patients and healthy controls. Here we report an increased frequency of CD8+ T cells responding to EBV epitopes from nuclear antigen 3 A (HLA-A2/CLG) and latent membrane protein 2 (HLA-B7/RPP) in MS patients. Noticeably, the altered CD8+ T cell response occurred to some but not all EBV epitopes and did not reach the high level seen during acute infection. The responses towards two immunodominant epitopes from human cytomegalovirus (HCMV) were similar in MS patients and normal controls. Together, our data demonstrate the presence of an increased frequency of CD8+ T cells reacting with two epitopes from EBV in patients with MS. The altered response to only two of the tested EBV epitopes would be consistent with the presence of cross-reactive epitopes.
T lymphocyte phenotypic profile in lung segments affected by cavitary and non-cavitary tuberculosis
Authors: MAZZARELLA G , BIANCO A , PERNA F , D'AURIA D , GRELLA E , MOSCARIELLO E , SANDUZZI A
Published: December 2004

Clinical manifestations of pulmonary tuberculosis (TB) may depend on a complex interaction between the host and the pathogen. Clinical outcomes of pulmonary tuberculosis are variable, ranging from asymptomatic lifelong infection to parenchymal lung destruction, resulting in cavitary lesions. To investigate the hypothesis that local cellular immune response may affect presentation and outcome in tuberculosis, we performed bronchoalveolar lavage (BAL) in lung segments affected by cavitary and non-cavitary tuberculosis. We then correlated the type of cellular response at the level of the involved lung segments with clinical evolution in terms of cavity formation. We found alveolar lymphocytosis in patients with both cavitary and non-cavitary pulmonary tuberculosis, with increased CD4+ lymphocytes in patients with non-cavitary pulmonary tuberculosis. A predominant Th1 immune response has been observed in non-cavitary patients, while cavitary involved segments exhibit the presence of Th2 lymphocyte subsets. These data, while confirming the importance of Th1-type CD4+ cells and IFN-γ in effective cellular immunity in active pulmonary tuberculosis, also suggest that the presence of Th2 lymphocytes may contribute to tissue necrosis phenomena associated with cavitary evolution of pulmonary tuberculosis. Our observations indicate the importance of the type of local immune response at the site of disease in the development of different clinical characteristics and outcome in pulmonary tuberculosis.
Mannan-binding lectin and hepatitis C infection
Authors: KILPATRICK D C , DELAHOOKE T E S , KOCH C , TURNER M L , HAYES P C
Published: December 2004

Japanese patients with chronic hepatitis C infection unresponsive to treatment with interferon possessed genotypes disproportionately conferring low mannan-binding lectin (MBL) concentrations. Our aims were to confirm or refute this finding in European patients at the MBL protein level, and to investigate whether a low circulating concentration of MBL might influence susceptibility to, or disease progression from, hepatitis C viral infection. Serum samples obtained from 180 hepatitis C patients and 566 blood donors were assayed for MBL. MBL concentrations were related to disease characteristics retrieved from patients’ records. MBL concentrations were higher in hepatitis C patients (median 2·5 µg/ml versus 1·3; P < 0·0001) and the proportion of patients with very low (MBL-deficient) concentrations was similar to that of the healthy controls. There were no significant associations between patients with low serum MBL and the disease features studied, including response to antiviral therapy. Therefore, low circulating MBL does not increase susceptibility to hepatitis C infection, and MBL concentration does not have a major influence on the course of the disease or the response to antiviral therapy. MBL replacement therapy would therefore not be indicated for chronic hepatitis C patients who failed to respond fully to treatment with interferon and ribavirin.