bluuebunny/crossref_metadata_2025 huggingface dataset:
A new single nucleotide polymorphism in the chicken pituitary‐specific transcription factor (POU1F1) gene associated with growth rate
Authors: Jiang R. , Li J. , Qu L. , Li H. , Yang N.
Published: July 2004

The pituitary‐specific transcription factor (POU1F1) is a protein which binds to and transactivates promoters of growth hormone (GH), prolactin (PRL) and thyroid‐stimulating hormone chain (TSHB)‐encoding genes. Ten chicken populations (n = 662), including six Chinese indigenous breeds, White Leghorn, paternal/maternal lines of brown egg layer and a paternal line of broiler, were used to detect single nucleotide polymorphisms in the pituitary‐specific transcription factor gene (PIT1) by means of PCR‐SSCP. A nucleotide transversion from adenine (A) to thymidine (T) at position 980 of the open reading frame of the PIT1 cDNA (GenBank accession no. AF029892) was identified. This nucleotide transversion results in an alteration of codon 299 from AAC to ATC, which leads to a change from asparagine (Asn) to isoleucine (Ile) in the POU domain of POU1F1. The distribution of allele and genotype frequencies differed significantly between meat‐type chickens (higher frequencies of A and A/A) and layer‐type chickens (P < 0.01). Another experimental population with growth records was used to evaluate the relationship between this polymorphism and growth rate. The results revealed a positive relationship between genotype A/A and body weight at 8 weeks of age, indicating that the SNP in PIT1 gene is a potential molecular marker for early growth rate in chicken.
The genes encoding bovine SP‐A, SP‐D, MBL‐A, conglutinin, CL‐43 and CL‐46 form a distinct collectin locus on Bos taurus chromosome 28 (BTA28) at position q.1.8–1.9
Authors: Gjerstorff M. , Hansen S. , Jensen B. , Dueholm B. , Horn P. , Bendixen C. , Holmskov U.
Published: July 2004

Collectins are a group of C‐type lectins involved in the innate immune system, where they mediate and modulate clearance of pathogens. The health status of cattle is of major economical and ethical concern; therefore, the study of bovine collectins is of importance. The collectins conglutinin, CL‐43 and CL‐46 are only present in Bovidae and the characterization of their genes indicates that they are structural descendants of another collectin, lung surfactant protein D (SP‐D). In this study, we assembled BAC clones into a contig spanning 330–1150 kb, which includes the bovine genes encoding the collectins SP‐A (SFTPA), SP‐D (SFTPD), mannan‐binding lectin A (MBL1), CL‐43 (COLEC9), CL‐46 (COLEC13) and conglutinin (COLEC8). In the same contig, we also identified a gene that potentially encodes a novel conglutinin‐like collectin (COLEC14). The arrangement of STFPA, SFTPD and MBL1 is homologous to the organization found in humans and mice, whereas the Bovidae‐specific collectin genes, COLEC8, COLEC9 and COLEC13, extend from SFTPD. Proximal to the collectin locus at BTA28q1.8–1.9, and included in the contig, we found the microsatellite IDVGA8, which may be a valuable marker for tracking polymorphisms in the linked collectin genes.
Contributions of Portuguese cattle breeds to genetic diversity using marker‐estimated kinships
Authors: Mateus J. C. , Eding H. , Penedo M. C. T. , Rangel‐Figueiredo M. T.
Published: July 2004

The quantitative assessment of genetic diversity within and between populations is important for decision‐making in genetic conservation plans. In our study, we applied the livestock core set method to define the contribution of 15 cattle breeds, 11 of which are Portuguese indigenous cattle breeds, to genetic diversity. In livestock core set theory genetic diversity is defined as the maximum genetic variance that can be obtained in a random‐mating population that is bred from the populations present in that core set. Two methods to estimate marker‐estimated kinships to obtain the contributions to the core set were used in this study: the weighted log‐linear model (WLM) and the weighted log‐linear mixed model (WLMM). The breeds that contributed most to diversity in the core set were Holstein‐Friesian followed by the Portuguese Mertolenga and Cachena for both WLM and WLMM methods. The ranking of relative contributions of cattle breeds was maintained when we considered only the Portuguese cattle breeds. Furthermore, we were able to identify the marginal contributions and respective losses of diversity for each of the 11 Portuguese cattle breeds when we considered a subset of populations that are not threatened of being lost (the Safe set composed of the four exotic breeds present in this study). When WLM was used losses in genetic diversity ranged from 2.68 to 0.65% while the loss in founder genome equivalents ranged from 37.37 to 8.43% for Mertolenga and Brava de Lide breeds respectively. When WLMM was used losses in genetic diversity and founder genome equivalents were less extreme than for the WLM method, ranging from 1.27 to 0.69 and 26.8 to 12.99 respectively.
Antipsychotic polypharmacy in patients with schizophrenia: a multicentre comparative study in East Asia
Authors: Sim Kang , Su Alex , Fujii Senta , Yang Shu‐yu , Chong Mian‐Yoon , Ungvari Gabor S. , Si Tianmei , Chung Eun K. , Tsang Hin‐Yeung , Chan Yiong H. , Heckers Stephan , Shinfuku Naotaka , Tan Chay H.
Published: July 2004

Aims

Previous studies of the prescription patterns of psychotropic medications in patients with schizophrenia have highlighted a high rate of antipsychotic polypharmacy, but data in Asia are sparse. This study seeks to examine the prevalence of antipsychotic polypharmacy in patients with schizophrenia and compare the differences between patients receiving one vs. those receiving more than one antipsychotic.

Methods

Antipsychotic prescription for a sample of 2399 patients with schizophrenia from six countries and territories was evaluated. Daily doses of antipsychotic medications were converted to standard chlorpromazine equivalents (CPZ).

Results

Antipsychotic polypharmacy was found in 45.7% (n = 1097) of the patients with wide intercountry variations. Polypharmacy was associated with male gender [odds ratio (OR) 1.24, 95% confidence interval (CI) 1.06, 1.46, P < 0.01], advanced age (t = −7.81, d.f. = 2396, P < 0.001), psychiatric hospital setting (OR 1.34, 95% CI 1.11, 1.62) as well as higher daily CPZeq doses (411.47 vs. 983.10 CPZeq day−1, z = −25.94, P < 0.001), anticholinergic use (OR 3.17, 95% CI 2.65, 3.79, P < 0.001) and less use of an atypical antipsychotic drug (OR 0.83, 95% CI 0.71, 0.98, P < 0.05). On multivariate analysis, country, age and duration of illness were significantly associated with antipsychotic polypharmacy.

Conclusion

This study highlighted the wide intercountry variations of antipsychotic polypharmacy which are likely to be influenced by a complex combination of clinical, setting, cultural and personal practice factors, requiring more research.
A comparison of lung function methods for assessing dose–response effects of salbutamol
Authors: Houghton Catherine M. , Woodcock Ashley A. , Singh Dave
Published: July 2004

Pulmonary function methods which are able to detect small pharmacological effects may be useful for assessing the full dose–response curve of bronchodilatators. We compared the ability of impulse oscillometry (R5, R20, X5, RF), plethysmography (sGaw) and spirometry [forced expiratory volume in 1 s (FEV1), maximal mid expiratory flow rate (MMEF)] to measure the dose–response effects of salbutamol in 12 healthy subjects, 12 mild asthmatics (mean FEV1 96% predicted) and 12 moderate asthmatics (mean FEV1 63% predicted). The techniques were performed twice to assess variability. Then salbutamol 10, 20, 100, 200 and 800 µg was administered. The sensitivity of the methods were compared by determining the lowest dose that caused changes greater than variability. In healthy subjects significant changes (p ≤ 0.05) were observed only in FEV1 (4.1%) and MMEF (14.6%) at 100 µg and sGaw (25.6%) and R20 (8.3%) at 200 µg. In mild asthmatics significant changes were observed in sGaw (15.9%) at 10 µg, X5 (23%), RF (20.3%) and MMEF (15.7%) at 20 µg, R5 (13.9%) and R20 (9.4%) at 100 µg and FEV1 (7.1%) at 200 µg. All measurements except R20 demonstrated significant changes at 10 µg in moderate asthmatics. The most sensitive test for assessing bronchodilatation is different in healthy subjects and asthmatics, and varies with severity of airflow obstruction.
In vitro fertilization and childhood retinoblastoma
Authors: Bradbury Brian D. , Jick Hershel
Published: July 2004

Aims

To estimate the frequency of childhood retinoblastoma among children conceived by in vitro fertilization (IVF).

Methods

Using the United Kingdom‐based General Practice Research Database (GPRD), we identified all live births, cases of retinoblastoma and IVF procedures occurring between January 1989 and December 2001.

Results

We identified 0 cases of retinoblastoma among 176 children conceived by IVF (Risk = 0/176, one‐sided 97.5% CI 0, 0.02) and 24 cases of retinoblastoma diagnosed before age 5 years among 358 094 children not conceived by IVF (6.7 cases per 100 000 births [95% CI 4.5, 10]).

Conclusions

These data provide some reassurance that children born as a result of IVF are not at markedly increased risk of retinoblastoma.
In vitro blood distribution and plasma protein binding of the tyrosine kinase inhibitor imatinib and its active metabolite, CGP74588, in rat, mouse, dog, monkey, healthy humans and patients with acute lymphatic leukaemia
Authors: Kretz Olivier , Weiss H Markus , Schumacher Martin M , Gross Gerhard
Published: July 2004

Aims

To determine blood binding parameters of imatinib and its metabolite CGP74588 in humans and non‐human species.

Methods

The blood distribution and protein binding of imatinib and CGP74588 were determined in vitro using 14C labelled compounds.

Results

The mean fraction of imatinib in plasma (fp) was 45% in dog, 50% in mouse, 65% in rat, 70% in healthy humans and up to 92% in acute lymphatic leukaemia (AML) patients. Similarly, fp for CGP74588 was low in dog and monkey (30%), higher in rat, mouse and humans (70%) and highest in some AML patients (90%). The unbound fraction of imatinib and CGP74588 in plasma was lower in rat, mouse, healthy humans and AML patients (2.3–6.5% at concentrations ≤ 5000 ng ml−1) compared to monkey and dog (7.6–19%). Both compounds displayed high binding to human α1‐acid glycoprotein. AML patients had a reduced haematocrit and showed greatest variability in their blood binding parameters.

Conclusion

Imatinib and CGP74588 displayed very similar blood binding parameters within all species/groups investigated. The five species clustered into two distinct groups with rat, mouse and humans being clearly different from dog and monkey. For both compounds, higher protein binding was associated with a decreased partitioning into blood cells.
Plasma concentrations during naltrexone implant treatment of opiate‐dependent patients
Authors: Olsen Linda , Christophersen Asbjørg S. , Frogopsahl Grete , Waal Helge , Mørland Jørg
Published: July 2004

Aims

To evaluate individual variations in plasma concentrations over time in patients with naltrexone implants.

Methods

Ten opioid‐dependent patients received up to four implants. Plasma samples were collected regularly for the analyses of naltrexone and the metabolite β‐naltrexol.

Results

The median naltrexone Cmax was 12.3 (range 5.8–22.1) ng ml−1, the median Tmax was 1 day (range 3 h to 35 days), and the median length of time that plasma concentrations were above 1 ng ml−1 was 55 (range 30–80) days. Two patients reported heroin use without experiencing any effect. Tissue reactions were recorded in two patients after repeated implantation.

Conclusion

Marked individual and intraindividual variations in naltrexone concentrations were observed. Further studies should be performed to evaluate the need for therapeutic drug monitoring during naltrexone implant treatment.
Leflunomide in active rheumatoid arthritis: a prospective study in daily practice
Authors: Van Roon E. N. , Jansen T. L. TH. A. , Mourad L. , Houtman P. M. , Bruyn G. A. W. , Griep E. N. , Wilffert B. , Tobi H. , Brouwers J. R. B. J.
Published: July 2004

Aims  We prospectively studied the efficacy, incidence of adverse drug reactions and withdrawal from leflunomide in an outpatient population with rheumatoid arthritis in a setting of care‐as‐usual.

Methods  In this prospective case series study, a standard dataset was collected from outpatient medical records, including patient and disease characteristics, data on leflunomide use and adverse drug reactions.

Results  During the study period 136 rheumatoid arthritis patients started leflunomide. Median (range) follow‐up duration was 317 (11–911) days. Sixty‐five percent of patients experienced at least one adverse drug reaction related to leflunomide. During follow‐up 76 patients (56%) withdrew from leflunomide treatment, mainly because of adverse drug reactions (29%) or lack of efficacy (13%). The overall incidence density for withdrawal from leflunomide was 56.2 per 100 patient years. Complete data for calculating efficacy using a validated disease activity score on 28 joints (DAS28) was available for 48, 36, and 35% of patients at 2, 6, and 12 months follow‐up, respectively. Within a 12‐month period after start of leflunomide treatment 76% of the evaluable patients were classified as moderate or good responders according to the DAS28 response criteria.

Conclusions  In the setting of care‐as‐usual rheumatoid arthritis patients starting leflunomide frequently experienced adverse drug reactions. More than half of the patients withdrew from leflunomide treatment within 1 year of starting leflunomide treatment, mainly because of adverse drug reactions.
Robustness of chlorzoxazone as an in vivo measure of cytochrome P450 2E1 activity
Authors: Ernstgård Lena , Warholm Margareta , Johanson Gunnar
Published: July 2004

Aims

Chlorzoxazone is metabolized by cytochrome P450 2E1 (CYP2E1) to a single oxidized metabolite, 6‐hydroxychlorzoxazone. The aim of the study was to test the robustness of chlorzoxazone as an in vivo probe of CYP2E1 activity in humans, with emphasis on investigating short‐term and long‐term intra‐individual variabilities and effects of different doses of the drug. In addition, the influences of body build, drug metabolizing enzyme genotype, blood sampling time, and moderate recent ethanol intake were investigated.

Methods

The 6‐hydroxychlorzoxazone : chlorzoxazone (metabolic) ratio in plasma was measured at 2 h in 28 male and nine female volunteers following a single oral dose of 500 mg chlorzoxazone. Similarly, the metabolic ratios at 4 h and 6 h were measured in 20 of the males. The metabolic ratio at 2 h was also determined 1.5 and 2.5 years later in 13 and seven males, respectively, and weekly for 3 weeks in seven males, after a dose of 500 mg, once at higher (750 mg) and lower (250 mg) doses, and once (500 mg) following moderate ethanol intake (0.5 g kg−1 body weight) the preceding evening. Genotypes were determined for CYP2E1 as well as for N‐acetyltransferase 2 and glutathione transferase M1.

Results

Excluding an outlier (ratio = 1.6) the metabolic ratio at 2 h ranged from 0.12 to 0.61 (n = 36). A positive correlation with body weight (r = 0.61, P < 0.001) suggested dose‐dependent metabolism of chlorzoxazone. The metabolic ratio decreased with increasing chlorzoxazone dose (P = 0.01), again suggesting dose‐dependent metabolism. Long‐term (yearly intervals) and short‐term (weekly intervals) intra‐ and interindividual variabilities in metabolic ratio were similar (30% and 63%vs 28% and 54%, respectively). Both inter‐ and intra‐individual variabilities tended to decrease with increasing dose of chlorzoxazone. There was no significant influence of moderate ethanol intake the preceding evening, or of CYP2E1 genotype on the metabolic ratio.

Conclusions

The relatively low intra‐individual variability in the metabolism of chlorzoxazone suggests that a single‐sample procedure may suffice to assess CYP2E1 activity in vivo. However, chlorzoxazone metabolism is dose‐dependent at commonly used doses and it is therefore advisable to adjust the dose for body weight. Moderate intake of ethanol the preceding evening did not significantly affect the chlorzoxazone metabolic ratio.
Th1 and Th2 cytokine production is suppressed at the level of transcriptional regulation in Kawasaki disease
Authors: KIMURA J , TAKADA H , NOMURA A , OHNO T , MIZUNO Y , SAITO M , KUSUHARA K , HARA T
Published: July 2004

To clarify the functional state of T cells in Kawasaki disease, we analysed mRNA expression levels of Th1/Th2 cytokines (IFN-γ and IL-4) along with Th1/Th2-inducing transcription factors, T-bet and GATA-3, which play pivotal roles in the development of Th1 and Th2 cells, respectively. By real-time PCR, IFN-γ mRNA levels in peripheral blood mononuclear cells (PBMNC) were significantly decreased in Kawasaki disease patients compared with those with measles, and tended to be lower than those in healthy controls. T-bet mRNA levels were significantly decreased in patients with Kawasaki disease compared with healthy controls. In addition, IL-4 and GATA-3 mRNA levels were significantly decreased in Kawasaki disease compared with healthy controls. Regulatory cytokine mRNA levels (TGF-β and IL-10) were also decreased in Kawasaki disease. The mRNA levels of IFN-γ showed a significant positive correlation with those of T-bet in Kawasaki disease. These results suggest that the suppressed function of Th1 and Th2, associated with the suppression of both T-bet and GATA-3 gene expression, may be one of the immunological characteristics of Kawasaki disease.
Disruption of MAP kinase activation and nuclear factor binding to the IL-12 p40 promoter in HIV-infected myeloid cells
Authors: CHAMBERS K A , PARKS R J , ANGEL J B
Published: July 2004

Progressive immunodeficiency in HIV infection is paralleled by a decrease in IL-12 production, a cytokine crucial for cellular immune function. Here we examine the molecular mechanisms by which HIV infection suppresses IL-12 p40 expression. HIV infection of THP-1 myeloid cells resulted in decreased LPS-induced nuclear factor binding to the NF-κB, AP-1, and Sp1 sites of the IL-12 p40 promoter. By site-directed mutagenesis we determined that each of these sites was necessary for transcriptional activation of the IL-12 p40 promoter. Binding of NF-κB p50, c-Rel, p65, Sp1, Sp3, c-Fos, and c-Jun proteins to their cognate nuclear factor binding sites was somewhat impaired by HV infection, although a role for other as yet unidentified factors cannot be dismissed. The cellular levels of these transcription factors were unaffected by HIV infection, with the exception of a decrease in expression of NF-κB p65, consistent with the observed decrease in its binding to the IL-12 p40 promoter following HIV infection. Analysis of regulation of upstream LPS-induced MAP kinases demonstrated impaired phosphorylation of JNK and p38 MAPK, and suppressed phosphorylation and degradation of IκBα following HIV infection. These results suggest that alterations in nuclear factor binding to numerous sites in the IL-12 p40 promoter, together may contribute to the suppression in IL-12 p40 transcription previously reported. These effects on nuclear factor binding may be a direct effect of HIV infection on the IL-12 p40 promoter, or may occur indirectly as a consequence of altered MAP kinase activation.
The ratio of n-6 to n-3 fatty acids in maternal diet influences the induction of neonatal immunological tolerance to ovalbumin
Authors: KOROTKOVA M , TELEMO E , YAMASHIRO Y , HANSON L Å , STRANDVIK B
Published: July 2004

Prevalence of allergy is increasing in many countries and might be related to changed environmental factors, such as dietary fatty acids (FA). The present study investigates whether dietary ratio of n-6 to n-3 FA influences the induction of immunological tolerance to ovalbumin (OA) in neonatal rats. During late gestation and throughout lactation Sprague-Dawley rats were fed a diet containing 7% linseed oil (n-3 diet), sunflower oil (n-6 diet) or soybean oil (n-6/n-3 diet). At 10–16 days of age the rat offspring were subsequently exposed, or not, to OA via the milk. The offspring were weaned onto the same diets as the mothers and immunized with OA and the bystander antigen human serum albumin (HSA). In the offspring on the n-3 diet exposure to OA via the milk resulted in lower delayed type hypersensitivity reaction (DTH) and antibody responses against both OA and HSA, compared to those in the offspring not exposed to OA, indicating the induction of oral tolerance. In the offspring on the n-6 diet, the exposure to OA led to depressed specific immune responses against only OA, not HSA. In the offspring on the n-6/n-3 diet oral exposure to OA did not influence immune responses against OA, or HSA. The results indicate that the dietary ratio of n-6/n-3 FA is important for the induction of neonatal oral tolerance. Thus nonoptimal feeding may have effects on the development of immunological tolerance to dietary antigen ingested by the mother. The ratio of n-6/n-3 FA in the diet may be considered in the context of increased prevalence of allergy.
Rapid simultaneous measurement of multiple cytokines using 100 µl sample volumes − association with neonatal sepsis
Authors: HODGE G , HODGE S , HASLAM R , MCPHEE A , SEPULVEDA H , MORGAN E , NICHOLSON I , ZOLA H
Published: July 2004

Early diagnosis of neonatal infection has proved problematic due to the inadequacy of currently available laboratory tests. Neonatal sepsis is associated with an increase in plasma-derived cytokine levels, but an increase of a single cytokine cannot identify neonatal sepsis specifically and multiple cytokine levels are required. The time constraints and relatively large volume of plasma required to measure multiple cytokines from newborn infants by conventional enzyme-linked immunosorbent assay (ELISA) techniques is prohibitive. We therefore applied cytometric bead array (CBA) technology for simultaneous measurement of multiple cytokines from a group of 18 term neonates with infection confirmed by culture and a control group. ‘Normal’ ranges were established for each cytokine from 1–7-, 8–14- and 15–21-day-old newborns. There was no significant change in the levels of cytokines from infants in different control age groups, suggesting that basal cytokine levels are unchanged in the first 3 weeks of life. In the patient groups, however, there was a significant difference in several cytokines between the different age groups. Interleukin (IL)-6, IL-10 and IL-12 were increased significantly in the 1–7-day-old patient group compared to either the 8–14 and 15–21 age group, suggesting that infection in utero is associated with increased levels of these cytokines compared to infection acquired following birth. When individual patient cytokine levels were compared to normal control reference ranges, two patients failed to show significant elevation of any cytokine tested. All other patients showed elevated levels of between one and nine cytokines tested (mean of 4·6). There was no correlation between elevated cytokine levels and types of infective organism or patient age. In conclusion, neonatal sepsis is associated with the elevation of multiple plasma cytokines. The use of CBA kits is a rapid, easy, low sample volume and sensitive method to measure multiple plasma cytokines.
Effect of prolactin on carcinoembryonic antigen-specific cytotoxic T lymphocyte response induced by dendritic cells
Authors: MATERA L , BELTRAMO E , MARTINUZZI E , BUTTIGLIERI S
Published: July 2004

The cytokine hormone prolactin (PRL) has been shown previously to modulate native cellular responses and maturation of antigen-presenting cells. Here we have addressed its effect on the antigen-specific response of cytotoxic T lymphocytes (CTL). CTL were generated from HLA-A2 lymphocytes after three rounds of stimulation with autologous dendritic cells loaded with HLA-A2-restricted carcinoembrionic antigen (CEA) Cap-1 (YLSGANLNL) peptide. Selected cultures were expanded on cytokine-supplemented feeder-layers, enriched for CD8+ lymphocytes and analysed for PRL-receptor (PRL-R) expression and PRL responsiveness. Resting CD8+ lymphocytes were negative for PRL-R, whereas antigen-activated CD8+ lymphocytes derived from long-term cultures were highly positive. Results of a 51Cr release assay showed CTL killing of CEA-loaded, but not unloaded, T2 cell line and the CEA-positive gastric carcinoma cell line KATO, but not of the CEA-negative T leukaemia cell line Jurkat. Interferon (IFN)-γ release, evaluated in an ELISPOT assay against CEA-loaded T2, was enhanced (P < 0·05) by concentrations of PRL (12–25 ng/ml) very close to the physiological levels (6–20 ng/ml), but was decreased (P < 0·05) by high concentrations (200 ng/ml). Pre-incubation of the stimulators with the anti-MHC class I MoAb W6·32 induced a 40–60% decrease of the PRL-boosted IFN-γ release, thus proving the MHC restriction of the lymphocyte response. Cytotoxicity against CEA-loaded T2 and KATO cell lines was also increased by 12–25 ng (P < 0·05) and decreased (P < 0·05) by 200 ng PRL. Pre-incubation of CTL with an antibody specific for the PRL-R almost completely abrogated this effect.
Mycoplasma infection induces a scleroderma-like centrosome autoantibody response in mice
Authors: GAVANESCU I , PIHAN G , HALILOVIC E , SZOMOLANYI-TSUDA E , WELSH R M , DOXSEY S
Published: July 2004

Development of autoantibodies to intracellular molecules is a universal feature of autoimmune diseases and parallels onset of chronic inflammatory pathology. Initiating antigens of disease-specific autoantibody responses are unknown. We previously showed that the major targets of autoantibodies in scleroderma are centrosomes, organelles involved in mitotic spindle organization. Here we show that centrosome autoantibodies are induced in mice by mycoplasma infection. The centrosome-specific antibody response involves class switching of preexisting IgM to IgG isotypes, suggesting a T cell-dependent mechanism. The antibody response spreads to include additional intracellular targets, with newly recruited autoantibody specificities arising as IgM isotypes. Antibiotic treatment of mice prevents autoantibody development. Centrosome autoantibodies may provide an aetiological link between infection and human autoimmunity and suggest novel therapeutic strategies in these disorders.
The number of CD8+ T cells and NKT cells increases in the aqueous humor of patients with Behçet's uveitis
Authors: YU H G , LEE D S , SEO J M , AHN J K , YU Y S , LEE W J , CHUNG H
Published: July 2004

To determine whether there are differences in the immunopathogenesis of different endogenous uveitis syndromes, the phenotypic characteristics of immune cells were analysed among patients with endogenous uveitis. The aetiology of the uveitis included idiopathic recurrent acute anterior uveitis (18 patients), idiopathic intermediate uveitis (13 patients), Behçet's uveitis (17 patients), Vogt–Koyanagi–Harada syndrome (7 patients), and so on. Flow cytometric analysis was performed using immune cells of the aqueous humor and the peripheral blood during the active phase of intraocular inflammation, and monoclonal antibodies to CD3, CD4, CD8, CD14, CD19, CD56, TCR γδ, pan TCR αβ and Vα24. CD8+ T cells were predominant in the aqueous humor of the patients with Behçet's uveitis, whereas CD4+ T cells were mainly found in the aqueous humor of patients other than those with Behçet's uveitis. The number of NKT (CD3+CD56+) cells was significantly higher both in the aqueous humor and the peripheral blood of the patients with Behçet's uveitis compared with the other groups (P < 0·05). CD8+CD56+ cells were the predominant subtype of the increased NKT cells in patients with Behçet's uveitis. In addition, intraocular infiltration of CD14+ cells significantly differed among the uveitis patients (P < 0·05). These results suggest that the immunopathogenesis of endogenous uveitis can vary between syndromes, and that CD8+CD56+ NKT cells may play an important role in the immunopathogenesis of Behçet's uveitis.
Increase in Ksp37-positive peripheral blood lymphocytes in mild extrinsic asthma
Authors: KUEPPER M , KOESTER K , BRATKE K , MYRTEK D , OGAWA K , NAGATA K , VIRCHOW J C , LUTTMANN W
Published: July 2004

Killer-specific secretory protein of 37 kDa (Ksp37), identified as a Th1/Tc1 specific secretory protein is expressed preferentially in cytotoxic T lymphocytes (CTL) and natural killer (NK) cells and might be involved in essential processes of CTL-mediated immunity. Although extrinsic asthma is linked currently to a Th2-dominated pathogenesis, there is increasing evidence for Th1/Tc1-mediated processes in the aetiopathology of asthma. CTL from patients with asthma have been shown to express cytokines and effector molecules which were different from healthy controls. We hypothesized that Ksp37 could indicate the involvement of CTL in the pathogenesis of extrinsic asthma. We therefore investigated Ksp37 expression in PBMC from patients with mild extrinsic asthma (n = 7) and healthy controls (n = 7). Flow cytometric analysis was used to quantify Ksp37+ cells and to investigate cellular Ksp37 expression as relative mean fluorescence intensities (MFI). We found a significantly (P = 0·016) higher percentage of Ksp37+ cells within the total lymphocyte population obtained from patients with mild extrinsic asthma compared with healthy controls. Subdifferentiation revealed a significant difference limited exclusively to the CD8+ subset (P = 0·010). In addition, Ksp37 secretion from cultured peripheral blood mononuclear cells (PBMC) and MFI of Ksp37+ lymphocytes were increased in patients with asthma compared with healthy controls. We conclude that mild extrinsic asthma appears to be associated with an increased expression of the Tc1 related protein Ksp37. The functional role of Ksp37 in the pathogenesis of asthma remains to be elucidated.
Anti-actin IgA antibodies in severe coeliac disease
Authors: GRANITO A , MURATORI P , CASSANI F , PAPPAS G , MURATORI L , AGOSTINELLI D , VERONESI L , BORTOLOTTI R , PETROLINI N , BIANCHI F B , VOLTA U
Published: July 2004

Anti-actin IgA antibodies have been found in sera of coeliacs. Our aim was to define the prevalence and clinical significance of anti-actin IgA in coeliacs before and after gluten withdrawal. One hundred and two biopsy-proven coeliacs, 95 disease controls and 50 blood donors were studied. Anti-actin IgA were evaluated by different methods: (a) antimicrofilament positivity on HEp-2 cells and on cultured fibroblasts by immunofluorescence; (b) anti-actin positivity by enzyme-linked immuosorbent assay (ELISA); and (c) presence of the tubular/glomerular pattern of anti-smooth muscle antibodies on rat kidney sections by immunofluorescence. Antimicrofilament IgA were present in 27% of coeliacs and in none of the controls. Antimicrofilament antibodies were found in 25 of 54 (46%) coeliacs with severe villous atrophy and in three of 48 (6%) with mild damage (P < 0·0001). In the 20 patients tested, antimicrofilaments IgA disappeared after gluten withdrawal in accordance with histological recovery. Our study shows a significant correlation between antimicrofilament IgA and the severity of intestinal damage in untreated coeliacs. The disappearance of antimicrofilament IgA after gluten withdrawal predicts the normalization of intestinal mucosa and could be considered a useful tool in the follow-up of severe coeliac disease.
Impaired synthesis and action of antiaggregating cyclic nucleotides in platelets from obese subjects: possible role in platelet hyperactivation in obesity
Authors: Anfossi G. , Russo I. , Massucco P. , Mattiello L. , Doronzo G. , De Salve A. , Trovati M.
Published: July 2004

Background  Subjects with central obesity exhibit platelet hyperactivity, which is involved in the atherosclerotic process and therefore can account for the increased risk of cardiovascular morbidity and mortality. The aim of the study was to evaluate whether alterations of platelet function in obesity involve synthesis and/or action of the two antiaggregating cyclic nucleotides adenosine 3′,5′‐cyclic monophosphate (cAMP) and guanosine 3′,5′‐cyclic monophosphate (cGMP).

Materials and methods  In platelets from 16 obese and 15 control subjects we investigated the influence on platelet responses to the Adenosine‐5‐diphosphate sodium salt (ADP) exerted by (i) prostacyclin analogue Iloprost (0·31–5 nmol L−1) and the cAMP analogue 8‐bromo‐cAMP (10–500 µmol L−1); and by (ii) nitric oxide (NO) donor sodium nitroprusside (SNP) (5–100 µmol L−1) and the cGMP analogue 8‐bromo‐cGMP (10–500 µmol L−1). IC50 (minimal concentration of each inhibitor necessary to reduce platelet response to ADP by half) was determined. Iloprost and SNP ability to increase cyclic nucleotides was also measured.

Results  Significantly greater IC50 were observed in obese subjects than in healthy controls (1·59 ± 0·16 vs. 0·80 ± 0·08 nmol L−1, P = 0·0001 for Iloprost, and 27·6 ± 6·5 vs. 7·0 ± 1·7 µmol L−1, P = 0·006, for SNP); when data from control and obese subjects were pooled together, IC50 of Iloprost and SNP correlated with the homeostasis model assessment (HOMA IR), which is a parameter used to measure the insulin resistance (r = 0·588, P = 0·029 and r = 0·640, P = 0·006, respectively). Also the antiaggregating effect of 8‐Br‐cAMP and 8‐Br‐cGMP was smaller in the obese subjects. Finally, the ability of Iloprost to increase platelet cAMP and the ability of SNP to increase both cGMP and cAMP were reduced in obese subjects.

Conclusions  Platelet resistance to the antiaggregating effects of prostacyclin and NO in obesity is attributable to impairment of cyclic nucleotide synthesis and action. As cyclic nucleotides are the main effectors of platelet antiaggregation, the resistance to them can account for platelet hyperactivity in obesity.
Cytokine gene polymorphisms and breast cancer susceptibility and prognosis*
Authors: Smith K. C. , Bateman A. C. , Fussell H. M. , Howell W. M.
Published: July 2004

Single nucleotide polymorphisms (SNPs) in the promoter regions of cytokine genes are associated with differential levels of cytokine expression. We hypothesized that these SNPs might influence breast tumour development and progression by affecting the efficiency of the antitumour immune response and/or pathways of angiogenesis. A total of 144 female breast cancer patients and 263 cancer‐free population controls were genotyped for the interleukin (IL)‐1β−511 (T/C), IL‐6 −174 (G/C), tumour necrosis factor (TNF)‐α−308 (A/G), IL‐10 −1082 (A/G), IL‐8 −251 (A/T) and vascular endothelial growth factor (VEGF) −1154 (A/G) SNPs, using amplification refractory mutation system polymerase chain reaction (ARMS‐PCR) and TaqMan® (Applied Biosystems, Foster City, CA, USA) 5′ nuclease assays for allelic discrimination. No significant associations were seen. Patient–control comparisons revealed a non‐significant trend for association between the TNF‐α−308 GG genotype and breast cancer compared to controls (79.7 vs. 68.2%, P = 0.03, Pc = 0.54). Stratification of the patient group according to the Nottingham Prognostic Index and individual prognostic factors revealed trends for association between IL‐6 −174 GC and IL‐8 −251 AA genotypes and markers of poor prognosis (P = 0.04, Pc = 0.72 and P = 0.02, Pc = 0.36, respectively). There were also trends for associations between VEGF −1154 AG and IL‐1β−511 TC genotypes and markers of good prognosis (P = 0.02, Pc = 0.36 and P = 0.05, Pc = 0.90, respectively). These results suggest that the role of cytokine promoter SNPs in both susceptibility to and prognosis in breast cancer requires further investigation in a larger study.
High nucleotide and amino acid sequence similarities in tumour necrosis factor‐α amongst Indian buffalo (Bubalus bubalis), Indian cattle (Bos indicus) and other ruminants*
Authors: Gupta P. K. , Bind R. B. , Walunj S. S. , Saini M.
Published: July 2004

Tumour necrosis factor‐α (TNF‐α) mRNA from Indian water buffalo (Bubalus bubalis) and Indian cattle (Bos indicus) was reverse transcribed and amplified using reverse transcriptase–polymerase chain reaction (RT‐PCR). The nucleotide sequences of cDNAs were determined after cloning into pGEM‐T‐Easy vector (Promega, Madison, WI) and compared with reported nucleotide sequences of TNF‐α cDNA from other species. The nucleotide sequences of TNF‐α from Indian cattle revealed significantly high similarities at nucleotide (99.2%) and amino acid (100%) levels with those of cattle (Bos taurus; Zebu). The sequences from buffalo had 98.4% nucleotide and 99.1% amino acid similarities with Indian cattle, indicating functional cross‐reactivity. One amino acid deletion at position 63 and one substitution (A→P) at position 64 were observed in buffalo compared with Indian cattle. The amino acid deletion at position 63 was predicted due to differences in pre‐mRNA splicing.
Metapopulation structure of stream‐dwelling Dolly Varden charr inferred from patterns of occurrence in the Sorachi River basin, Hokkaido, Japan
Authors: Koizumi I. , Maekawa K.
Published: July 2004

1. We investigated the occurrence (presence or absence) of stream‐dwelling Dolly Varden (Salmonidae:Salvelinus malma) in the 78 tributaries of the Sorachi River system, Hokkaido, Japan to examine evidence for metapopulation structure.

2. We hypothesised (i) that the occurrence of Dolly Varden would be less likely in small tributaries because of the higher extinction risk and (ii) that occurrence would be less likely in isolated tributaries because of reduced rates of dispersal and colonisation. In addition to these two general predictions, we hypothesised (iii) that the occurrence would be more likely in spring‐fed tributaries, because Dolly Varden prefer cold water as present in these systems.

3. When all the tributaries were included in a multiple logistic regression analysis, habitat size and habitat type (spring‐fed versus non‐spring‐fed tributaries) were selected for the best model, whereas isolation was not important. However, when tributaries in the lower Sorachi basin (an area likely influenced by human activities) were excluded from the analysis, the effect of isolation became significant.

4. Our results indicate there is a potential metapopulation structure of the Dolly Varden in the Sorachi River basin. Furthermore, it is possible that metapopulation dynamics follow a clinal pattern. In the upper and middle reaches of the Sorachi River basin, an equilibrium between extinction and colonisation of Dolly Varden populations is a possibility. In the lower portion of the basin, isolation appeared to be less important, and extinction rate may be driving patterns of occurrence toward systematic extinction of local populations.

5. Our results illustrate the possibility of spatial variation in the processes influencing occurrence of Dolly Varden within a single river basin, and the potential influences of habitat alteration on extinction and colonisation processes.
Metabolism of dissolved organic carbon by planktonic bacteria and mixotrophic algae in lake neutralisation experiments
Authors: Tittel Jörg , Kamjunke Norbert
Published: July 2004

1. Lakes formed in mining pits often contain high concentrations of dissolved ferric iron and sulphate (e.g. 2 and 16 mmol L−1, respectively) and the pH is buffered between 2.5 and 3.5. Efforts to neutralise their water are based on the stimulation of lake internal, bacterial iron‐ and sulphate reduction. Electron donors may be supplied by organic carbon compounds or indirectly by enhancement of primary production. Here, we investigated the function of mixotrophic algae, which can potentially supplement or deplete the organic carbon pool, in the carbon metabolism and alkalinity budget of an acidic mining lake.

2. Two weeks after organic substrates had been added in a large in situ mesocosm of 30 m diameter, a bloom of Chlamydomonas occurred, reaching a biovolume of 80 mm3 L−1. Growth experiments using filtered lake water showed that the alga reduced the overall dissolved organic carbon (DOC) concentration despite significant photosynthetic activity. However, when Chlamydomonas were grown together with natural bacterioplankton, net DOC consumption did not increase.

3. Uptake experiments using [14C]‐glucose indicated that bacteria dominated glucose uptake and remineralisation. Therefore, the DOC leached in the water column was processed mainly by planktonic bacteria. Leached DOC must be regarded as loss, not transferred by larger organisms to the sediment, where reduction processes take place.

4. From phytoplankton biomass and production 2 years after fertilisation we estimated that pelagic photosynthesis does not supply an electron donor capacity capable of reducing more than 2% of actual stock of acidity per year. We estimated that only the benthic primary production was in a range to compensate for ongoing inputs of iron and sulphate.
Assessing the short‐term response of stream diatoms to acidity using inter‐basin transplantations and chemical diffusing substrates
Authors: Hirst Heike , Chaud Fabrice , Delabie Clotilde , Jüttner Ingrid , Ormerod S. J.
Published: July 2004

1. Acid‐base status has major effects on diatoms, but there is little information on their short‐term response to changing acidity. This is despite the use of diatoms as bioindicators in streams where acid episodes are important during rainstorms (hours to days) or snowmelt (days to weeks). In the Llyn Brianne experimental catchments (Wales, UK), we attempted to mimic the effects of short‐term acidification by (i) reciprocally transplanting diatoms between two streams of contrasting acidity and (ii) using acid‐diffusing substrates.

2. Diatom diversity decreased rapidly on substrata transplanted from the circumneutral into the acidic stream, and increased in the reciprocal transplantation. Changes in dominant taxa occurred within three days in the acidic stream because of the rapid growth ofEunotia exigua, and by nine days in the circumneutral stream because of the proliferation ofAchnanthidium minutissimum. Transplants were near indistinguishable from ambient assemblages by day 12.

3. There were no effects of enclosures on assemblage composition, but diatoms responded more rapidly to altered chemistry in enclosures with coarse mesh (26 × 50 mm) than finer mesh (320 μm).

4. Chemical diffusing substrates comprised terracotta tiles attached to dosing reservoirs that created locally acid (using H2SO4) or metal‐rich conditions (using MnSO4) in the circumneutral stream over 26 days. Diatom responses were compared with reference substrates dosed with deionised or circumneutral stream water, and we also assessed whether effects were moderated by macroinvertebrate grazers.

5. Surface pH was lower by 1–2 pH units on acid‐dosed substrates than on reference tiles or in surrounding streamwater. Grazed assemblages on acid‐dosed substrates differed significantly from ungrazed reference assemblages, acquiring significantly greater relative abundance ofEunotiaspp. However, the magnitude of response was less than in the between‐stream transplantations either because (i) metal exposure and base cation concentrations differed between the transplants and dosing substrates or (ii) diatom response to reduced pH on the diffusing substrates was restricted by the scarcity of acidobiontic diatoms in the circumneutral stream. Similar filter, founder or dominance effects might also affect diatom responses to real acid episodes.

6. These data show that diatom assemblages can respond rapidly and directly to changes in acid‐base status, but short‐term acidification might affect diatoms more rapidly than subsequent recovery. Because the experimental methods used were imperfect representations of episodic effects, diatom response to real acid events requires further field evaluation.
Movements and habitat use by PIT‐tagged Atlantic salmon parr in early winter: the influence of anchor ice
Authors: Roussel Jean‐Marc , Cunjak Richard A. , Newbury Robert , Caissie Daniel , Haro Alexander
Published: July 2004

1. Movements and habitat use by Atlantic salmon parr in Catamaran Brook, New Brunswick, were studied using Passive Integrated Transponder technology. The fish were tagged in the summer of 1999, and a portable reading system was used to collect data on individual positions within a riffle‐pool sequence in the early winter of 1999. Two major freezing events occurred on November 11–12 (Ice 1) and November 18–19 (Ice 2) that generated significant accumulations of anchor ice in the riffle.

2. Individually tagged parr (fork length 8.4–12.6 cm,n = 15) were tracked from 8 to 24 November 1999. Over this period, emigration (40%) was higher from the pool than from the riffle. Of the nine parr that were consistently located, seven parr moved <5 m up‐ or downstream, and two parr moved more than 10 m (maximum 23 m). Parr moved significantly more by night than by day, and diel habitat shifts were more pronounced in the pool with some of the fish moving closer to the bank at night.

3. DuringIce 2, there was relatively little movement by most of the parr in the riffle beneath anchor ice up to 10 cm in thickness. Water temperature was 0.16 °C above the freezing point beneath anchor ice, suggesting the existence of suitable habitats where salmon parr can avoid supercooling conditions and where they can have access to low velocity shelters. To our knowledge, these are the first data on habitat use by Atlantic salmon parr under anchor ice.
Selective feeding of Arctodiaptomus salinus (Copepoda, Calanoida) on co‐occurring sibling rotifer species
Authors: Lapesa Sara , Snell Terry W. , Fields David M. , Serra Manuel
Published: July 2004

1. Using two‐ and three‐dimensional video recordings, we examined the steps involved in predation that lead to the differential vulnerability of three sympatric rotifer sibling species (Brachionus plicatilis, B. ibericus and B. rotundiformis) to a co‐occurring, predatory, calanoid copepod (Arctodiaptomus salinus).

2. Brachionus rotundiformis, the smallest prey tested, was the most vulnerable with the highest encounter rate, probability of attack, capture and ingestion, and the lowest handling time.

3. Comparison of our results with those of a previous study shows that A. salinus is a more efficient predator than a co‐occurring cyclopoid copepod (Diacyclops bicuspidatus odessanus) feeding on these same rotifer species. However, despite its higher capture rates, A. salinus seems to be less selective than D. b. odessanus based on attack distances and prey handling times.

4. The differential vulnerability to both calanoid and cyclopoid copepod predation can help explain the coexistence and seasonal succession of these co‐occurring rotifer species.